tretinoin and Uterine-Cervical-Neoplasms

Retinoids are natural and synthetic derivatives of vitamin A, which can be obtained from animal products (milk, liver, beef, fish oils, and eggs) and vegetables (carrots, mangos, sweet potatoes, and spinach). Retinoids regulate various important cellular functions in the body through specific nuclear retinoic-acid receptors and retinoid-X receptors, which are encoded by separate genes. Retinoic-acid receptors specifically bind tretinoin and alitretinoin, whereas retinoid-X receptors bind only alitretinoin. Retinoids have long been established as crucial for several essential life processes-healthy growth, vision, maintenance of tissues, reproduction, metabolism, tissue differentiation (normal, premalignant cells, and malignant cells), haemopoiesis, bone development, spermatogenesis, embryogenesis, and overall survival. Therefore, deficiency of vitamin A can lead to various unwanted biological effects. Several experimental and epidemiological studies have shown the antiproliferative activity of retinoids and their potential use in cancer treatment and chemoprevention. Emerging clinical trials have shown the chemotherapeutic and chemopreventive potential of retinoids in cancerous and precancerous conditions of the uterine cervix. In this review, we explore the potential chemopreventive and therapeutic roles of retinoids in preinvasive and invasive cervical neoplasia.

Topics: Antineoplastic Agents; Female; Gene Expression Regulation, Neoplastic; Humans; Papillomavirus Infections; Receptors, Retinoic Acid; Retinoid X Receptors; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Invasive cervical cancer accounts for 11.6% of all cancers worldwide and is the second most common cancer among women. It is the most common cancer among women living in less developed countries. Although infection with oncogenic-type human papillomaviruses (HPV) is associated with most cases of cervical cancer, HPV infection alone is an insufficient cause of cervical cancer. Research from the last two decades suggests a role for nutrients in the prevention of cervical cancer. However, results from phase III folic acid and beta-carotene chemoprevention trials have been negative. Potential reasons for the lack of treatment effect are discussed within the context of cervical carcinogenesis.

Topics: beta Carotene; Dietary Supplements; Female; Folic Acid; Humans; Treatment Outcome; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

We review the current status of prevention trials in the management of cervical intraepithelial neoplasia (CIN) and cervical cancer. Two large, randomized controlled trials have shown that folic acid is inactive in reversing low to moderate grade CIN. A large randomized trial of locally applied beta-trans retinoic acid showed that the agent was effective in reversing moderate but not severe CIN. Results from a pilot trial involving 30 patients with CIN I (mild dysplasia) and CIN II (moderate dysplasia) indicate that beta-carotene can suppress CIN; a large ongoing randomized trial will answer the question more definitively.

Topics: Antioxidants; beta Carotene; Carotenoids; Female; Humans; Randomized Controlled Trials as Topic; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Retinoic acid and interferon-alpha have limited single-agent activity in advanced cancer. Cell culture data indicate that in combination these agents have enhanced activity (modulating growth and differentiation) in a number of malignant cell types. Recent clinical work in advanced squamous cell carcinoma reports major activity with this regimen. This paper reviews the preclinical and clinical data testing retinoic acid in combination with interferons and presents recent work integrating these agents with radiotherapy in locally advanced cervical cancer.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Combined Modality Therapy; Female; Humans; Interferon-alpha; Middle Aged; Skin Neoplasms; Tretinoin; Uterine Cervical Neoplasms

Because a combination of retinoic acid, interferon-alpha, and radiation therapy demonstrated synergistic action and effectiveness to treat advanced cervical cancers in earlier studies, we designed this randomized phase 2 open-label trial to assess efficacy and safety of interferon alpha-2b (IFN) and 13-cis-retinoic acid (RA) administered concomitantly with radiation therapy (IFN-RA-radiation) to treat stage III cervical cancer.. Stage III cervical cancer patients were randomized to study and control groups in a 1:1 ratio. All patients were treated with radiation therapy; study arm patients received IFN (3 × 10(6) IU subcutaneously) 3 times a week for 4 weeks and daily RA (40 mg orally) for 30 days starting on day 1 of radiation, whereas control arm patients received weekly cisplatinum (40 mg/m(2)) for 5 weeks during radiation. Patients were followed for 3 years. The primary endpoint was overall survival at 3 years.. Patients in the study (n=104) and control (n=105) groups were comparable for clinicopathological characteristics, radiation therapy-related variables and treatment response. Proportions of disease-free patients in the study and control groups were 38.5% and 44.8%, respectively, after median follow-up of 29.2 months. Hazard ratios were 0.67 (95% confidence interval [CI]: 0.44-1.01) and 0.69 (95% CI: 0.44-1.06) for overall and disease-fee survival, respectively, comparing the study group to control, and demonstrated an inferior outcome with RA-IFN-radiation, although differences were statistically nonsignificant. Kaplan-Meier curves of disease-free and overall survival probabilities also showed inferior survival in the study group compared to those in the control. Acute toxicities of chemoradiation were significantly higher with 2 acute toxicity-related deaths.. Treatment with RA-IFN-radiation did not demonstrate survival advantage over chemoradiation despite being less toxic. The trends predicted an inferior outcome with the RA-IFN combination.

Topics: Aged; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Brachytherapy; Chemoradiotherapy; Cisplatin; Disease-Free Survival; Drug Administration Schedule; Female; Humans; Interferon alpha-2; Interferon-alpha; Kaplan-Meier Estimate; Middle Aged; Prospective Studies; Radiotherapy Dosage; Recombinant Proteins; Tretinoin; Uterine Cervical Neoplasms

We wanted to examine the tolerability and the toxicity of the chemoimmunotherapy in combination with radiotherapy in patients with stage III B cervical cancer. Sixteen patients were selected for examination and the experience of the Gynecological Cancer Center in Sidney was summarized as well. We used cisplatin in dose from 10 to 20 mg/m2 intravenously weekly, 5-Fluoruracil/5 Fu/500 mg/m2--24 hours infusion, weekly, interferon-alpha--2a 3MU subcutaneously for 3 days course weekly and 13 cis-retinoic acid 0.5 mg/kg daily by month together with 8 weeks period of radiotherapy after the end of the radiotherapy these medicaments were given for another 4 weeks. According to the dose of the cisplatin medicaments we divided the patients into 3 groups--first group--6 patients with dose to 10 mg/m2 cisplatin, second group--6 patients to 15 mg/m2 cisplatin, third group 4 patients with dose to 20 mg/m2 cisplatin. The patients from the first and second group tolerated the cisplatin. In the third group--4 patients had leucopenia. After the end of chemoimmunotherapy and the patients recovered from the toxicity, they prolonged with radiotherapy. In conclusion from our small experience the cisplatin--15 mg/m2, 5-Fu-500 mg/m2 weekly, interferon alpha--2a 3MU subcutaneously for 3 days course weekly and 13-cis-retinoic acid 0.5 mg/kg daily in combination with radiotherapy for stage III B cervical cancer we can say that and the dose is tolerated extremely well. The leucopenia is the dose-limiting toxicity for this group of patients. The survival rate will be followed. The first results show better survival rate for this group of patients.

Topics: Antimetabolites, Antineoplastic; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Cisplatin; Combined Modality Therapy; Drug Administration Schedule; Female; Fluorouracil; Humans; Infusions, Intravenous; Injections, Subcutaneous; Interferon alpha-2; Interferon-alpha; Neoplasm Staging; Recombinant Proteins; Tretinoin; Uterine Cervical Neoplasms

The objective of this study was to determine an effective dose for all-trans retinoic acid (atRA) delivered with a cervical cap and sponge for 4 days to women with cervical intraepithelial neoplasia (CIN) II/III.. Study participants made up of 175 women with biopsy-proven CIN II/III were randomized to four consecutive days of atRA at one of three doses (0.16%, 0.28%, and 0.36%) or placebo. All subjects underwent a repeat colposcopy evaluation and biopsy of the cervix at 12 weeks.. The study participants mean ages were 27.6 years. The racial distribution was 63% Caucasian, 27% African American, and 8% other. Among participants, 93% were human papillomavirus-positive at baseline with 68% positive for high-risk types. The disease response at 12 weeks to atRA or placebo was not significantly different (P = 0.49) among the four dose groups. Participants with CIN II at baseline were more likely to be free of disease at 12 weeks than participants with CIN III at baseline (P = 0.003). There were no reported systemic adverse events related to drug or placebo exposure and only mild local self-reported and clinician-detected toxicities.. Lower concentrations of atRA applied with a cervical cap for 4 days were no more effective than placebo. However, the rate of histologic regression in biopsied CIN II/III patients was high even over a short time interval, and emphasizes the importance of having a placebo arm and an adequate sample size.

Topics: Adolescent; Adult; Antineoplastic Agents; Chemoprevention; Colposcopy; Contraceptive Devices, Female; Dose-Response Relationship, Drug; Female; Humans; Middle Aged; Placebos; Risk Factors; Sample Size; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

9-Cis-retinoic acid (aliretinoin) is a pan-retinoid receptor agonist and has been demonstrated in preclinical models to have potent chemoprevention effects. The purpose of this study was to determine the utility of using aliretinoin as a chemoprevention agent in cervical dysplasia. Patients with histological evidence of cervical intraepithelial neoplasia (CIN) 2/3 were randomized in a double-blind manner to receive high-dose aliretinoin (50 mg), low-dose of aliretinoin (25 mg), or placebo daily for 12 weeks. Compliance and side effects were monitored at various time points during therapy. At the completion of therapy, all of the patients underwent a loop procedure. Histology of pretreatment biopsies was compared with that of loop specimens. One-hundred and fourteen patients with CIN 2/3 were enrolled in the study. In the 112 patients evaluable for toxicity, headache was the most common clinical side effect and was experienced more frequently (74%) in the high-dose aliretinoin group. Eight patients withdrew from the study before completion of study medication because of unacceptable side effects. In the 104 patients evaluable for efficacy, there was no statistical difference in the rate of regression among the placebo (32%), the low-dose aliretinoin (32%), and the high-dose aliretinoin (36%) groups. (P = not significant; power 0.06). Aliretinoin at these dosages and this schedule does not appear to result in significant regression rates in CIN 2/3 patients when compared with placebo. Headache is encountered frequently and may thwart efforts to increase the dose or duration of aliretinoin in future cervical cancer chemoprevention studies. The rate of histological regression in biopsied CIN 2/3 patients is high even over a short time interval, and emphasizes the importance of having a placebo arm and an adequate sample size in cervical dysplasia chemoprevention studies.

Topics: Adolescent; Adult; Alabama; Alitretinoin; Antineoplastic Agents; Biomarkers; Chemoprevention; Cholesterol, HDL; Dose-Response Relationship, Drug; Double-Blind Method; Drug Evaluation; Female; Hemoglobins; Humans; Patient Compliance; Severity of Illness Index; Treatment Outcome; Tretinoin; Triglycerides; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms; Women's Health

Preclinical and clinical data support the study of retinoids and interferon-alpha (IFN-alpha) in advanced squamous cell carcinoma of the uterine cervix (SCC). This phase II randomized trial of the Southwest Oncology Group sought to estimate the response rate for IFN-alpha plus either 13-cis-retinoic acid (13cRA) or all-trans-retinoic acid (ATRA) in women with recurrent cervical SCC.. Eligibility for this trial required bidimensionally measurable locally recurrent or metastatic squamous or adenosquamous carcinoma of the uterine cervix; SWOG performance status of

Topics: Adult; Aged; Carcinoma, Squamous Cell; Female; Humans; Interferon-alpha; Isotretinoin; Middle Aged; Neoplasm Recurrence, Local; Tretinoin; Uterine Cervical Neoplasms

Dysplasia of the uterine cervix is a recognized precancerous condition. Because of the observed ability of retinoids to suppress various cell lines in vitro, a number of clinical studies have examined the effect these agents have on cervical dysplasia, with the object of developing a means of chemoprevention of cervical malignancies in women at risk. Three cervical cancer chemoprevention trials with Retinamide II (RII) have been conducted at the Cancer Institute, Chinese Academy of Medical Sciences, Beijing, China. A pilot study used RII to intervene in cases of precancerous cervical dysplasia. Twenty-seven women with mild, moderate, or severe cervical dysplasia, pathologically confirmed, were treated by RII suppositories, 10 mg QD, given intravaginally for 6 months (each course lasting 3 months). The results indicated that after the second course, the overall response rate was 96.29% and the complete response rate was 88.89%. In general, side effects were mild. A little cervical and vaginal irritation was well tolerated. In the second double-blind study, patients with precancerous cervical lesions were randomized into two groups, one treated with RII suppository intravaginally and the other with a placebo, once daily for 50 days in two courses. Precancerous lesions in 68.76% of patients in the treatment arm disappeared, with an overall effective rate of 74.29% after two courses of treatment with RII. Its curative effect was approximately that of laser beam radiation and electrocautery (P > 0.05), and differed significantly (P < 0.01) from that of traditional antiinflammatories. RII can be a major measure in prevention and treatment of cervical cancer in high-incidence areas in China. In the third trial, we are conducting a randomized double-blind study placebo controlled, in a high-incidence area of cervical cancer (Xiang-Yuan county, Shang Xi Province, China). At present, the patients are being followed up and the study will be completed after 2 years.

Topics: Antineoplastic Agents; China; Double-Blind Method; Female; Humans; Incidence; Pilot Projects; Placebos; Precancerous Conditions; Tretinoin; Uterine Cervical Neoplasms

Locally advanced cervical cancer could be treated with Interferon-alpha plus retinoic acid concomitant with standard radiotherapy. It showed some response in local control. The pattern of relapse and the survival of the patients should be observed in the follow-up period for the conclusion.

Topics: Adult; Aged; Antineoplastic Combined Chemotherapy Protocols; Case-Control Studies; Chemotherapy, Adjuvant; Female; Follow-Up Studies; Humans; Interferon-alpha; Longitudinal Studies; Middle Aged; Treatment Outcome; Tretinoin; Uterine Cervical Neoplasms

The patients with cervical precancerous lesions were double-blindly randomized into two groups. The one was treated with retinamide II (RII) suppository intravaginally and the other with placebo, once daily for 50 days as a course. The results showed precancerous lesions in 68.57% of the patients disappeared, with an overall effective rate of 74.29% after two-course treatment with RII. Its long-term curative effect approximated to that with laser beam radiation and electrocautery (P > 0.05), and differed significantly (P < 0.01) from that with common antiphlogistic. So, RII can be used as a major measure in prevention and treatment for cervical cancer in high-incidence areas in our country.

Topics: Administration, Intravaginal; Double-Blind Method; Female; Follow-Up Studies; Humans; Suppositories; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Retinoids enhance differentiation of most epithelial tissues. Epidemiologic studies have shown an inverse relationship between dietary intake or serum levels of vitamin A and the development of cervical dysplasia and/or cervical cancer. Pilot and phase I investigations demonstrated the feasibility of the local delivery of all-trans-retinoic acid (RA) to the cervix using a collagen sponge insert and cervical cap. A phase II trial produced a clinical complete response rate of 50%.. This randomized phase III trial was designed to determine whether topically applied RA reversed moderate cervical intraepithelial neoplasia (CIN) II or severe CIN.. Analyses were based on 301 women with CIN (moderate dysplasia, 151 women; severe dysplasia, 150 women), evaluated by serial colposcopy, Papanicolaou cytology, and cervical biopsy. Cervical caps with sponges containing either 1.0 mL of 0.372% beta-trans-RA or a placebo were inserted daily for 4 days when women entered the trial, and for 2 days at months 3 and 6. Patients receiving treatment and those receiving placebo were similar with respect to age, ethnicity, birth-control methods, histologic features of the endocervical biopsy specimen and koilocytotic atypia, and percentage of involvement of the cervix at study. Treatment effects were compared using Fisher's exact test and logistic regression methods. Side effects were recorded, and differences were compared using Fisher's exact test.. RA increased the complete histologic regression rate of CIN II from 27% in the placebo group to 43% in the retinoic acid treatment group (P = .041). No treatment difference between the two arms was evident in the severe dysplasia group. More vaginal and vulvar side effects were seen in the patients receiving RA, but these effects were mild and reversible.. A short course of locally applied RA can reverse CIN II, but not more advanced dysplasia, with acceptable local side effects.. A derivative of vitamin A can reverse or suppress an epithelial preneoplasia, lending further support to the notion that chemoprevention of human cancer is feasible.

