fenretinide and Uterine-Cervical-Neoplasms

This trial examined the use of 4-hydroxyphenyl-retinamide (4-HPR), demonstrated to be a potent inhibitor of carcinogenesis in vitro and in animal models, in patients with cervical intraepithelial neoplasia (CIN) grades 2 to 3. Quantitative pathology and chromosome 9 polysomy were used to understand the biology and quantify the clinical histopathologic changes observed.. Patients were randomized to 4-HPR or placebo for 6 months and followed for six more months. Cervical biopsies were obtained at baseline, 6 months, and 12 months; the biopsies were read blinded three times by the study pathologist. Feulgen-stained sections were also obtained and analyzed using computer-assisted image cytometry. Chromosome 9 polysomy was performed on tissue slices using in situ hybridization and measured quantitatively. Statistical analyses were carried out in S-Plus (Insightful Corporation, Seattle, WA) and R.. The interim analysis, planned for 40 patients, was carried out on 39. The 6- and 12-month analyses showed a statistically significant difference between the two study arms. When code was broken, the 4-HPR-treatment arm was found to have fared less well than placebo. Analyses of Feulgen-stained sections provided a quantitative measure of the increase of DNA content and texture features. Chromosome 9 polysomy was also measured using image analysis. The changes observed were consistent with those of cells displaying cancerous changes, indicating a lack of response.. 4-HPR is not active at 200 mg/day. The interim analysis was helpful in directing the study; and, in this case, ending it. The intermediate endpoint biomarkers of quantitative histomorphometry and chromosome 9 polysomy yielded quantitative and repeatable results consistent with the findings of the clinical pathologist.

Topics: Anticarcinogenic Agents; Antineoplastic Agents; Chromosome Aberrations; Chromosomes, Human, Pair 9; Female; Fenretinide; Humans; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Previous trials of topical trans-retinoic acid treatment of cervical intraepithelial neoplasia (CIN) grades 2 and 3 led to a statistically significant regression of CIN 2, but not CIN 3. We tested N-(4-hydroxyphenyl)retinamide (4-HPR), a promising oral retinoid that has been shown to induce apoptosis through nonretinoic receptor acid-mediated pathways, for its toxicity and efficacy against CIN 2/3.. In a blinded randomized trial, 4-HPR at 200 mg/day for 6 months (with a 3-day/month drug holiday) was compared with placebo in patients with biopsy-proven CIN-2/3 [high-grade squamous intraepithelial lesions (HGSILs)]. Patients were treated with placebo or 4-HPR for 6 months, biopsied, and then followed for an additional 6 months. At the 12-month end point, they underwent either loop excision if a histological lesion was present or a biopsy from the original area of the lesion if no lesion was present.. An interim analysis of blinded data showed a significantly worse prognosis at 12 months for one group. When the code was broken because of the poorer outcomes, we discovered that the 4-HPR treatment arm was performing more poorly than was the placebo at 6 and 12 months (25 versus 44% response rates at 6 months; 14 versus 50% at 12 months). Toxicity was not significant in either arm.. 4-HPR at 200 mg/day with a 3-day/month drug holiday is not active compared with placebo in the treatment of HGSIL. Because 4-HPR is active in the laboratory, the lack of effect in our trial may indicate that higher doses are needed in patients to achieve comparable results.

Topics: Adult; Antineoplastic Agents; Cheilitis; Cross-Over Studies; Exanthema; Female; Fenretinide; Humans; Medical Futility; Patient Compliance; Photosensitivity Disorders; Time Factors; Treatment Outcome; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Several genetic alterations have been described in cervical cancers including: human papillomavirus (HPV) E6 and E7 oncoproteins, subtle sequence changes, alterations in chromosome number, chromosome translocations, and gene amplifications. This report focuses on establishing chromosome 9 polysomy as a cervical biomarker of chromosome instability and using it in a chemoprevention trial. Chromosomal instability is a feature of most human cancers and is probably an early event in the process.. We used 37 cervical cone specimens to validate chromosome 9 polysomy as a biomarker and then tested its modulation in a randomized clinical trial of 4-hydroxyphenylretinamide (4-HPR) in 39 patients with three blinded histopathologic reviews. No confounders were identified. In the present study, immunohistocytochemical analysis of Chromosome 9 polysomy was carried out and quantitatively measured.. The Cell Index, the ratio of the number of total chromosome 9 copies to the total number of ells, increases significantly in archival samples as the cervix changes from normal to CIN to invasive cancer. In the chemoprevention trial, chromosome 9 polysomy was used as a biomarker and supported the histological analysis showing that 4-HPR impaired the natural regression response.. Chromosome 9 polysomy appears to be a marker of genetic instability that can be used in chemoprevention trials as a surrogate endpoint biomarker. In this randomized trial of 4-HPR, the chromosome 9 polysomy measurements supported the clinical histopathologic reading in a quantitative manner suggesting that 4-HPR at 200 mg/day may have been inhibiting the regression seen in the placebo arm by inducing genetic instability.

Topics: Aneuploidy; Anticarcinogenic Agents; Carcinoma, Squamous Cell; Chromosomes, Human, Pair 9; Female; Fenretinide; Humans; Placebos; Randomized Controlled Trials as Topic; Reproducibility of Results; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Topical trans-retinoic acid treatment produced a statistically significant regression of cervical intraepithelial neoplasia (CIN) 2, but not CIN 3, in trials. A promising oral retinoid, 4-hydroxyphenylretinamide (4-HPR) induced apoptosis through non-retinoic acid-mediated pathways and is being studied in National Cancer Institute phase II trials in several organ sites. We studied the effects of 4-HPR on cervical cancer cell lines and on cervical epithelial cell lines immortalized by human papillomavirus (HPV). Four immortalized cervical epithelial cell lines (in vitro models of precancerous CIN lesions) and nine cervical carcinoma cell lines were studied. The growth inhibitory effect of 4-HPR was tested in monolayer culture and in semisolid medium, and concentrations required for a 50% growth inhibition within 5 days were determined. The agent 4-HPR inhibited the growth of cervical carcinoma cell lines and HPV-immortalized cell lines in a dose- and time-dependent fashion. Doses of 5 and 10 microM were more effective than 1 microM. The C33A cell line was most sensitive to 4-HPR, and HeLa and HT3 were the least sensitive. Of the HPV-immortalized cell lines, Z132, an HPV-16 immortalized cell line, was sensitive; the HPV 18-immortalized cell lines (Z173, Z183, and TCL-1) were not, although they were sensitive when grown in colonies. The agent 4-HPR is active against several cervical cancer cells lines and HPV-immortalized cervical epithelial cell lines. These findings are consistent with data from other laboratories studying other organ systems. This study helps establish a relevant biologically active dose for clinical trials in the cervix, one corresponding to the 5- and 10-microM tissue doses.

Topics: Cell Division; Cell Line, Transformed; Cervix Uteri; Clinical Trials, Phase II as Topic; Dose-Response Relationship, Drug; Epithelial Cells; Female; Fenretinide; Humans; Kinetics; Papillomaviridae; Tumor Cells, Cultured; Uterine Cervical Neoplasms; Vitamin A

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

Topics: Antineoplastic Agents; Female; Fenretinide; Humans; Randomized Controlled Trials as Topic; Treatment Outcome; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

N-(4-Hydroxyphenyl)retinamide (4HPR) is currently used in cancer prevention and therapy