Topics: Administration, Intravaginal; Adult; Antineoplastic Agents; Biopsy; Female; Humans; Logistic Models; Remission Induction; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Patients with cervical precancerous lesions were double blindly divided into 2 groups and treated with either Retinamide II (RII) or Riboflavin (VB2) suppository. The suppository containing RII 20mg or VB2 5mg was given intravaginally daily for 100 days, divided into 2 courses. Clinical examination, papanicolaou cytology and tissue biopsy under colposcope were carried out before and after treatment. The results showed that the disappearance rates of precancerous lesions (DRPL) after the second course of treatment with RII or VB2 were 68.5% and 52.00%, and the overall response rates were 74.29% and 56.0%, respectively. 1 or 2 yrs after treatment, DRPL of the RII, Riboflavin and laser groups did not show significant difference (P > 0.05). The statistically significant differences were among the RII, Riboflavin, laser groups and antiphlogistic treatment group (P < 0.01). The results indicate that local application of RII and Riboflavin may serve as chemopreventive agents for cervical cancer.

Topics: Administration, Intravaginal; Double-Blind Method; Female; Follow-Up Studies; Humans; Precancerous Conditions; Riboflavin; Suppositories; Tretinoin; Uterine Cervical Neoplasms

Development of contingency recruitment plans in cancer chemoprevention research is as important as formulation of the initial plan. We found that requesting recruitment information from our initial CTCD subjects provided a framework for our contingency plan. The revised recruitment plan consisted of: 1) calling and sending letters to community gynecologists in private practice or affiliated with HMO's to explain the study and ask for referrals; 2) continued personal contact by the principal investigator with referring physicians; 3) sending thank you and follow-up letters to every physician who referred patients to the study; 4) soliciting Papanicolaou smear reports from HMO's if physicians of women with abnormal Papanicolaou smears gave permission to pathologists to release this information; 5) utilizing free media such as feature articles on the CTCD in local papers, public service announcements, and television "spots;" 6) continued use of brochures and posters printed for the initial recruitment effort; and 7) continued presentations to local professional physician and nurse groups about the study. Our contingency plan to date has provided us with 100% of our projected accrual. Thus, our recruitment methods have proved to be effective in accruing subjects for this cancer chemoprevention trial.

Topics: Double-Blind Method; Female; Humans; Randomized Controlled Trials as Topic; Surveys and Questionnaires; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Three cancer prevention trials are currently in their early phases at The Fred Hutchinson Cancer Research Center, the University of Washington School of Public Health and Community Medicine, and the Swedish Hospital. All 3 studies are randomized and placebo controlled. One large-scale study involves the daily administration of retinoids to persons with asbestos-related lung disease in an attempt toward reduction of their high risk for bronchogenic carcinomas and mesotheliomas. A second study involves administration of the same agents to long-term heavy smokers; a substantial feasibility and toxicity pilot study will precede a full-scale prevention trial. In the third trial, folic acid administration is evaluated in relation to the progression and regression of cervical dysplasia among women with abnormal Pap smears. We report here the rationale and the design for these 3 studies.

Topics: Asbestosis; Clinical Trials as Topic; Female; Humans; Lung Neoplasms; Male; Neoplasms; Random Allocation; Research Design; Retinoids; Smoking; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms; Washington

The prevention and/or suppression of preneoplastic foci in animals has been convincingly demonstrated with the use of a variety of compounds, including retinoids. We report here a phase I/II trial of beta-trans-retinoic acid delivered via a collagen sponge/diaphragm insert for neoplasia of the cervix. Eighteen patients with biopsy-proved grades 2 and 3 cervical intraepithelial neoplasia underwent four consecutive daily applications of trans-retinoic aicd to the cervix. Conization of the cervix was performed 4 weeks later. Significant vaginal toxicity in 10 of 18 (55%) patients was unrelated to the concentration or carrying media of the drug. A reduction in size of the intraepithelial lesion was noted by colposcopy in six of 18 (33%) patients with complete resolution of disease at conization in two patients. A new delivery system designed to avoid vaginal toxicity while defining the optimal concentration and dosing schedule for these patients is currently under investigation.

Topics: Administration, Topical; Cervix Uteri; Clinical Trials as Topic; Collagen; Colposcopy; Contraceptive Devices; Drug Evaluation; Female; Humans; Tretinoin; Uterine Cervical Neoplasms; Vagina

This study on all-trans retinoic acid was designed to explore its effect on the ability of Fra-1 to cervical cancer cell development. The results show that all-trans retinoic acid enhances the effect of Fra-1 on inhibiting cervical cancer proliferation and the glucose consumption, its effect on the loss of mitochondrial membrane potential, on the decreasing of lactic acid as well as ATP, and also influences the expression of MDM2/P53/P21 and LDHA.. The results show that the expression of Fra-1 is higher in all-trans retinoic acid-treated cervical cancer. Flow cytometry and kit detection show that all-trans retinoic acid can enhance the ability of Fra-1 to lose the mitochondrial membrane potential, inhibit the glucose consumption and the production of lactic acid as well as ATP. CCK8 and colony formation assays indicate that all-trans retinoic acid enhances the ability of Fra-1 to inhibit cell proliferation. In addition, through Western blot analysis, it was determined that P53 and P21 were up-regulated, and MDM2 and LDHA were down-regulated.. The overall results of the study strongly suggest that all-trans retinoic acid enhances the effect of Fra-1 on inhibiting cervical cancer proliferation and metabolism in vitro, and also influences the expression of MDM2/P53/P21 and LDHA.

Topics: Antineoplastic Agents; Cell Proliferation; Female; HeLa Cells; Humans; Proto-Oncogene Proteins c-fos; Signal Transduction; Tretinoin; Uterine Cervical Neoplasms

Tazarotene-induced gene 1 (TIG1) is a retinoic acid-inducible protein that is considered a putative tumor suppressor. The expression of TIG1 is decreased in malignant prostate carcinoma or poorly differentiated colorectal adenocarcinoma, but TIG1 is present in benign or well-differentiated tumors. Ectopic TIG1 expression led to suppression of growth in cancer cells. However, the function of TIG1 in cell differentiation is still unknown. Using a yeast two-hybrid system, we found that transmembrane protein 192 (TMEM192) interacted with TIG1. We also found that both TIG1A and TIG1B isoforms interacted and co-localized with TMEM192 in HtTA cervical cancer cells. The expression of TIG1 induced the expression of autophagy-related proteins, including Beclin-1 and LC-3B. The silencing of TMEM192 reduced the TIG1-mediated upregulation of autophagic activity. Furthermore, silencing of either TIG1 or TMEM192 led to alleviation of the upregulation of autophagy induced by all-trans retinoic acid. Our results demonstrate that the expression of TIG1 leads to cell autophagy through TMEM192. Our study also suggests that TIG1 and TMEM192 play an important role in the all-trans retinoic acid-mediated upregulation of autophagic activity.

Topics: Autophagy; Beclin-1; Cell Line, Tumor; Female; HeLa Cells; Humans; Membrane Proteins; Microscopy, Confocal; Microtubule-Associated Proteins; Transfection; Tretinoin; Uterine Cervical Neoplasms

Retinoic acid (RA) is a critical regulator of cell differentiation, proliferation, and apoptosis in various cell types. Recently, the RA-metabolizing enzyme CYP26A1 (cytochrome P450, family 26, subfamily A, polypeptide 1) has been shown to have an oncogenic function in breast carcinogenesis. However, the relevance of elevated CYP26A1 expression in human cancers remains to be clarified.. We immunohistochemically examined the expression of CYP26A1 in cervical squamous cell carcinoma (SCC) and its precursors, including low- and high-grade squamous intraepithelial lesions (LSIL and HSIL, respectively), as well as head and neck cancer (HNC). The association between CYP26A1 expression and a number of clinicopathological parameters was also evaluated.. CYP26A1 was not expressed in normal cervical epithelium. CYP26A1 expression was present in LSIL but limited to basal and parabasal cells. HSIL cases exhibited strong nuclear expression of CYP26A1 and mixed cytoplasmic expression patterns with widely distributed expression toward the epithelial surface. Importantly, strong cytoplasmic staining of CYP26A1 was observed in 19 of 50 (38%) patients with cervical SCC. Elevated expression of CYP26A1 was significantly associated with younger age (<50 years) and lymph node involvement (pN). Similarly, CYP26A1 was not expressed in non-neoplastic tissues of the head and neck, but strong cytoplasmic staining of CYP26A1 was observed in 52 of 128 (41%) HNC cases. Such strong CYP26A1 expression was significantly associated with the primary tumor stage of carcinomas (pT) and the pathological tumor-node-metastasis (pTNM) stage in HNC.. Our results indicated an elevated CYP26A1 expression in malignant and precancerous dysplastic lesions of the human cervix, which also increased with the progression of cervical squamous neoplasia. In addition, this report is the first to demonstrate the increased expression of CYP26A1 in HNC and its significant correlation with primary tumor growth. These data suggested that CYP26A1 overexpression might contribute to the development and progression of cervical malignancies and squamous neoplasia of the head and neck.

Topics: Carcinoma, Squamous Cell; Cytochrome P-450 Enzyme System; Disease Progression; Female; Head and Neck Neoplasms; Humans; Retinoic Acid 4-Hydroxylase; Tretinoin; Uterine Cervical Neoplasms

Aberrant histone acetylation plays an essential role in the neoplastic process via the epigenetic silencing of tumour suppressor genes (TSGs); therefore, the inhibition of histone deacetylases (HDAC) has become a promising target in cancer therapeutics. To investigate the correlation of histone acetylation with clinicopathological features and TSG expression, we examined the expression of acetylated H3 (AcH3), RARβ2, E-cadherin, and β-catenin by immunohistochemistry in 65 cervical squamous cell carcinoma patients. The results revealed that the absence of AcH3 was directly associated with poor histological differentiation and nodal metastasis as well as reduced/negative expression of RARβ2, E-cadherin, and β-catenin in clinical tumour samples. We further demonstrated that the clinically available HDAC inhibitors valproic acid (VPA) and suberoylanilide hydroxamic acid (SAHA), in combination with all-trans retinoic acid (ATRA), can overcome the epigenetic barriers to transcription of RARβ2 in human cervical cancer cells. Chromatin immunoprecipitation analysis showed that the combination treatment increased the enrichment of acetylated histone in the RARβ2-RARE promoter region. In view of these findings, we evaluated the antitumor effects induced by combined VPA and ATRA treatment in a xenograft model implanted with poorly differentiated human squamous cell carcinoma. Notably, VPA restored RARβ2 expression via epigenetic modulation. Additive antitumour effects were produced in tumour xenografts by combining VPA with ATRA treatment. Mechanistically, the combination treatment reactivated the expression of TSGs RARβ2, E-cadherin, P21 (CIP1) , and P53 and reduced the level of p-Stat3. Sequentially, upregulation of involucrin and loricrin, which indicate terminal differentiation, strongly contributed to tumour growth inhibition along with partial apoptosis. In conclusion, targeted therapy with HDAC inhibitors and RARβ2 agonists may represent a novel therapeutic approach for patients with cervical squamous cell carcinoma.

Topics: Animals; Antineoplastic Agents; beta Catenin; Cadherins; Carcinoma, Squamous Cell; Cell Line, Tumor; Disease Models, Animal; Epigenesis, Genetic; Female; Gene Expression Regulation, Neoplastic; Histone Deacetylase Inhibitors; Histone Deacetylases; Histones; Humans; Promoter Regions, Genetic; Protein Binding; Retinoid X Receptor beta; Transcriptional Activation; Tretinoin; Tumor Burden; Tumor Suppressor Proteins; Uterine Cervical Neoplasms; Valproic Acid; Xenograft Model Antitumor Assays

Epigenetic silencing of the tumor suppressor gene, RARβ2, through histone deacetylation has been established as an important process of cervical carcinogenesis. This pivotal role has led to the suggestion that a combination of retinoids selective for RARβ2 with histone deacetylase (HDAC) inhibitors may have therapeutic potential. Valproic acid (VPA), a HDAC inhibitor, has a critical role in the regulation of gene expression through histone acetylation and causes transformed cells to undergo growth arrest, differentiation, and apoptosis. Therefore, we hypothesized that the combination of VPA and ATRA could restore RARβ2 expression, thus resulting in enhanced anti-neoplastic activity in cervical cancer. Here, we show that VPA combined with ATRA led to hyperacetylation of histone H3 and a significant alteration of gene expression in cervical cancer cells, including RARβ2 gene expression, which was upregulated 50- to 90-fold. The combination therapy effectively inhibited the growth of cervical cancer cells more than the single agent treatment both in vitro and in vivo. The additive effects were associated with a significant upregulation of p21(CIP1) and p53 as well as a pronounced decrease in p-Stat3. Furthermore, the combined treatment led to cell cycle arrest predominantly at the G1 phase, and it preferentially induced cell differentiation rather than apoptosis in cervical cancer cells. The differentiation program was determined by the presence of E-cadherinmediated adhesion and activation of the PI3K/Akt pathway. Taken together, these results provide new insight into the mechanisms of enhanced antitumor activity of the HDAC inhibitor and ATRA regimen, thus offering a new therapeutic strategy for cervical cancer patients.

Topics: Antineoplastic Combined Chemotherapy Protocols; Cell Cycle; Cell Differentiation; Cell Line, Tumor; Female; HeLa Cells; Humans; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Receptors, Retinoic Acid; Signal Transduction; Tretinoin; Uterine Cervical Neoplasms; Valproic Acid

Histone deacetylase inhibitors (HDACIs) represent a new class of targeted anti-cancer agents and different other diseases, like muscular disorders. A number of studies have shown that extracellular signal-activated kinases can target chromatin-modifying complexes directly and regulate their function. The molecular connection between the dystrophin-associated protein complex (DAPC) and chromatin has been described, by showing that NO signaling regulates histone deacetylase (HDAC) activity and influences gene expression in different cell types. In present study, we investigated HDACs changes in HeLa cells undergoing growth inhibition and apoptosis, caused by HDACI BML-210 and retinoic acid (ATRA). Cell cycle analysis indicated that HeLa cell treatment with 20 and 30 μM concentration of BML-210 increased the proportion of cells in G0/G1 phase, and caused accumulation in subG1, indicating that the cells are undergoing apoptosis. We determined down-regulation of HDAC 1-5 and 7 after treatment with BML-210. Also, we demonstrated expression of different isoforms of alpha-dystrobrevin (α-DB) and other components of DAPC such as syntrophin, dystrophin, beta-dystrobrevin (β-DB) and NOS in HeLa cells after treatments. We determined changes in protein expression level of dystrophin, NOS1, α- and β-DB and in subcellular localization of α-DB after treatments with BML-210 and ATRA. In conclusion, these results suggest that HDACI BML-210 can inhibit cell growth and induce apoptosis in cervical cancer cells, what correlates with down-regulation of HDAC class I and II and changes in the DAPC expression levels. This can be important for identifying target proteins in DAPC signaling to HDACs, as a target of pharmacological intervention for treatment of muscular dystrophies and other diseases.

Topics: Anilides; Antineoplastic Agents; Apoptosis; Calcium-Binding Proteins; Cell Cycle; Cell Proliferation; Cell Survival; Down-Regulation; Drug Synergism; Dystrophin; Dystrophin-Associated Protein Complex; Dystrophin-Associated Proteins; Female; Gene Expression; Gene Expression Regulation, Neoplastic; HeLa Cells; Histone Deacetylase Inhibitors; Histone Deacetylases; Humans; Membrane Proteins; Muscle Proteins; Muscular Dystrophies; Nitric Oxide Synthase; Protein Transport; Tretinoin; Uterine Cervical Neoplasms

Retinoid X receptor-alpha (RXR alpha) fragments are known to be produced in some cancer cells by proteolytic cleavage. Previous finding that ligand binding domain (LBD) fragment of RXR alpha specifically inhibits retinoic acid receptor-gamma (RAR gamma) activity led us to investigate the functional role of RXR alpha LBD fragment in radiosensitization by retinoic acid (RA). Ectopic expression of RXR alpha LBD fragment in cells that do not have a detectable endogenous RXR alpha LBD fragment, blocked synergistic radiosensitizing action of RA, as determined by growth inhibition, cell death and colony formation assays. However, H460 cell, which has an endogenous RXR alpha LBD fragment, was not radiosensitized by RA regardless of the ectopic RXR alpha LBD fragment expression. These results were paralleled with the pattern of p21 Waf1/Cip1 induction by the treatment of RA in combination with ionizing radiation (IR). Taken together, we hypothesize that the RXR alpha LBD fragment may act as a negative regulator of radiosensitizing effect of RA by restricting the RAR gamma-mediated biological response to RA.

Topics: Apoptosis; Blotting, Western; Cell Proliferation; Colony-Forming Units Assay; Combined Modality Therapy; Cyclin-Dependent Kinase Inhibitor p21; Drug Synergism; Female; Humans; Ligands; Radiation-Sensitizing Agents; Radiation, Ionizing; Retinoid X Receptor alpha; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Although oncogenic human papillomavirus (HPV) infections have been established as the necessary cause of cervical cancer, most HPV infections are transient and rarely progress to squamous cervical lesions. The activity of HPV is tightly associated with epithelial cell differentiation; therefore, regulators of differentiation, such as retinoic acid (RA), have been considered targets for the prevention of HPV-associated squamous intraepithelial lesion (SIL) development. The purpose of this study was to determine the association between circulating RA and early events in cervical carcinogenesis, specifically type-specific HPV clearance and SIL detection. Archived blood samples from 643 women participating in the Ludwig-McGill Cohort in São Paulo, Brazil, were analyzed by high-pressure liquid chromatography for three RA isomers (all-trans, 13-cis, and 9-cis-RA). A type-specific HPV clearance event was defined as two consecutive visits negative for an HPV type during follow-up for 364 HPV-positive women. Among the 643 women in this analysis, 78 were diagnosed with incident SIL. The probability of clearing an oncogenic HPV infection was not significantly different across RA isomer quartiles. There was a suggestion that increasing all-trans-RA increased the rate of nononcogenic HPV clearance (P-trend = 0.05). There was no association observed between serum RA levels and incident SIL. Our results suggest that elevated circulating RA isomer levels do not increase the rate of HPV clearance or reduce the risk of incident SIL. The role of RA in the inhibition of HPV-induced carcinogenesis, as shown in vitro, lacks confirmatory evidence within epidemiologic studies among women.

Topics: Adolescent; Adult; Alphapapillomavirus; Biological Transport; Brazil; Carcinoma, Squamous Cell; Cohort Studies; Female; Humans; Incidence; Middle Aged; Papillomavirus Infections; Risk Factors; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms; Viral Load; Young Adult

Cdk5 is a small serine/threonine protein kinase which belongs to Cdk family. Unlike other Cdk members, so far Cdk5 is known to be irrelevant in cell cycle. Cdk5 kinase activity is regulated by binding with its activator, p35. Our previous results indicate that CdkS and p35 are involved in drugs-induced apoptosis of prostate cancer cells. Retinoic acid (RA) is one of the vitamin A-related compounds. Because of its potency on biological functions, it has been widely studied in its novel actions including the ability to inhibit cancer cell growth and to induce apoptosis. Here, we report that RA treatment decreased the growth of human cervical cancer cell line, HeLa, and Cdk5 contributed to this effect. The involvement of Cdk5 in RA-reduced cell survival was performed by treatments of Cdk5 inhibitor and siRNA. We further identified that RA-induced growth inhibition was partly correlated to Cdk5 activity-related apoptosis by detecting cell cycle distribution of sub G1 phase and the signals of Annexin V staining. In addition, our results also indicated that Cdk5 activity was involved in RA-induced HeLa apoptosis by detecting cleavages of caspase-3 and its substrate, PARP (poly (ADP-ribose) polymerases) Interestingly, the nuclear localizations of Cdk5 and p35 proteins were increased by RA treatment, which, again, suggests the involvement of Cdk5 and p35 in RA-induced apoptotic effects. In conclusion, we provide evidence to suggest that Cdk5 and p35 might play important roles in RA-induced HeLa apoptosis.

Topics: Adaptor Proteins, Signal Transducing; Antineoplastic Agents; Apoptosis; Caspase 3; Cell Cycle Proteins; Cell Survival; Cyclin-Dependent Kinase 5; Female; G1 Phase; Growth Inhibitors; HeLa Cells; Humans; Purines; RNA, Small Interfering; Roscovitine; Tretinoin; Uterine Cervical Neoplasms

To investigate the combined effect of the major tea polyphenol, (-)-epigallocatechin gallate (EGCG) and retinoic acid (RA) on cervical adenocarcinoma.. Cell growth rate was examined after treatment for 4, 7 and 10 days with 0-100 microM EGCG and/or 1 microM RA in two cervical adenocarcinoma cell lines, HeLa and TMCC-1. The effect of EGCG treatment was examined for the induction of apoptosis by DNA ladder assay and caspase-3-related protease activity in cell lysate. Telomerase activity was detected by stretch PCR telomere extension assay. hTERT expression levels were quantified by a real-time PCR system.. Combining EGCG and RA increased the antiproliferative effect in adenocarcinoma cell lines, whereas EGCG or RA treatment alone caused a less sensitive response in these cells. Neither EGCG nor RA treatment alone affected apoptosis and telomerase activity. The combination treatment of EGCG and RA induced apoptosis and inhibited telomerase activity in adenocarcinoma cell lines. These results were consistent with those of an antiproliferative effect of EGCG and/or RA in cervical adenocarcinoma cells.. Our data suggest that EGCG and RA combined to prevent the carcinogenesis of cervical adenocarcinoma, induce apoptosis and inhibit telomerase activity. The treatments of combining EGCG and RA may be effective in preventing or treating cervical adenocarcinoma.

Topics: Adenocarcinoma; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Caspase 3; Catechin; Cell Growth Processes; Cell Line, Tumor; Dose-Response Relationship, Drug; Drug Synergism; Female; HeLa Cells; Humans; Telomerase; Tretinoin; Uterine Cervical Neoplasms

In uterine cervical cancer, certain oncogenic HPV types are considered as key etiologic factor. But the progression of HPV associated cervical precancerous lesions depends on many other factors such as oncogenes, immune system, anti-viral factors etc. This study is therefore focused on the effect of an important dietary anti-viral factor called All Trans Retinoic Acid (ATRA) on the development of HPV associated cervical cancer as it is found higher in poor socioeconomic people.. We analyzed a total population of 130 including control subjects who have no complaints of uterine cervical lesions and the HPV-6/11, 16/18 infected cases of low grade squamous intraepithelial lesions [SIL], high grade squamous intraepithelial lesions [HSIL], and invasive cancers, for serum ATRA level. This study also focused to find out the association of serum ATRA level with the proliferation status in terms of proliferating cell nuclear antigen (PCNA) expression as it is an anti-proliferation agent and with the grades of cervical lesions, using SPSS statistical package.. The results showed a highly significant negative association for serum ATRA level with different stages of cervical lesions (F = 3.305; P = 0.000) by one-way ANOVA and with intensity of PCNA expression (r = -0.825; P < 0.01) by Pearson's correlation test. A highly significant association was observed for the PCNA expression with the grades of cervical lesions too (F = 37.89; P = 0.000). Further, we found from our data that all the invasive cancer cases were infected with HPV-16/18 and none with HPV-6/11. Hence, we analyzed the association of serum ATRA level with HPV-16/18 infected preinvasive cases in developing invasiveness, by Fisher's Exact Test, using Graph Pad Prism as shown in Table 1. The results show an odds ratio (OR) of 36.93 and a relative risk (RR) of 4.99 with an 95% interval being 2.896 to 8.603, which is significant at the level of P = 0.0001 for the reduced [<0.6 mug/ml] serum ATRA level in developing invasive cancer in HPV-16/18 infected preinvasive cases.. All these results suggest that the serum ATRA level highly influences the progression of cervical lesions to invasive cancer and can be therefore aimed as a marker for progression in combination with HPV-16/18, which helps to enhance the modalities of therapy towards cost effectiveness.

Topics: Adult; Cell Growth Processes; Cell Transformation, Viral; Female; Humans; Middle Aged; Neoplasm Staging; Papillomaviridae; Papillomavirus Infections; Polymerase Chain Reaction; Proliferating Cell Nuclear Antigen; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Human papillomavirus (HPV) infection is believed to be the central cause of cervical cancer. The viral proteins E6 and E7 from high-risk HPV types prevent cells from differentiating apoptosis and inducing hyperproliferative lesions. Human cervical carcinoma HeLa cells contain integrated human papillomavirus type 18 (HPV-18). Retinoic acid (RA) is a key regulator of epithelial cell differentiation and a growth inhibitor in vitro of HeLa cervical carcinoma cells. Cellular responses to RA are mediated by nuclear retinoic acid receptors (RARs) and retinoid X receptors. On the other hand, histone deacetylase inhibitors have been shown to be chemopreventive agents for the treatment of cancer cells. In this article, we have examined the antiproliferative effect of RA and histone deacetylase inhibitor BML-210 on HeLa cells, and particularly the effects on protein expression that may be involved in the cell cycle control and apoptosis. Our data suggest that a combination of RA and BML-210 leads to cell growth inhibition with subsequent apoptosis in a treatment time-dependent manner. We confirm that BML-210 alone or in combination with RA causes a marked increase in the level of p21. The changes in the p53 level are under the influence of p38 phosphorylation. We also discovered that the histone deacetylase inhibitor BML-210 causes increased levels of anti-apoptotic protein Bcl-2 and phosphorylated p38 MAP Kinase; the latter link in cell cycle arrest with response to extracellular stimuli. Our results suggest that RA and BML-210 are involved in different signaling pathways that regulate cell cycle arrest and lead to apoptosis of HeLa cells.

Topics: Anilides; Cell Proliferation; Female; Growth Inhibitors; HeLa Cells; Histone Deacetylase Inhibitors; Histone Deacetylases; Humans; Tretinoin; Uterine Cervical Neoplasms

Current therapy for cervical cancer includes radiation therapy. Retinoic acid (RA) can increase the sensitivity of cervical cancer cell lines to radiation. The mechanism of this sensitization may not involve the p53 protein because the human papillomavirus (HPV) E6 protein, which is present in the majority of cervical cancers, promotes p53 degradation. The objective of this study was to determine if p53 is involved in the mechanism of RA radiosensitization.. The effects of radiation on cervical (SiHa, CC-1, and C33a) and vulvar (SW962) cancer cell lines under various experimental conditions were evaluated using clonogenic, Coulter Counter, electrophoretic mobility shift (EMSA) and a multi-probe RNase protection assay of p53-inducible genes.. RA (5 microM 9-cis-RA) radiosensitized the SiHa and CC-1 cell lines that contain HPV-degraded p53, but did not radiosensitize the SW962 cell line, which is HPV negative and contains wild-type p53, nor the C33a cell line, which contains mutant p53 (R273C). Expression of mutant p53 (R273H) in SiHa cells increased the growth rate, but did not prevent RA-induced differentiation or radiosensitization at clinically relevant doses. Inhibition of p53 transactivation with pifithirin alpha did not prevent RA radiosensitization of SiHa at 5 Gy. RA repressed c-fos mRNA expression in control and irradiated SiHa cultures, but did not repress bcl-x(L), p53, GADD45, p21, bax, bcl-2, or mcl-1 mRNA expression.. The mechanism of RA radiosensitization does not require functional p53 and may involve c-fos in cervical cancer cell lines.

Topics: Alitretinoin; Benzothiazoles; Cell Differentiation; Cell Line, Tumor; Cell Proliferation; Combined Modality Therapy; DNA Damage; Dose-Response Relationship, Radiation; Female; Genes, fos; Humans; Radiation-Sensitizing Agents; Thiazoles; Toluene; Transfection; Tretinoin; Tumor Suppressor Protein p53; Uterine Cervical Neoplasms

This review contrasts the planned and actual recruitment and retention efforts for a cervical cancer prevention study within a predominantly underserved population. Recruitment was a primary obstacle to trial progression and multiple strategies to improve recruitment were implemented to meet objectives. The actual recruitment strategies were expansion to five geographically distinct clinical sites, use of nurse practitioners focused primarily on patient issues, extremely flexible study hours and location, honorariums, support for transportation and child care, and creativity in maintaining contact with study participants. With these strategies, 90% of eligible patients consented to participate in the study.

Topics: Adult; Biopsy; Colposcopy; Community-Institutional Relations; Female; Hospitals, University; Hospitals, Urban; Humans; Midwestern United States; Needs Assessment; Nurse Practitioners; Nurse's Role; Patient Compliance; Patient Dropouts; Patient Selection; Poverty; Randomized Controlled Trials as Topic; Research Personnel; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms; Women

All-trans retinoic acid (ATRA) suppresses growth of cervical dysplasias in vivo, although the sensitivity to retinoids is frequently lost during cervical carcinogenesis. It has been suggested that prolonged treatment or use of higher doses of retinoids might offer favorable response rates. We found SiHa cervical squamous carcinoma cells that were virtually resistant to ATRA-induced growth-inhibitory effects at physiological doses (10(-7 to) 10(-6) M) to be more responsive at pharmacological doses (10(-5 to) 10(-4) M). The growth inhibition by high-dose (10(-4) M) ATRA was associated with a sustained activation of interferon regulatory factor 1 (IRF-1), while a low dose (10(-6) M) of ATRA activated IRF-1 only transiently. Antisense IRF-1 inhibited the high-dose (10(-4) M), ATRA-mediated growth arrest; forced expression of IRF-1 caused a significant reduction in cell growth. High-dose (10(-4) M) ATRA increased binding of NF-kappaB and STAT1 proteins to sequences that originated from the IRF-1 promoter region, while low-dose (10(-6) M) ATRA induced only NF-kappaB binding. A delayed tyrosine phosphorylation of the signal transducer and activator of transcription-1 (STAT1) was observed after high-dose (10(-4) M) but not low-dose (10(-6) M) ATRA treatment. In agreement with this, induction of IRF-1 mRNA by ATRA was only modest and transient in a STAT1 knockout cell line, suggesting the importance of STAT1 in sustained IRF-1 expression. Our data showed that ATRA is capable of inducing dose-dependent cellular changes, which might be appropriate to overcome resistance to retinoids that frequently develops during cervical carcinogenesis.

Topics: Antineoplastic Agents; Carcinoma, Squamous Cell; Cell Division; DNA-Binding Proteins; Dose-Response Relationship, Drug; Female; Gene Expression Regulation, Neoplastic; Humans; Interferon Regulatory Factor-1; NF-kappa B; Oligonucleotides; Phosphoproteins; Phosphorylation; Promoter Regions, Genetic; STAT1 Transcription Factor; Trans-Activators; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Telomerase, a ribonucleoprotein complex of hTERT, hTR, and TP1, has been reported to be associated with carcinogenesis and multidrug resistance (MDR). This study used our in vitro human cervical multistep carcinogenesis/MDR model system in which normal human ectocervical and endocervical (HEN) cells were immortalized by HPV18 or 16, respectively, and subsequently transformed. The first evidence was found that immortalization and telomerase activation were correlated with increased expression specifically of two of the hTERT alternatively spliced mRNAs, one encoding wild-type protein containing the full-length functional reverse transcriptase (RT) region and one encoding a defective RT protein. Expression of neither hTERT mRNA containing full-length functional or defective RT motif was affected by transformation/MDR. All-trans-retinoic acid (ATRA) treatment of HPV-immortalized HEN-16-2 cells and transformed/MDR HEN-16-2/CDDP cells inhibited telomerase activity and downregulated expression of hTERT mRNAs containing full-length functional and a defective RT motif, but there were no changes in hTR and TP1 expression. Moreover, ATRA inhibited cell growth rate of HEN-16-2 and HEN-16-2/CDDP cells equally. These results provided the first evidence that ATRA equally in both immortalized and transformed/MDR cell lines inhibits telomerase activity and downregulates expression, but not splicing, of hTERT, and this is correlated with cell growth rate inhibition; the potential is implicated for applying ATRA to hTERT-targeted treatment of cervical cell carcinogenesis/MDR.

Topics: Alternative Splicing; Antineoplastic Agents; Carrier Proteins; Cell Division; Cell Line, Transformed; Cell Transformation, Neoplastic; Cells, Cultured; Cervix Uteri; DNA-Binding Proteins; Down-Regulation; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Enzyme Activation; Female; Gene Expression Profiling; HeLa Cells; Humans; Models, Biological; RNA-Binding Proteins; Telomerase; Tretinoin; Uterine Cervical Neoplasms

The aim of this study was to determine whether receptor-dependent and receptor-independent retinoids sensitize cervical cancer cells to clinically relevant doses of concurrent radiation and cisplatin.. The clonogenic assay was performed on SiHa cervical carcinoma cultures treated with 5 microM 9-cis-retinoic acid (RA) or 3 microM 4-HPR for 3 days prior to and following concurrent treatment with 3 microM cisplatin and 2 Gy of Co(60) radiation.. Neither 9-cis-RA nor 4-HPR significantly decreased survival for radiation only or cisplatin only (t test: P < 0.05), but both significantly decreased survival of cultures receiving concurrent chemoradiation (t test: 9-cis-RA P = 0.045; 4-HPR P = 0.027).. Both receptor-dependent and receptor-independent retinoids enhance concurrent chemoradiation effects in vitro.

Topics: Alitretinoin; Antineoplastic Combined Chemotherapy Protocols; Cell Survival; Cisplatin; Combined Modality Therapy; Drug Synergism; Female; Fenretinide; Humans; Receptors, Retinoic Acid; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

To investigate and compare the efficacy of all-trans retinoic acid (RA) and/or interferon-alpha (IFN-alpha) on premalignant and malignant models of cervical cancer.. Cell growth rate was examined after treatment for 4, 7, and 10 days with RA and/or IFN-alpha of human papillomavirus type 18 (HPV 18)-immortalized endo- and ectocervical cells, nontransformed serum-adapted cells, transformed cells, three adenocarcinoma, and three squamous cell carcinoma cell lines. The effect on epithelial differentiation by RA and IFN-alpha was examined in organotypic culture. Induction of apoptosis was examined by modified terminal transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling (TUNEL) and DNA fragmentation.. Cell growth rate was inhibited by RA, 84-96% in HPV 18-immortalized endocervical cells, SiHa, and ME180, 0% in OMC-4, and 18-62% in other cell lines; and by IFN-alpha about 75% in SiHa and ME180 and 14-40% in the other cell lines. Combining RA and IFN-alpha increased the antiproliferative effect in premalignant cell lines and some cancer cell lines except OMC-4, SiHa, and HT-3. In rafts, RA treatment reversed human endocervical cell metaplasia and HPV 18-immortalized endo- and ectocervical cell dysplastic epithelial differentiation. Interferon-alpha, not RA, treatment of HPV 18-immortalized endo- and ectocervical cells induced apoptosis.. Cell growth inhibition by treatment with RA, IFN-alpha, and their combination differentially depends on treatment type and time, cell origin, cell line, and oncogenic state. In a premalignant model of cervical carcinoma, RA reduces dysplastic differentiation and IFN-alpha induces apoptosis. These data confirm that these treatments may be effective for preventing or treating premalignant cervical lesions.

Topics: Adenocarcinoma; Antineoplastic Agents; Apoptosis; Carcinoma, Squamous Cell; Cell Differentiation; Cell Division; Cell Line, Transformed; Cervix Uteri; Female; Humans; Interferon-alpha; Papillomaviridae; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Retinoids and interferons have been implicated in the growth regulation of cervical cancer cells. However, the molecular mechanisms are not fully defined. To analyze detailed mechanisms, HPV-positive (HeLa, CaSki), HPV-negative (C33A, HT-3) and non-cervical Cos-1 cell lines were treated with I microM all-trans-retinoic acid (RA) and/or 10 ng/ml interferon-gamma (IFN-gamma). The growth of RA-treated HeLa cells was less effectively suppressed than that of IFN-gamma-treated ones. A combination of RA and IFN-gamma leads to an additive antiproliferative effect on the cell growth. CaSki cell growth was also inhibited by IFN-gamma but was little stimulated by RA treatment, and the IFN effect was attenuated when IFN-gamma was combined with RA. HPV-negative C33A and HT-3 cells, which are defective in p53 and Rb, and Cos- 1 cells were weakly or not responsive to all combined treatments. The molecular mechanism underlying the differential effects of RA/IFN on HeLa and C33A cells was investigated. Combined RA/IFN-gamma treatment caused a marked increase in the level of IFN regulatory factor-1 (IRF-1) in HeLa, whereas no induction of IRF-1 was observed in C33A, consistent with the findings that IFN signaling is functional in HeLa but is defective in C33A cells. The increase of p53 in HeLa cells might account for the down-regulation of HPV-18 E6 gene expression by RA/IFN-gamma. Furthermore, RA/IFN-gamma treatment resulted in the concurrent induction of p21WAF1 CDK inhibitor and dephosphorylation of Rb protein. Transient co-expression of IRF-1 and p53 led to the cooperative activation of the p21WAF1 promoter. Our results indicate that 2 transcription factors, increased in response to RA/IFN-gamma, cooperate functionally to regulate the cell cycle through the activation of a common p21WAF1 gene in HeLa cells.

Topics: Antineoplastic Combined Chemotherapy Protocols; Blotting, Western; Cell Division; DNA-Binding Proteins; DNA, Neoplasm; Enzyme-Linked Immunosorbent Assay; Female; HeLa Cells; Humans; Interferon Regulatory Factor-1; Interferon-gamma; Phosphoproteins; Reverse Transcriptase Polymerase Chain Reaction; Time Factors; Transcription Factors; Tretinoin; Tumor Cells, Cultured; Tumor Suppressor Protein p53; Uterine Cervical Neoplasms

Type I Interferon (IFN) and all-trans retinoic acid (RA) inhibit cell proliferation of squamous carcinoma cell lines (SCC). Examinations of growth-affected cell populations show that SCC lines ME-180 and SiHa treated with IFN-beta undergo a specific slower progression through the S phase that seems to trigger cellular death. In combination treatment RA potentiates IFN-beta effect in SCC ME-180 but not in SiHa cell line, partially resistant to RA antiproliferative action. RA added as single agent affects cell proliferation differently by inducing a slight G1 accumulation. The IFN-beta-induced S phase lengthening parallels the increased expression of PML, a nuclear phosphoprotein specifically up-regulated at transcriptional level by IFN, whose overexpression induces cell growth inhibition and tumor suppression. We report that PML up-regulation may account for the alteration of cell cycle progression induced by IFN-beta in SCC by infecting cells with PML-PINCO recombinant retrovirus carrying the PML-3 cDNA under the control of the 5' LTR. In fact PML overexpression reproduces the IFN-beta-induced S phase lengthening. These findings provide important insight into the mechanism of tumor suppressing function of PML and could allow PML to be included in the pathways responsible for IFN-induced cell growth suppression.

Topics: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Squamous Cell; Cell Division; DNA, Neoplasm; Female; Gene Expression Regulation, Neoplastic; Growth Inhibitors; Humans; Interferon Type I; Neoplasm Proteins; Nuclear Proteins; Promyelocytic Leukemia Protein; Protein Isoforms; Recombinant Proteins; S Phase; Transcription Factors; Tretinoin; Tumor Cells, Cultured; Tumor Suppressor Proteins; Up-Regulation; Uterine Cervical Neoplasms

We have evaluated the tumor tissue pO2 in cervical cancers during radiotherapy with special emphasis on the course of the pO2 in primarily hypoxic tumors and in patients treated with radiotherapy plus 13-cis-retinoic acid/interferon-alpha-2a.. From June 1995 through April 1997, 49 patients with squamous cell carcinoma FIGO IIB-IVA of the cervix who were treated with definitive radiotherapy with curative intent underwent polarographic measurement of tumor tissue pO2 with an Eppendorf pO2-histograph prior to and during radiation treatment. Radiotherapy consisted of external irradiation with 50.4 Gy in 28 fractions of 1.8 Gy plus high dose rate (HDR) brachytherapy. Twenty-two patients had additional treatment with 13-cis-retinoic acid (cRA, isotretinoin) and interferon-alpha-2a (IFN-alpha-2a). Therapy with cRA/IFN in these patients started 2 weeks before radiotherapy; during this induction period, cRA was administered in a dosage of 1 mg per kilogram body weight orally daily and IFN-alpha-2a in a dosage of 6x10(6) I.U. subcutaneously daily. After start of external radiotherapy (XRT), cRA/IFN was continued concomitantly with radiotherapy in reduced doses (0.5 mg cRA per kg body weight orally daily plus 3x10(6) I.U. IFN-alpha-2a subcutaneously three times weekly until the end of the radiation treatment). PO2 measurements were performed prior to radiotherapy, at 20 Gy, and at the end of radiotherapy.. A poor oxygenation defined as a median pO2 of 10 mm Hg or less was present in 15/38 tumors (39%) in which measurements prior to any treatment were done. Low pO2 readings below 5 mm Hg were present in 70% of all tumors prior to treatment. In 13 of 15 hypoxic tumors, pO2 measurements at 19.8 Gy were performed. In these tumors, a significant increase of the median pO2 from 6.0+/-3.1 mm Hg to 20.7+/-21.2 mm Hg was found, p<0.01. The increase in the median pO2 was more pronounced in patients with radiotherapy plus additional cRA/IFN treatment as compared to patients treated with irradiation alone (median pO2 raised from 7.0+/-3.5 mm Hg to 40.9+/-21.3 mm Hg versus 5.7+/-3.1 mm Hg to 14.7+/-17.9 mm Hg). In a multivariate analysis, both the effect of radiation dose (pretreatment versus 19.8 Gy) and the type of treatment (XRT alone versus XRT plus cRA/IFN) had significant impact on the pO2 (P = 0.003 and p = 0.04). In patients with well-oxygenated tumors (pretreatment median pO2>10 mm Hg), 20/23 (87%) achieved a clinically complete response. In patients with primarily hypoxic tumors, 6/6 patients whose primarily hypoxic tumors showed an increase of the median pO2 above 10 mm Hg at 19.8 Gy achieved a complete remission (CR). In contrast, only 4/7 patients with a low pretreatment and persisting low median pO2 achieved a CR.. There are evident changes in the oxygenation of cervical cancers during a course of fractionated radiotherapy. In primarily hypoxic tumors, a significant increase of the median pO2 was found. An additional treatment with cis-retinoic acid/interferon further improved the oxygenation. An impact of the different patterns of oxygenation on local control is to be evaluated.

Topics: Adult; Aged; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Squamous Cell; Cell Hypoxia; Cisplatin; Combined Modality Therapy; Dose Fractionation, Radiation; Female; Humans; Interferon alpha-2; Interferon-alpha; Middle Aged; Oxygen; Partial Pressure; Recombinant Proteins; Tretinoin; Uterine Cervical Neoplasms

Retinoic acid (RA) has been shown to radiosensitize some tumor cell lines. RA regulates gene expression through nuclear receptors that bind to retinoic acid response elements in gene promoters and that inhibit activator protein-1 (AP-1) transcription factor activity.. The aim of this study was to determine if the mechanism of radiosensitization of the CC-1 human cervical carcinoma cell line by 9-cis-RA (9cRA) involves repression of AP-1 activity.. The CCA reporter cell line was established from CC-1 by permanent transfection with the ColCAT reporter plasmid which consists of the chloramphenicol acetyltransferase (CAT) gene under the control of the AP-1-responsive collagenase gene promoter. CCA cultures were treated with various combinations of 9cRA, 60Co radiation, and an AP-1 inducer called O-tetradecanoylphorbol 13-acetate (TPA). Cultures were then evaluated in parallel for CAT expression as a measure of AP-1 activity and for clonogenic survival as a measure of radiosensitization.. The CCA reporter line exhibited a radiation dose-responsive induction of AP-1 activity that was decreased by 5 microM 9cRA and increased by 50 ng/ml TPA. Simultaneous treatment with TPA and 9cRA prevented 9cRA repression of AP-1 and resulted in AP-1 activity above basal level. The 9cRA radiosensitized CC-1 cultures with a dose modification factor of 1.5. The survival of cultures treated simultaneously with TPA and 9cRA was statistically identical to that of cultures treated with 9cRA alone.. Although TPA prevented AP-1 repression by 9cRA, it did not prevent radiosensitization in CCA cultures, therefore the mechanism of radiosensitization of CCA by 9cRA is independent of AP-1 repression.

Topics: Alitretinoin; Antineoplastic Agents; Dose-Response Relationship, Drug; Female; Gene Expression Regulation, Neoplastic; Humans; Radiation-Sensitizing Agents; Tetradecanoylphorbol Acetate; Transcription Factor AP-1; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Cell surface receptors have been the subject of intensive investigations over the past few decades. One very important group of receptors on the cell surface is the "integrin" receptors which bind to extracellular matrix (ECM) proteins. Because of integrin's importance in cellular growth, development, and morphology the role of integrin receptors in cellular transformation, malignant growth, and metastasis has received wide attention. In this article we report on the effect of all-trans-retinoic acid (ATRA) on (a) the integrin family of cell surface receptors, (b) collagenase enzyme activity, and (c) invasive potential in human cervical cancer (SiHa) cells. A comparative cell adhesion assay clearly showed that ATRA affects the cell surface integrin receptors against different ECM proteins in a dose- and time-dependent manner. The binding of SiHa cells to ECM proteins (fibronectin, vitronectin, laminin, collagen IV) was drastically reduced when cells were treated with ATRA at 10 microM for 96 h in culture. Interestingly, when ATRA-treated (10 microM, 96 h) SiHa cells were allowed to grow for 15 days in ATRA-free complete medium the binding of SiHa cells to fibronectin and vitronectin was inhibited, even after 15 days of drug withdrawal, whereas cell adhesion to laminin and collagen IV returned to normal within 3-7 days. The comparative immunoprecipitation of two cell surface integrin receptors (alpha5beta1 and alphavbeta3) shows the effect of ATRA on the expression of alpha5, alphav, and beta1 subunits. In ATRA-treated SiHa cells the cell surface expression of the alphav subunit (in alphavbeta3 receptor) is much less than in untreated SiHa cells. In the case of the alpha5beta1 integrin receptor ATRA treatment caused a significant reduction in the expression of both alpha5 and beta1 subunits on the cell surface. Comparative zymography clearly demonstrated the inhibitory effect of ATRA on collagenase enzyme activity. Interestingly, the effect was irreversible, even after 15 days of culture in ATRA-free medium. The assay of the invasive potential of ATRA-treated and untreated SiHa cells in Boyden's invasion chamber demonstrated that ATRA treatment (10 microM, 96 h) inhibits the invasive potential of SiHa cells. The effect was not reversible even after 15 days of culture in ATRA-free medium. In conclusion, our observations indicate that ATRA has an inhibitory effect on the expression of SiHa cell surface integrin receptors and collagenase enzyme activity. The

Topics: Cell Adhesion; Cells, Cultured; Collagenases; Female; Humans; Integrins; Neoplasm Invasiveness; Tretinoin; Uterine Cervical Neoplasms

Retinoids and interferons are important regulators of human epithelial cell differentiation and have been successfully used in the clinical treatment of HPV-involved cervical cancer. In this study, 2 HPV-positive human cervical-carcinoma cell lines were analyzed for their surface expression of MHC-Class-I, MHC-Class-II and ICAM-1 surface antigens before and after exposure to all-trans retinoic acid, interferon-gamma and the combination of the 2 compounds. In addition, the effects on HLA-Class-I-mRNA expression were evaluated after such treatments. Both cell lines expressed MHC-Class-I molecules, and their levels were markedly up-regulated after exposure to IFN-gamma. Similarly, MHC-Class-II and ICAM-1 antigens were either induced or significantly up-regulated by IFN-gamma. Exposure to all-trans retinoic acid was also able to significantly increase the expression of MHC-Class-I and ICAM-I antigens as compared with untreated tumor cells. However, unlike IFN-gamma, retinoids were not able to induce the expression of HLA-Class-II surface antigens. Exposure to the combination of IFN-gamma plus retinoic acid significantly up-regulated (in an additive manner) HLA-Class-I and ICAM-1 molecules as compared with the levels obtainable after exposure to IFN-gamma alone. Finally, Northern-blot analysis of HLA-Class-I-mRNA expression confirmed that the activity of both of these biologic response modifiers was at transcriptional level. These data indicate that the combination of these modalities could induce an additive effect on the expression of immunologically important surface antigens on human cervical-cancer cells. These findings, together with the known anti-proliferative effects mediated by retinoids and IFN-gamma on tumor cells, further support the combination of these agents in the treatment of pre-invasive and invasive human cervical cancer.

Topics: Blotting, Northern; Female; Histocompatibility Antigens Class I; Humans; Intercellular Adhesion Molecule-1; Interferon-gamma; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Several lines of evidence have demonstrated that IFNs could be relevant in the treatment of certain neoplastic diseases such as carcinomas. In particular, IFN-alpha, in addition to the anti-proliferative and cytostatic effects, was demonstrated to be capable of inducing cell death by apoptosis both in vivo and in vitro. Numerous protocols have also been proposed which consider the association of IFN-alpha with other drugs. Among these are retinoids, a class of compounds capable of inducing inhibition of cell growth and differentiation. We address the question here by analyzing the role of cell adhesion in susceptibility to IFN-alpha, RA and their combination of a human cell line derived from a squamous carcinoma of the cervix, the Bcl-2-negative SiHa cell line. In this context, cytoskeleton components and several surface molecules playing a role in cell substrate and cell-to-cell relationships have been evaluated. We found that RA treatment is capable of improving stress fiber formation, decreasing cell detachment and increasing cell-adhesion capability. However, no variations in the ability to adhere to specific extracellular-matrix molecules were found in RA-treated cells. No quantitative changes were detected in integrins involved as receptors for extracellular matrix molecules (VLAI-VLA5) or in other cell-adhesion-associated molecules (e.g., CD44). By contrast, 2 important molecules involved in cell-adhesion processes appeared to be up-regulated by RA exposure: focal adhesion kinase and E-cadherin, involved in adhesion plaque formation and cell-to-cell contacts, respectively. Keeping in mind the importance of adhesion properties in the cell-growth pathway, our findings could be of interest in the study of carcinoma-cell proliferation and metastatic potential.

Topics: Cadherins; Carcinoma, Squamous Cell; Cell Adhesion; Cell Adhesion Molecules; Cell Division; Female; Focal Adhesion Kinase 1; Focal Adhesion Protein-Tyrosine Kinases; Humans; Interferon-alpha; Protein-Tyrosine Kinases; Proto-Oncogene Proteins c-bcl-2; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Retinoids are a class of compounds structurally related to vitamin A which have been found to be active agents experimentally as well as clinically in the prevention and treatment of cervical cancer. Recent data have suggested that in addition to their key regulatory role during epithelial cell differentiation, they could also contribute to enhanced cellular and humoral immunity against tumor cells. Hsp gp96 molecules have recently been implicated in the presentation of tumor and viral antigens. A number of key elements in this pathway, including major histocompatibility complex (MHC) class I molecules as well as adhesion/co-stimulation molecules such as ICAM-1 have reported to be sensitive to retinoic acid up-regulation. In this study we analyzed at the transcriptional (Northern blot) and post-transcriptional levels (Western blot) the effects of retinoic acid on the expression of the tumor rejection antigen (heat shock protein gp96) in three human cervical carcinoma cell lines. Exposure of therapeutic doses of retinoic acid (i.e. 1 microM) significantly and consistently increased the expression of heat shock protein gp96 (Western blot analysis) on CaSki, SiHa and HT-3 cervical cancer cell lines. Northern blot analysis demonstrated that the increase in the amount of protein was due to the transcriptional upregulation of this gene. Taken together, our results show that retinoic acid can significantly increase the expression of yet another immunologically important cell molecule, the tumor rejection antigen heat shock protein gp96 in human cervical cancer. Such findings provide new information on the effects of retinoic acid on tumor cells and further support the role of retinoic acid as a powerful biologic response modifier.

Topics: Antigens, Neoplasm; Antigens, Surface; Antineoplastic Agents; Blotting, Northern; Blotting, Western; Female; Gene Expression Regulation, Neoplastic; Heat-Shock Proteins; Humans; Immunologic Factors; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Retinoids, including natural vitamin A and its analogs, have been closely studied as chemopreventive drugs. The mechanism of action of retinoids, however, is not completely understood. Our study evaluated the effects of all-trans (high affinity ligand for both RAR and RXR receptors) and 9-cis retinoic acid (binds only with RXR receptors) on E6-E7 transcription, cell proliferation, cell cycle distribution, and p53 expression in CaSki cells, a cell line derived from cervical carcinoma containing 600 copies of the HPV-16 genome. Using quantitative RT-PCR analysis, we found that CaSki cells treated with all trans retinoic acid (ATRA) for seven days had a remarkably low level of E6-E7 transcription at 10(-5) M to 10(-9) M concentrations. A smaller inhibitory effect was observed on the E6-E7 transcription at a concentration of 10(-5) M with only 9-cis retinoic acid. Flow cytometric analysis revealed that cells treated with both all trans and 9-cis RA showed an increase in the mean percentage (93.5% and 86.1% respectively) of cells in the G1 phase as compared to untreated CaSki cells (55%) and normal keratinocytes (58%). The percentage of cells in the S phase decreased from a mean percentage of 28 and 26.5 to 5.8 and 5, respectively, after treatment with all trans retinoic acid and 9-cis retinoic acid. An increase in the level of immunophenotypic expression of wild type p53 was also noted after treatment with all trans retinoic acid and 9-cis retinoic acid. All trans and 9-cis retinoic acid may act on highly proliferating tumor cells by initially arresting DNA synthesis and inducing G1 arrest. In addition, they may be inducing a p53 dependent cell cycle arrest and thus suggests that all-trans and 9-cis retinoic acid may have a cytostatic effect rather than a cytotoxic effect on CaSki cells. The increased expression of p53 positive cells and the inhibition of E6/E7 transcription after treatment with these retinoids may indicate the potential role of all trans and 9-cis retinoic acid as a cell cycle regulator and an antiviral chemoprevention agent.

Topics: Alitretinoin; Cell Cycle; Female; Flow Cytometry; Humans; Oncogene Proteins, Viral; Papillomavirus E7 Proteins; Polymerase Chain Reaction; Repressor Proteins; RNA, Messenger; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Radiation treatment is one of the most standardized and effective modalities for contemporary cervical cancer therapy. In addition, the radiation-potentiating effects of retinoic acid have been recently described. In order to investigate whether enhanced immunogenicity might be responsible for such potentiation, we have evaluated the effects of retinoic acid combined with high doses of gamma-irradiation on the expression of major histocompatibility complex (MHC) Class I and II and intercellular adhesion molecule-1 (ICAM-1) in human cervical carcinoma cell lines.. The expression of surface antigens (MHC Class I and II and ICAM-1) was evaluated by FACS analysis in untreated control cells and in cells following their exposure to retinoic acid, high doses of gamma-irradiation (i.e., 5000 and 10,000 cGy), or the combination of the two procedures.. HT-3 and SiHa cervical cancer cells expressed variable levels of MHC Class I and ICAM-1 antigens while Class II surface antigens were not detectable. Exposure to either 5000 or 10,000 cGy completely inhibited cell replication in both cell lines and significantly and consistently increased the expression of all surface antigens present on the cells prior to irradiation. Irradiation was unable to induce neoexpression of antigens previously not expressed by these cells (i.e., MHC Class II). In a similar fashion, retinoic acid was also able to significantly increase the expression of MHC Class I and ICAM-1 antigens when compared to untreated tumor cells but was not able to induce the expression of HLA Class II surface antigens. Exposure to the combination of radiation plus retinoic acid significantly upregulated HLA Class I and ICAM-1 molecules in an additive manner when compared to the levels obtainable with the exposure to radiation or retinoic acid alone.. These data indicate that the combination of these two treatments could induce an additive effect on the expression of immunologically important surface antigens in human cervical cancer cells. These findings, together with the powerful antiproliferative effect of retinoids and irradiation on tumor cells, suggest that the combined regimen may be a promising and more effective combination for the treatment of cervical cancer.

Topics: Antineoplastic Agents; Female; Histocompatibility Antigens Class I; Histocompatibility Antigens Class II; Humans; Intercellular Adhesion Molecule-1; Radiation Dosage; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Retinoids are a class of compounds that are structurally related to vitamin A and have been found to be effective in the prevention and treatment of cervical cancer. To investigate whether enhanced immunogenicity might be responsible for such efficacy, we evaluated the effects of retinoic acid on the expression of major histocompatibility complex class I and class II and intercellular adhesion molecule ICAM-1 in human cervical carcinoma cell lines.. The expression of surface antigens (major histocompatibility complex class I and class II and ICAM-1) was evaluated by fluorescence-activated cell sorter analysis in 3 human cervical carcinoma cell lines after exposure to therapeutic doses of retinoic acid. In addition, the effects on human leukocyte antigen class I messenger ribonucleic acid expression were also evaluated by Northern blot analysis after such treatment.. CaSki, SiHa, and HT-3 cervical cancer cells expressed variable levels of major histocompatibility complex class I and ICAM-1 antigens, whereas class II surface antigens were not detectable. Exposure to therapeutic doses of retinoic acid were able to significantly increase the expression of major histocompatibility complex class I and ICAM-1 antigens in all the cell lines when compared with untreated tumor cells but were not able to induce the expression of class II surface human leukocyte antigens. Northern blot analysis showed that for major histocompatibility complex class I molecules such up-regulation was the result of an increased expression at the transcriptional level of major histocompatibility complex class I messenger ribonucleic acid.. These data indicate that retinoic acid increases the expression of immunologically important surface antigens, suggesting that the efficacy of retinoic acid in the treatment of cervical cancer may be, at least in part, the result of immunologic modulation. Such findings support additional clinical research investigating the use of retinoids for the treatment of cervical cancer.

Topics: Blotting, Northern; Female; Flow Cytometry; Fluorescent Antibody Technique, Indirect; Gene Expression Regulation, Neoplastic; Histocompatibility Antigens Class I; Histocompatibility Antigens Class II; Humans; Intercellular Adhesion Molecule-1; RNA, Messenger; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Recent studies have revealed promising leads on the potential of interferons (IFNs) in combination with retinoids in solid tumor therapy. The role of IFN-alpha and retinoic acid (RA) in cervical cancer is currently under active study. Because preclinical and clinical data on IFN-beta in combination with retinoids show promising results against breast carcinoma, we analysed the anti-proliferative effect of human recombinant IFN-beta alone or in combination with all-trans RA on two human squamous cervical carcinoma cell (SCC) lines (ME180 and SiHa). The two cell lines differ in their sensitivity to the anti-proliferative effects of the different agents and their combination: i) both cell lines were more responsive to IFN-beta than to IFN-alpha2b; ii) combined treatment with RA increases the growth inhibitory effect of the single agents in ME180, but not in SiHa; iii) the antiproliferative effect correlates with the induction of apoptosis. We suggest as a possible mechanisms of action that interferon regulatory factor-1 (IRF-1), a transcription factor which belongs to the IFN machinery, and the cyclin-dependent kinase inhibitor (CDKi) p21 can be involved in cellular growth inhibition and in the induction of apoptosis. These results support the use of IFN-beta in further clinical investigation possibly in combination with retinoids.

Topics: 2',5'-Oligoadenylate Synthetase; Apoptosis; Carcinoma, Squamous Cell; Cell Division; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinases; Cyclins; DNA Fragmentation; DNA-Binding Proteins; Drug Therapy, Combination; Enzyme Inhibitors; Female; Gene Expression; Humans; Interferon alpha-2; Interferon Regulatory Factor-1; Interferon-alpha; Interferon-beta; Phosphoproteins; Recombinant Proteins; RNA, Messenger; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Topics: Antineoplastic Agents; Apoptosis; Carcinoma; Cell Survival; Cyclin-Dependent Kinase Inhibitor p21; Cyclins; Drug Synergism; Female; Gene Expression; Humans; Retinoids; Tretinoin; Tumor Cells, Cultured; Tumor Suppressor Protein p53; Uterine Cervical Neoplasms

The ability of all-trans retinoic acid (atRA), interferon-alpha2a (IFN-alpha2a) or a combination thereof to modulate the growth of three human cervical carcinoma cell lines (ME180, MS751 and CaSki) and the relationship between responsiveness and the expression of cytosolic retinoid-binding proteins (CRBP and CRABP II), nuclear RA receptors (RAR-alpha, -beta and -gamma) and retinoid X receptor (RXR alpha) were investigated. atRA induced an antiproliferative effect on two of the cell lines (ME180 > MS751), whereas CaSki was much less responsive. An additive growth inhibition on the latter two cell lines was achieved with the combined treatment of atRA and IFN-alpha2a. Receptor expression appeared to be unrelated to growth inhibition in these cell lines in so far as atRA exerted minimal effect on the growth of CaSki, although these cells expressed four of these nuclear receptors. However, mRNA for CRABP II was not demonstrable in CaSki, in contrast to the other two atRA responsive cell lines, as evaluated with RT-PCR and ethidium bromide staining. Treatment with atRA or IFN-alpha2a did not induce any change in mRNA for the nuclear retinoid receptors or cellular retinoid binding proteins after 3 or 6 days of treatment.

Topics: Antineoplastic Agents; Carcinoma; Cell Division; Cell Line; Drug Synergism; Female; Humans; Interferon alpha-2; Interferon-alpha; Oligonucleotide Probes; Polymerase Chain Reaction; Receptors, Retinoic Acid; Recombinant Proteins; Tretinoin; Uterine Cervical Neoplasms

Retinoids have been demonstrated to inhibit epithelial cell growth and differentiation. We examined the anti-proliferative effects of retinoic acid (RA) in an HPV positive and negative cervical carcinoma cell line. Our findings indicate that HPV-negative C33A cervical carcinoma cells are more sensitive to the growth inhibitory activity of retinoic acid (RA) than are HPV-positive CaSki cervical carcinoma cells. However, conditioned medium from RA-treated C33A cells displayed strong growth inhibitory activity in both C33A and CaSki cells. Since RA has been shown to modulate the expression of insulin-like growth factor binding proteins (IGFBPs) in many cells, we examined RA regulated expression of IGFBPs in medium isolated from RA treated C33A cells. IGFBP-5 was detectable in medium from C33A cells exposed to RA, and addition of purified exogenous IGFBP-5 resulted in growth inhibition of C33A cells. These results indicate that RA exerts it's anti-neoplastic effect in HPV negative cervical carcinoma cells via the overproduction of IGFBP-5.

Topics: Antineoplastic Agents; Carcinoma; Cell Division; Female; Growth Inhibitors; Humans; Insulin-Like Growth Factor Binding Protein 5; Papillomaviridae; Papillomavirus Infections; Tretinoin; Tumor Cells, Cultured; Tumor Virus Infections; Uterine Cervical Neoplasms

Retinoids inhibit the growth and reverse aberrant differentiation of squamous cell carcinoma (SCC) cells in vitro. To investigate the potential mechanisms of antitumor activity of retinoids in vivo, we used the cervical SCC cell line ME-180 as a s.c. tumor xenograft in athymic nude mice. After s.c. injection, tumor cells were allowed to form visible tumors and antitumor activity of all-trans-retinoic acid (tRA) was studied. tRA was administered daily for a 1-week or a 2-week period at 60 mg/kg/day. Tumor specimens were then analyzed using immunohistochemical staining for the number of blood vessels and apoptotic cells and for proliferating cell nuclear antigen expression. Furthermore, we studied the effect of the tRA treatment on the expression of a binding protein for fibroblast growth factors (BP; Gen-Bank accession no. M60047) that is a candidate angiogenesis modulator in SCC (F. Czubayko et al., J. Biol. Chem., 269: 28243-28248, 1994). We found that in vivo tRA treatment reduces BP expression in SCC xenografts, inhibits their angiogenesis, induces apoptosis of the tumor cells, and leads to a decrease of the tumor growth rate. We speculate that the tRA down-regulation of BP is responsible for the reduction of angiogenesis.

Topics: Animals; Antineoplastic Agents; Apoptosis; Carcinoma, Squamous Cell; Cell Differentiation; Drug Screening Assays, Antitumor; Female; Humans; Keratinocytes; Mice; Mice, Nude; Neoplasm Transplantation; Neovascularization, Pathologic; Proliferating Cell Nuclear Antigen; RNA, Messenger; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

The potential of retinoic acid as a radiosensitizer was investigated using SiHa and CC-1 human uterine cervical carcinoma cell lines, representative of high- and low-grade lesions, respectively. SiHa was significantly (P < 0.05) radiosensitized, whereas CC-1 was not. Although 48 h of treatment with 5 microM 13-cis-retinoic acid prior to irradiation was sufficient to induce radiosensitization, continuation of treatment after irradiation significantly increased the effect (P < 0.05). Three hypotheses were tested to explain the different responses of the two lines. One hypothesis was that SiHa is more sensitive to retinoic acid than CC-1. Measurement of growth revealed that SiHa was more sensitive to growth inhibition by retinoic acid than CC-1. The second hypothesis was that retinoic acid increases the proportion of G1-phase cells in SiHa but not in CC-1. This was found not to be true, because a retinoic acid treatment schedule that induced radiosensitization did not alter cell cycle distribution profiles in the absence of radiation. The third hypothesis was that retinoic acid alters the cell cycle response of SiHa but not CC-1 to radiation. Postirradiation cell cycle profiles revealed that retinoic acid increased G1 delay in SiHa, whereas CC-1 exhibited no significant G1 delay. Both lines exhibited G2 delays that were unaffected by retinoic acid. In conclusion, radiosensitization of SiHa but not CC-1 may be explained by different sensitivities to retinoic acid and differences in postirradiation cell cycle responses. Radiosensitization at radiation doses used clinically was observed when retinoic acid was administered both before and after irradiation.

Topics: Cell Cycle; Cell Division; Cell Survival; Cobalt Radioisotopes; Dose-Response Relationship, Radiation; Female; Humans; Radiation Tolerance; Radiation-Sensitizing Agents; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Retinoic acid receptor (RAR) active retinoids have proven therapeutically useful for treating certain cancers and dermatological diseases. Herein, we describe the discovery of two new RAR active trienoic acid retinoids, (2E,4E,6E)-7-(3,5-di-tert-butylphenyl)-3-methylocta-2, 4,6-trienoic acid (10a, ALRT1550) and (2E,4E,6Z)-7-(3,5-di-tert-butylphenyl)-3-methylocta-2, 4,6-trienoic acid (10b, LG100567). ALRT1550 is a RAR selective retinoid which exhibits exceptional potency in both competitive binding and cotransfection assays. Moreover, it is the most potent antiproliferative retinoid described to date and thus has implications for the treatment of certain cancers. LG100567 is a potent panagonist which activates both RARs and retinoid X receptors.

Topics: Antineoplastic Agents; Cell Division; Drug Screening Assays, Antitumor; Female; Humans; Molecular Structure; Receptors, Retinoic Acid; Retinoid X Receptors; Retinoids; Thymidine; Transcription Factors; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Human papillomaviruses (HPVs) and cigarette smoking are epidemiologically associated with cervical cancer. We recently found that HEN-16 and HEN-16-2 HPV type 16-immortalized endocervical cells form tumors after treatment with cigarette smoke condensate and derived 2 tumor cell line cultures, HEN-16T and HEN-16-2T, respectively. Here, we examine the molecular pathologic effect of tumorigenesis. HEN-16T and HEN-16-2T exhibit unchanged status and expression of integrated HPV 16 DNA. However, the expression of the cytokeratin CK7 and CK13 endocervical cell markers is more homogeneous in monolayer and organotypic raft cultures after tumorigenesis. For the effect of retinoic acid on monolayers for growth inhibition, HEN-16T were significantly less sensitive than the normal and immortalized non-tumorigenic cells. HEN-16-2T were completely resistant. Moreover, the rafts from both tumorigenic cell line cultures were resistant to retinoic acid and continued to display thick rafts and homogeneous severe dysplasia/carcinoma in situ. In contrast, the non-malignant HEN-16 and HEN-16-2 rafts were thinner, and treatment with retinoic acid blocked the formation of severe dysplasia, reconstructing an epithelium resembling that of the normal endocervix. Our results support the significance of non-viral factors in the mechanism by which cigarette smoking induces tumorigenesis in the late stages of HPV-initiated progression to cervical cancer. Importantly, our data indicate that the sensitivity to retinoic acid of the HPV-containing endocervical cells is lost following tumorigenesis in vitro and possibly in women.

Topics: Cell Division; Cell Line, Transformed; Cell Transformation, Neoplastic; Cell Transformation, Viral; DNA, Viral; Drug Resistance; Female; Humans; Keratins; Keratolytic Agents; Papillomaviridae; Tretinoin; Uterine Cervical Neoplasms

The purpose of this study was to determine whether all-trans-retinoic acid (RA) can inhibit the growth of cervical neoplastic cells by inducing differentiation or by increasing the secretion of transforming growth factor-beta (TGF-beta). Normal and HPV DNA-positive cervical cells (2 cell lines derived from cervical intraepithelial neoplasia (CIN), 2 HPV DNA-transfected cell lines, and 6 cervical carcinoma cell lines) were treated with RA (1 to 1000 nM), and both total viable cell count and [3H]thymidine incorporation were used to evaluate proliferation. In vitro differentiation was evaluated in organotypic (collagen gel raft) cultures with hematoxylin/eosin staining, and using specific immunostaining for fillagrin and cytokeratin 10. TGF-beta 1 and TGF-beta 2 secretion were measured with specific SELISAs. One-way analysis of variance and t tests were performed. RA causes a dose-dependent (P<0.05) growth arrest of comparable magnitude in normal ectocervical cells, in HPV DNA-transfected cell lines, in CIN-derived cell lines, and in four of six carcinoma cell lines. Endocervical cells and two carcinoma cell lines are unaffected. In vitro differentiation is decreased in CIN cells and is unchanged in carcinomas treated with RA as compared to control. Secretion of either TGF-beta 1 or TGF-beta 2 is significantly increased (P<0.05) in response to RA, both in RA-sensitive and in RA-resistant cells. RA induces growth inhibition in cervical neoplastic cell lines, including cervical carcinoma cells. This does not appear to be the result of increased differentiation or of increased TGF-beta secretion.

Topics: Cell Count; Cell Differentiation; Cell Division; Cell Line, Transformed; Cervix Uteri; DNA, Viral; Dose-Response Relationship, Drug; Female; Humans; Papillomaviridae; Reference Values; Transfection; Transforming Growth Factor beta; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Interferon and retinoic acid are active agents for the treatment of cervical cancer, but their mechanisms of action are unclear. Results of [3H]thymidine uptake assays showed that exposure to pharmacologic concentrations of interferon-alpha (IFN-alpha) and all-trans-retinoic acid (RA) for 72 hr inhibited growth of the cervical cancer cell lines ME-180, 283, SiHa, C33-A, 621, CaSki, HeLa, and B132, CaSki and SiHa cells continuously exposed to IFN-alpha or RA or both for 9 days developed resistance to growth inhibition, and growth resumed at a rate comparable to control after removal of agents. Similar assays showed no significant difference in effects of RA and its cis isomer. Assays for lactate dehydrogenase release revealed no significant lysis of any cell line following exposure to IFN-alpha, RA, or their combination. In organotypic culture, cells grew in a pattern histologically similar to carcinoma in situ, and exposure to IFN-alpha and RA for 14 days yielded no change in this pattern. Immunohistochemical analysis showed no change in cytokeratin expression by cells in organotypic or monolayer culture. The major in vitro effect of IFN-alpha and RA on cervical cancer cell lines appears to be reversible inhibition of proliferation.

Topics: Antineoplastic Combined Chemotherapy Protocols; Cell Division; Female; Humans; Immunohistochemistry; Interferon-alpha; Time Factors; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

We have created fusion proteins between Fas and the ligand-binding domain of the estrogen or retinoic acid receptor. Murine fibrosarcoma L929 cells and human cervical carcinoma HeLa cells expressing the fusion proteins demonstrated apoptotic phenotypes in a tightly estrogen- or retinoic acid-dependent manner in vitro. Moreover, the fusion protein-expressing L929 cells transplanted into nude mice were also killed through apoptosis after injection of an estrogen agonist. This represents a novel system, "cell targeting," that can eliminate cells not only in vitro but also in vivo through the activation of a natural suicide machinery, i.e., apoptosis, by currently used hormones. This system implies wide applications not only in developmental biology and neurobiology but also in medicine, especially for cancer gene therapy.

Topics: Amino Acid Sequence; Animals; Apoptosis; Cell Line; Cell Nucleus; Chlorocebus aethiops; Estradiol; Estriol; fas Receptor; Female; Fibrosarcoma; HeLa Cells; Humans; Male; Mice; Mice, Nude; Molecular Sequence Data; Receptors, Estrogen; Recombinant Fusion Proteins; Sequence Tagged Sites; Tamoxifen; Transfection; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Topics: Adult; Antineoplastic Agents; Carcinoma in Situ; Combined Modality Therapy; Female; HIV Infections; Humans; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms; Vaginal Neoplasms; Vulvar Neoplasms

Topics: Alitretinoin; Animals; Anticarcinogenic Agents; Antineoplastic Agents; Breast Neoplasms; Clinical Trials, Phase I as Topic; Clinical Trials, Phase II as Topic; Drug Approval; Female; Humans; Neoplasms, Experimental; Rats; Research Design; Tretinoin; Uterine Cervical Neoplasms

To determine the effect of retinoic acid on the development of severe dysplasia or carcinoma in situ from endocervical cells containing human papillomavirus (HPV) type 16.. Two independent lines of HPV 16-immortalized endocervical cells were reconstructed into two squamous epithelial tissues using the organotypic raft culture system to examine the differentiated phenotype. The effect of retinoic acid on dysplastic morphology of differentiation of the epithelia was examined by light microscopy of stained sections and electron microscopy. The endocervical cell type cytokeratin expression pattern was determined by indirect immunofluorescence using specific monoclonal antibodies. Ribonucleic acid expression of the HPV 16 E7 oncogene was examined by in situ hybridization.. Untreated HPV 16-immortalized endocervical cells were reconstructed into squamous dysplastic lesions resembling carcinoma in situ observed in women. Retinoic acid-treated rafts formed epithelia composed of two to three cell layers of columnar-like cells resembling simple epithelium of the endocervix. Electron microscopy and cytokeratin expression patterns confirmed the histology of a differentiated endocervical phenotype after treatment with retinoic acid. Expression of HPV 16 E7 was modestly lower in treated epithelia, preferentially in basal cells.. Retinoic acid prevents the histology and cytokeratin differentiation markers of carcinoma in situ of HPV 16-immortalized endocervical cells. Because the epithelia closely mimic HPV 16-containing severe dysplasias and native endocervical epithelium in women, this immortalized endocervical cell-raft system may be useful as a model to assess the efficacy of agents such as retinoic acid for preventing progression of these lesions to malignant cervical carcinoma.

Topics: Carcinoma in Situ; Cell Line, Transformed; Cell Transformation, Neoplastic; Cells, Cultured; Cervix Uteri; Female; Humans; Keratins; Microscopy, Electron; Papillomaviridae; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

The in vivo and in vitro antitumor effectiveness of IFNs is well documented. Their combination with differentiating agents, such as retinoic acid, has been demonstrated to be a promising therapy for patients with advanced squamous cell cancer of the skin and the cervix. However, the mechanisms that mediate these antitumor responses are not yet known. We studied the epidermoid cell line ME 180 derived from human cervical carcinoma to test its responsiveness to IFN-alpha-2b (INTRON A) and all-trans-retinoic acid (RA). Both agents have demonstrated ability to inhibit the growth of ME 180 cells in a dose- and time-dependent manner. The antiproliferative effect was further increased by the treatment with IFN-alpha-2b and RA combined. In accordance with this result, we found that the combination of the two agents has the effect of increasing the expression of the 2-5A synthetase gene, which is thought to play a key role in antigrowth responses to IFNs. At increased levels of 2-5A synthetase mRNA corresponds a significant increase in 2-5A synthetase activity. Although RA per se has no effect on the 2-5A synthetase expression, when it is combined with IFN-alpha-2b it appears to be able to potentiate the IFN-induced 2-5A synthetase expression. Moreover, the combination of IFN-alpha-2b and RA produces a similar effect also on the expression of the HLA-A2 gene, which has been shown to be induced in ME 180 cells both by IFN-alpha-2b and RA alone. In view of the possible mechanisms of action of the two agents, it is interesting to note that their combination increases, although transiently, the expression of IRF1, which codes for a transcription factor that regulates IFN gene expression and is thought to be involved in the regulation of IFN-induced effects and in mediating cell death or apoptosis.

Topics: 2',5'-Oligoadenylate Synthetase; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Squamous Cell; Cell Division; DNA-Binding Proteins; Drug Synergism; Female; Gene Expression; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; Interferon alpha-2; Interferon Regulatory Factor-1; Interferon-alpha; Phosphoproteins; Receptors, Retinoic Acid; Recombinant Proteins; RNA, Messenger; Stimulation, Chemical; Transcription Factors; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Retinoids (vitamin A and its natural and synthetic derivatives) have shown potential as chemopreventive agents, and diets poor in vitamin A and/or its precursor beta-carotene have been linked to an increased risk of cancer at several sites including the cervix. Human papillomavirus (HPV) plays an important role in the etiology of cervical cancer. We have developed an in vitro model of cancer progression using human keratinocytes (HKc) immortalized by HPV16 DNA (HKc/HPV16). Although immortal, early passage HKc/HPV16, like normal HKc, require epidermal growth factor (EGF) and bovine pituitary extract (BPE) for proliferation and undergo terminal differentiation in response to serum and calcium. However, following prolonged culture, growth factor independent HKc/HPV16 lines that no longer require EGF and BPE can be selected (HKc/GFI). Further selection of HKc/GFI produces lines that are resistant to serum- and calcium- induced terminal differentiation (HKc/DR). HKc/DR, but not early passage HKc/HPV16, are susceptible to malignant conversion following transfection with viral Harvey ras or Herpes simplex virus type II DNA. We have investigated the sensitivity of low to high passage HKc/HPV16 and HKc/GFI to growth control by all-trans-retinoic acid (RA, an active metabolite of vitamin A). Early passage HKc/HPV16 are very sensitive to growth inhibition by RA, and in these cells RA decreases the expression of the HPV16 oncogenes E6 and E7. However, as the cells progress in culture they lose their sensitivity to RA. Growth inhibition by RA may be mediated through the cytokine transforming growth factor-beta (TGF-beta), a potent inhibitor of epithelial cell proliferation. RA treatment of HKc/HPV16 and HKc/GFI results in a dose-and time-dependent induction (maximal of 3-fold) in secreted levels of TGF-beta. Also, Northern blot analysis of mRNA isolated from HKc/HPV16 demonstrated that RA treatment induced TGF-beta 1 and TGF-beta 2 expression about 3- and 50-fold, respectively. We next studied the effect of TGF-beta 1 and TGF-beta 2 on the proliferation of early to late passage HKc/HPVa6, HKc/GFI and HKc/DR. While early passage HKc/HPV16 were as sensitive as normal HKc to growth inhibition by TGF-beta 1 and TGF-beta 2, the cells became increasingly resistant to TGF-beta during in vitro progression, with the proliferation of HKc/DR being virtually unaffected by TGF-beta 1 or TGF-beta 2 treatment. Overall, loss of growth inhibition by RA parallels loss of TGF-beta sensi

Topics: Cell Division; Cell Line, Transformed; Cell Transformation, Neoplastic; Cell Transformation, Viral; Female; Humans; Keratinocytes; Male; Models, Biological; Papillomaviridae; RNA, Messenger; Transforming Growth Factor beta; Tretinoin; Uterine Cervical Neoplasms

Retinoic acid receptors and retinoid X receptors form heterodimers, bind to retinoic acid response elements, and transactivate the transcription of retinoid-responsive genes. Two synthetic retinoids [4-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-anthracenyl)benzoic acid (TTAB) and 6-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)-2-naphthale n ecarboxylic acid (TTNN)], which preferentially bind retinoic acid receptors, inhibited the proliferation of cervical carcinoma ME180 cells by 50% at 0.2 nM and 0.2 microM, respectively. In contrast, two other retinoids [2-(4-carboxyphenyl)-2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2- naphthalenyl)-1,3-dithiane (SR11203) and 4-(2-methyl-1-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2- naphthalenyl)propenyl)benzoic acid (SR11217)], which preferentially bind retinoic X receptors, inhibited growth by only 12 and 18% at 1 microM, respectively. The combination of suboptimal concentrations of TTAB (0.1 nM) or TTNN (10 nM) with each of the retinoic X receptor-selective retinoids at 1 microM showed more than additive effects on cell proliferation, especially with SR11217. Further increases in proliferation inhibition were observed when IFN-alpha (100 units/ml) was added to these retinoid combinations. Activation of transcription of a reporter gene linked 3' to the retinoic acid receptor beta retinoic acid response element in transiently transfected cells also exhibited additive effects when the cells were treated with combinations of TTAB or TTNN with SR11217. This additive activation of transcription may be the reason why the combination of retinoids is more effective than each retinoid alone. The results also suggest that the use of combinations of retinoids and IFN-alpha may lead to enhanced antitumor effects.

Topics: Cell Division; Chloramphenicol O-Acetyltransferase; Drug Screening Assays, Antitumor; Drug Synergism; Female; Humans; Interferon-alpha; Receptors, Retinoic Acid; Retinoids; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

The differentiation-inducing activity of tanshinone (TAN) and all-trans-retinoic acid (RA) was studied in vitro on a human cervical carcinoma cell line, ME180. The tumor cells were treated with TAN or RA in DMSO (final concentration 0.02%, V/V) on 4 successive days. Cells treated with the same concentration of DMSO alone served as control. Morphologic studies with light and transmission electron microscopy showed that the cells treated with both TAN and RA became well-differentiated. The cell growth, (as revealed by cell counting and 3H-TdR incorporation) was inhibited and the tumorigenicity in nude mice was reduced. No significant difference was observed between the cells treated with TAN and RA.

Topics: Abietanes; Animals; Antineoplastic Agents; Antineoplastic Agents, Phytogenic; Cell Transformation, Neoplastic; Female; Humans; Mice; Mice, Nude; Phenanthrenes; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

The effects of retinoids including all-trans-retinoic acid (ATRA), 13-CIS-RETINOIC ACID (13CRA), and N-(4-hydroxyphenyl)retinamide (4-HPR) on several cervical carcinoma cell lines in culture were investigated as a prelude to investigating the mechanisms underlying the chemopreventive potential of retinoids in cervical cancer. We found that when used at a concentration of 1 microM, 13CRA and ATRA inhibited the proliferation of three cell lines (ME-180 [HPV 68], SiHa [HPV 18], and HT-3 [HPV-]) by about 80% after a seven-day treatment. Three other cell lines (MS-751 [HPV 18], HeLa [HPV 18], C-33A [HPV-]) were moderately inhibited (30-48%), and two (C-4 II [HPV 18], CaSki [HPV 16]) responded poorly (< 25% inhibition). 4-HPR failed to inhibit the growth of any of these cell lines when used at 1 microM; however, when used at 5 or 10 microM, it induced apoptosis as evidenced by DNA fragmentation in several of the cell lines and was more potent in this effect than 10 microM ATRA. Retinoids that induce apoptosis in malignant cells may be able to exert similar effects on premalignant cells. Such retinoids would be expected to exhibit greater potency as chemopreventive agents than retinoids that exert only cytostatic effects.

Topics: Anticarcinogenic Agents; Apoptosis; Cell Division; DNA Damage; Drug Evaluation, Preclinical; Female; Fenretinide; Humans; Isotretinoin; Retinoids; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

The purpose of the present study was to establish culture conditions for human uterine cervical epithelial cells on permeable support and to determine how it affects cervical cell differentiation. Human ectocervical epithelial cells (hECE), HPV-16 immortalized hECE cells (ECE16-1) and Caski cells were grown on collagen-coated filters. Culture conditions, density of cells in culture and expression of epithelial and cervical-cell phenotypic markers were determined and compared in cells grown on filter and on solid support. Compared with the latter, cultures on filter had a higher cell density, hECE cells stratified to 5-12 cell layers compared to 1-3 on solid support, and cells of all three types expressed intercellular tight junctions. The cytokeratin profiles revealed differences between the three cell types as well as differences within the same cell species when grown on filter, compared to solid support. Of particular importance was the finding of a higher expression of K-13 in hECE grown on filter compared to solid support; K-13 is a marker of ectocervical cell differentiation. The cytokeratin profiles of the cultured hECE, ECE16-1 and Caski cells resembled those of ectocervical, squamous metaplastic and endocervical epithelia, respectively. hECE and ECE16-1 expressed involucrin protein, the level of which in both was higher in cells grown on filter compared to solid support. Polarization of the cultures was determined by morphology (stratification of hECE cells, expression of pseudomicrovilli in the apical cell membrane), selective apical vs. basolateral secretion of [35S]methionine- and [35S]cysteine-, [3H]fucose- and [14C]glucosamine-labeled molecules, and positive short-circuit current (Isc) under voltage-clamp conditions. Confluency of the cultures was determined by measuring transepithelial unidirectional fluxes of inert molecules with different molecular weights (MWs) through the paracellular pathway, and by measuring transepithelial conductance. The results indicated transepithelial permeability of 7-22.10(-6) cm.sec-1, which was 5-100 fold smaller compared to blank inserts, with a cut-off MW of 40-70 kDa for hECE and Caski cells. Transepithelial conductance ranged 18.5 to 51.5 mS.cm-2, indicating a leaky but confluent epithelia. Collectively the results indicate the epithelial nature of the cells and their improved differentiation when grown on filter support; hECE is a model for ectocervical epithelium while ECE16-1 and Caski express phenoty

Topics: Carcinoma, Squamous Cell; Cell Differentiation; Cell Line, Transformed; Cell Polarity; Cells, Cultured; Ceramics; Cervix Uteri; Collagen; Culture Techniques; Epithelial Cells; Epithelium; Female; Humans; Hydrocortisone; Keratins; Permeability; Polytetrafluoroethylene; Protein Precursors; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms; Vimentin

Topics: Antineoplastic Agents; Female; Humans; Remission Induction; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

In the present study, we examine the effects of all-trans-retinoic acid (RA) and interferons-alpha and -gamma (IFN-alpha and IFN-gamma) on the growth of HPV16-immortalized cell lines, ECE16-1 and CaSki. Treating proliferating ECE16-1 cells with RA causes a concentration-dependent decrease in cell number. At 1 microM RA, cell growth is suppressed by 65% and the level of mRNA encoding cytokeratin K5, a biochemical marker of retinoid action, is also suppressed. In contrast, the level of transcript encoding the HPV16 oncogenes, E6 and E7, is reduced by only 5 to 10%. IFN-alpha at 1000 IU/ml or IFN-gamma at 200 IU/ml suppresses growth by 70%. This growth suppression by IFN-gamma is correlated with a > 90% reduction in E6/E7 mRNA levels. Additional growth suppression is observed upon simultaneous treatment with retinoid and interferon. Optimal suppression is observed in the presence of 200 IU/ml IFN-gamma and 1 microM RA. The rank order of effectiveness is IFN-gamma/RA > IFN-alpha/RA = IFN-gamma > RA > IFN-alpha. In contrast to the suppression of ECE16-1 cell growth, RA causes a concentration-dependent increase in CaSki cell number (50-60%) which is optimal at 1 microM RA. Cytokeratin K5 mRNA levels are markedly suppressed, and E6/E7 mRNA levels increased by 5% under these conditions. IFN-alpha at 1000 IU/ml or IFN-gamma at 200 IU/ml decreases CaSki cell growth by 20 and 45%, respectively, and 200 IU/ml of IFN-gamma reduce E6/E7 expression to undetectable levels. Addition of RA (1 microM) partially counters the IFN-dependent suppression of growth and E6/E7 mRNA levels. Our results suggest that retinoid-dependent changes in human papillomavirus-immortalized cervical cell proliferation are not always correlated with changes in E6/E7 transcript levels.

Topics: Animals; Cell Division; Cell Line; Cell Line, Transformed; Cell Transformation, Viral; Cervix Uteri; Dose-Response Relationship, Drug; Epidermal Growth Factor; Epithelial Cells; Epithelium; Female; Gene Expression; Humans; Interferon-alpha; Interferon-gamma; Kinetics; Mice; Mice, Nude; Oncogenes; Papillomaviridae; RNA, Messenger; Transcription, Genetic; Transplantation, Heterologous; Tretinoin; Uterine Cervical Neoplasms

We describe an animal model to induce the histogenesis of squamous metaplasia of the cervical columnar epithelium, a condition usually preceding cervical neoplasia. This model is based on dietary retinoid depletion in female mice. Control sibling mice fed the same diet but with all-trans-retinoic acid (at 3 micrograms/g diet) showed the normal endocervical epithelial and glandular columnar morphology, typical of a simple epithelium without subcolumnar reserve cells. The stratified squamous ectocervical epithelium of these mice fed all-trans retinoic acid showed intense immunohistochemical staining in basal and suprabasal cells with mono-specific antibodies against keratins K5, K14, K6, K13, and, suprabasally, with antibodies specific for K1 and K10. At the squamocolumnar junction, the adjacent columnar epithelium (termed "suprajunctional") did not show staining for K5, K14, K6, K13, K1, and K10 but specifically stained for keratin K8, typical of simple epithelia and absent from the adjacent ectocervical squamous stratified lining (termed "subjunctional"), in striking contrast. Sections of the squamocolumnar junction from mice kept on the vitamin A-deficient diet for 10 weeks showed suprajunctional isolated patches of reserve cells, proximal and distal to the junction. These cells were detected prior to any symptoms of vitamin A deficiency, such as loss of body weight or respiratory discomfort. The subcolumnar reserve cells induced by vitamin A deficiency displayed positive staining for K5 and K14. As deficiency became severe, the reserve cells occupied the entirety of the suprajunctional basement membrane. This epithelium eventually became stratified and squamous metaplastic, the squamocolumnar junction was no longer discernible, and the entire endocervical epithelium and the endometrial glands lost K8 positivity, while acquiring K5, K14, K6, K13, K1, and K10 keratins typical of the ectocervix under normal conditions of vitamin A nutriture. Vitamin A deficiency also altered keratin expression and localization in squamous subjunctional epithelium. In situ hybridization studies for K1 and K5 mRNA showed their major site of expression at the basal (K5) and immediately suprabasal (K1) cell layers. The localization of both K5 and K1 proteins in these same cell layers, and above, is consistent with transcriptional regulation of these keratins. Early vitamin A deficiency caused the appearance of single subcolumnar reserve cells expressing K5 mRNA. After these ce

Topics: Animals; Carcinoma, Squamous Cell; Cervix Uteri; Diet; Disease Models, Animal; Epithelium; Female; Immunohistochemistry; In Situ Hybridization; Keratins; Metaplasia; Mice; Mice, Inbred BALB C; Mice, Nude; Phenotype; Precancerous Conditions; Retinoids; RNA, Messenger; Tretinoin; Uterine Cervical Neoplasms; Vitamin A Deficiency

The effect of drugs on the release of CA125 antigen and the binding of anti-CA125 monoclonal antibody (MoAb) to malignant cells was evaluated in vitro. TMCC-1, uterine cervical adenocarcinoma cells, were exposed to dexamethasone (DEX), sodium n-butyrate (NaB), dibutyryl cyclic AMP (dbcAMP), retinoic acid (RA), calcitriol (VD3), and interferon-gamma (IFN-gamma). NaB, RA and VD3 increased CA125 release per cell and 125I-labeled anti-CA125 MoAb binding to the cells. DEX also increased the 125I-labeled anti-CA125 MoAb binding to the cells, and CA125 antigen release per cell was also slightly increased. IFN-gamma suppressed both CA125 release and 125I-labeled MoAb binding. A combination of DEX, VD3 and RA and increased the binding of MoAb to TMCC-1 cells, but the amount of bound MoAb was not significantly different from that obtained by single drug treatment. DbcAMP had no significant effect on enhancing MoAb binding. Drugs can increase the binding of anti-CA125 MoAb to malignant cells and they may be applied to increase the tumor uptake of radiolabeled MoAbs in vivo.

Topics: Adenocarcinoma; Antigens, Tumor-Associated, Carbohydrate; Bucladesine; Butyrates; Butyric Acid; Calcitriol; Dexamethasone; Female; Humans; In Vitro Techniques; Interferon-gamma; Tretinoin; Uterine Cervical Neoplasms

Human papillomavirus type 18 (HPV18) belongs to the group of genital papillomaviruses involved in the development of cervical carcinomas. Since retinoic acid (RA) is a key regulator of epithelial cell differentiation and a growth inhibitor in vitro of HPV18-positive HeLa cervical carcinoma cells, we have used HeLa and HeLa hybrid cells in order to analyse the effects of RA on expression of the HPV18 E6 and E7 oncogenes and of the cellular RA receptor genes RAR-beta and -gamma. We show here that RA down-regulates HPV18 mRNA levels apparently due to transcriptional repression. Transient cotransfection assays indicated that RARs negatively regulate the HPV18 upstream regulatory region and that the central enhancer can confer RA-dependent repression on a heterologous promoter. RA treatment resulted in induction of RAR-beta mRNA levels in non-tumorigenic HeLa hybrid cells, but not in tumorigenic hybrid segregants nor in HeLa cells. No alterations of the RAR-beta gene or of the HeLa RAR-beta promoter could be revealed by Southern and DNA sequence analysis, respectively. As determined by transient transfection assays, however, the RAR-beta control region was activated by RA more strongly in non-tumorigenic hybrid cells than in HeLa cells, thus indicating differences in trans-acting regulatory factors. Our data suggest that the RARs are potential negative regulators of HPV18 E6 and E7 gene expression, and that dysregulation of the RAR-beta gene either causatively contributes to or is an indicator of tumorigenicity in HeLa and HeLa hybrid cells.

Topics: Base Sequence; Carrier Proteins; Cell Division; Cell Nucleus; DNA; DNA, Neoplasm; Down-Regulation; Female; Gene Expression Regulation, Viral; HeLa Cells; Humans; Hybrid Cells; Keratinocytes; Luciferases; Molecular Sequence Data; Oligodeoxyribonucleotides; Papillomaviridae; Placenta; Polymerase Chain Reaction; Pregnancy; Receptors, Retinoic Acid; Recombinant Fusion Proteins; RNA, Messenger; Transcription, Genetic; Transfection; Tretinoin; Uterine Cervical Neoplasms

Cellular retinoic acid-binding protein (CRABP) and cellular retinol-binding protein (CRBP) were localized in biopsies of normal squamous epithelium, cervical intraepithelial neoplasia (CIN), and invasive squamous cell cancer of the cervix uteri by immunohistochemistry. In both the normal stratified squamous epithelium of the exocervix and low-grade CIN, CRABP I was present predominantly in the basal layer of the epithelium. The more superficial, differentiated cell layers lacked immunoreactive protein. In high-grade CIN (CIN2-3), the distribution of CRABP I was altered. Immunoreactive CRABP I was detected in all layers of high-grade CIN. In squamous cell carcinoma of the cervix, CRABP I was detected in cells throughout the tumor but was minimal in cells demonstrating squamous differentiation. In contrast to CRABP I, CRBP was diffusely present throughout the cervical epithelium irrespective of the state of differentiation or the presence of disease.

Topics: Adolescent; Adult; Carcinoma, Squamous Cell; Carrier Proteins; Cervix Uteri; Female; Humans; Middle Aged; Neoplasm Proteins; Receptors, Retinoic Acid; Retinol-Binding Proteins; Retinol-Binding Proteins, Cellular; Tretinoin; Uterine Cervical Neoplasms

Retinoic acid (RA) and epidermal growth factor (EGF) regulate growth and differentiation of epithelial cells. RA has both direct and indirect effects on gene expression. Direct effects result from modulation of the transcriptional activity of genes, which contain RA response elements (RARE) recognized by trans-acting nuclear RA receptors (RARs). A second indirect mechanism for the modulatory effects of RA is by the induction or repression of growth factors and growth factor receptors. There is evidence for functional interactions between RA and the EGF receptor (EGFR). RA enhances the proliferative response of cultured keratinocytes to EGF, increases the number of EGFRs on the surface of some cells, and induces EGFR promoter activity in most cells. In contrast, immunoprecipitation, Northern blot, and nuclear run-on analysis described in this paper show that RA suppresses EGFR synthesis at the transcriptional level in human epidermoid carcinoma ME180 cells. Deletion analysis of EGFR gene promoter mutants linked to the chloramphenicol acetyltransferase gene revealed the existence of a region of the promoter, -771 to -384, which is responsive to RA. Gel retardation data indicated that a cell-type nuclear protein which binds to this novel element is suppressed by RA in a dose-dependent manner. This decrease coincides with a decreased steady-state level of RAR-gamma mRNA. These data strongly suggest that the EGFR promoter is regulated by RAR-gamma, which itself is under the control of RA. Other cell-specific trans-acting factors may be involved in this regulation.

Topics: Carcinoma, Squamous Cell; Cell Differentiation; Down-Regulation; ErbB Receptors; Female; Humans; Promoter Regions, Genetic; Recombinant Fusion Proteins; Regulatory Sequences, Nucleic Acid; RNA, Messenger; Transcription, Genetic; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Human cervical cells are a primary site of papillomavirus infection and 90% of all cervical tumors are positive for human papillomavirus (HPV) DNA. Over one-half million cases of HPV-associated cervical, vulvar, and penile cancers are reported per year. Yet, in spite of the magnitude of this problem, the effects of HPV infection on cervical cell growth and differentiation are not well characterized. To study these effects we have developed a clonal cell line of HPV-16-immortalized ectocervical epithelial cells, ECE16-1. In the present study we demonstrate that under normal growth conditions the cytokeratin content of ECE16-1 cells is dramatically altered compared to normal cervical cells; the level of K5, K6, K14, K16, and K17 is reduced and the level of K7, K8, and K19 is increased. We demonstrate that this change is largely due to a difference in the response of the cells to retinoids, as growth in retinoid-free medium produces a complete normalization of cytokeratin levels. Upon addition of natural and synthetic retinoids, the levels of cytokeratins K5, K6, K14, K16, and K17 are reduced, while the levels of cytokeratins K19, K7, and K8 are increased. Cytokeratin K13 levels are only slightly altered. The level of involucrin, a precursor of the cervical cell envelope (superficial cell), is not changed by immortalization nor is it regulated by retinoids. Transglutaminase activity is also not appreciably altered by immortalization; however, ECE16-1 cells make fewer envelopes than normal ECE cells. Our results clearly indicate that natural and synthetic retinoids suppress the differentiation of HPV transformed cervical cells. In early, low grade, cervical intraepithelial neoplasia, transcription of the HPV16 E6/E7 oncogenes is confined to the suprabasal layers. Our results suggest that retinoids, because they inhibit the differentiation of HPV16 immortalized cervical cells, may reduce the extent of viral oncogene transcription and thus be useful in slowing the neoplastic process.

Topics: Cell Differentiation; Cell Transformation, Viral; Cells, Cultured; Cervix Uteri; Epithelial Cells; Epithelium; Female; Humans; Keratins; Papillomaviridae; Phenotype; Retinoids; RNA, Messenger; Transcription, Genetic; Transfection; Tretinoin; Uterine Cervical Neoplasms; Virus Replication; Vitamin A

Retinoic acid (RA) increases epidermal growth factor (EGF) receptors in many cells; in ME180 cells, a human epidermoid carcinoma, RA resulted in a dose- and time-dependent reduction of EGF binding. In RA-treated ME180 cells, binding was 41% of the control. The reduction of EGF binding was due to a decrease in the number of receptors, from 8.7 x 10(4) to 3.6 x 10(4) per cell. The difference was present 8 h after the addition of RA and was reversible 3 days after its removal. Scatchard analysis indicated that RA did not change the binding affinity of EGF (Kd = 1 nM). Also, RA did not alter the rate of EGF internalization or the down-regulation induced by exogenous EGF. Flow-cytometric analysis revealed that RA did not alter the cell cycle. Soluble cell membrane extracts were prepared in a Tris buffer with protease inhibitors, immunoprecipitated, electrophoresed, and immunoblotted with an antiserum to EGF receptors. The EGF receptor band of Mr 170,000 was decreased in RA-treated cells. These results suggest that RA reduces the synthesis of EGF receptors in ME180 cells.

Topics: Carcinoma, Squamous Cell; Cell Division; Down-Regulation; Epidermal Growth Factor; ErbB Receptors; Female; Humans; Time Factors; Tretinoin; Tumor Cells, Cultured; Uterine Cervical Neoplasms

Twenty seven women with mild, moderate or severe cervical dysplasia proven by pathology were treated by retinamide RII suppository. Retinamide RII suppository, 10 mg QD, was given intravaginally for six months (three months as a course). Clinical examination, Papanicolaou cytology and tissue biopsy under colposcope were carried out before and after treatment. The results indicated that after the second course, 26 out of 27 patients responded; of them, precancerous lesions disappeared in 24 and even normal squamous epithelium was observed in 3. The overall response rate was 96.29% and the marked response rate was 88.89%. The general side effects were mild. There was little cervical and vaginal irritation which was well tolerated. The results of this clinical trial make available a practical base for chemoprevention of cervical cancer.

Topics: Antineoplastic Agents; Cervix Uteri; Female; Humans; Hyperplasia; Precancerous Conditions; Suppositories; Tretinoin; Uterine Cervical Neoplasms

When the human cell line NHIK 3025 was exposed to retinoic acid (1 nM; 10 microM), the cell cycle time was prolonged. Experiments using cells synchronized by mitotic selection showed that the retinoic acid induced growth delay was barely seen within the first cell cycle after exposure to 10 microM retinoic acid, whereas the next cell cycle durations were increased 30-60%. The effect was reversible as normal growth rate was restored after removal of the drug. DNA histograms indicated a prolongation of G1 of the cell cycle. We have shown earlier that glucocorticoid steroids also induce a prolongation of the cell cycle, located within G1. When the cells were exposed to the synthetic glucocorticoid, dexamethasone, in addition to retinoic acid, no additive effect was found; on the contrary, growth inhibition was less than that with retinoic acid alone. Dexamethasone from 1 nM upwards antagonized the growth inhibitory effect of retinoic acid. This glucocorticoid mediated effect seemed to be mediated via the glucocorticoid receptor, as no effect was seen when the receptor was blocked by the antagonist 17 beta hydroxy-11 beta, 4-dimethylaminophenyl-17 alpha-propynyl estra 4,9 diene-3-one [RU 38486]. The growth inhibition studies were supported by morphological observations showing that dexamethasone induced cytoskeletal alterations dominated when the cells were exposed to both drugs simultaneously. These findings might be of importance in cancer therapy where both drugs are used.

Topics: Cell Cycle; Cell Differentiation; Cell Line; Cytoskeleton; Dexamethasone; Estrenes; Female; Humans; Interphase; Microtubules; Mifepristone; Tretinoin; Uterine Cervical Neoplasms

Topics: Animals; Anisoles; Butylated Hydroxyanisole; Female; Methylcholanthrene; Mice; Tretinoin; Uterine Cervical Neoplasms

Topics: Animals; Etretinate; Female; Folic Acid; Humans; Isomerism; Isotretinoin; Neoplasms; Neoplasms, Experimental; Skin Neoplasms; Tretinoin; Uterine Cervical Neoplasms; Vitamin A; Vitamin E

Women with abnormal cytology were matched with normal control subjects for age, parity, ethnicity, and socioeconomic class and participated in a blind case-control study focused on the role of nutrition in cervical dysplasia. Sucrose gradient ultracentrifugation studies for determination of the presence and concentration of the binding proteins for retinol and retinoic acid were performed on colposcopic biopsy tissue specimens. The nutritional survey revealed statistically significant differences for vitamins A and C and beta carotene. Retinol binding protein was absent or minimally detectable and inversely related to the severity of the dysplasia. It is proposed that a double-blind clinical trial be conducted to evaluate whether retinoids may pharmacologically inhibit, arrest, or reverse cervical dysplasia.

Topics: Ascorbic Acid; beta Carotene; Carcinoma in Situ; Carotenoids; Diet; Female; Humans; Retinol-Binding Proteins; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms; Vitamin A

Blinded analyses of the concentrations of binding proteins for retinol and retinoic acid (CRABP) in homogenates of cancer and normal tissue aliquots obtained from human cervix, endometrium, ovary, breast, and lung were carried out by the sucrose gradient ultracentrifugation technique. In carcinomas of the cervix and endometrium, CRABP mean values of 50.4 and 123.2 pmol/g tissue, respectively were detected. Such concentrations represent a 3- and 4-fold increase over the mean values of CRABP in the normal cervix (16.9 pmol/g) and normal endometrium (30.8 pmol/g), respectively. In carcinomas of the ovary, the mean CRABP level was 128.6 pmol/g compared to the maximal mean value of less than or equal to 0.46 pmol/g in the normal ovary. Elevated levels of CRABP were also found in breast and lung carcinomas compared to the amounts detected in the same patient in normal tissue aliquots of the same organ. The differences between CRABP concentrations in cervical, endometrial, ovarian, and breast carcinomas and those in normal tissue are statistically significant. In contrast, cellular retinol-binding protein concentrations were reduced in the endometrial, ovarian, breast, and lung carcinomas compared to normal tissues. There were no significant differences between the log-mean concentrations of cellular retinol-binding proteins in the cytosols from tissue aliquots of carcinoma of the cervix and those in the cytosols from tissue aliquots of normal cervix.

Topics: Carcinoma; Carcinoma, Squamous Cell; Carrier Proteins; Endometrium; Female; Genital Neoplasms, Female; Humans; Neoplasm Proteins; Ovarian Neoplasms; Receptors, Retinoic Acid; Retinol-Binding Proteins; Retinol-Binding Proteins, Cellular; Tretinoin; Uterine Cervical Neoplasms; Uterine Neoplasms

Topics: Animals; Drug Evaluation, Preclinical; Female; Mice; Neoplasms, Experimental; Retinaldehyde; Tretinoin; Uterine Cervical Neoplasms; Vitamin A

Cellular retinol-binding protein (CRBP) and cellular retinoic acid-binding protein are present in the cytosol of normal human uterine cervical tissues, as detected by ultracentrifugation analysis. Both binding proteins have characteristically high specificity for their respective ligands. In sucrose gradients, both proteins sediment in the 2S region and are of similar molecular weight (M.W. approximately 14,000). In blind analyses of cervical biopsies, obtained under direct vision by colposcopy of normal women (control) or from patients histopathologically diagnosed to have dysplasias or carcinoma in situ (study group), CRBP was not detectable by sucrose gradient analysis in 78.8% of the 33 abnormal biopsies, compared to 23.5% of the 34 controls. This difference was statistically significant (p less than 0.005). In biopsies in which CRBP was detected, the mean levels were 2.76 and 0.72 pmol/mg protein in the cytosol for the control and study groups, respectively. In some subjects from each group, cellular retinoic acid-binding protein but not CRBP was detected in the biopsied tissue. The presence and role of these binding proteins in vitamin A metabolism, epithelial maturation and differentiation in cervical dysplasias, and in situ lesions remain to be investigated.

Topics: Adolescent; Adult; Cervix Uteri; Cytosol; Female; Humans; Male; Middle Aged; Molecular Weight; Neoplasm Proteins; Retinol-Binding Proteins; Retinol-Binding Proteins, Cellular; Tretinoin; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms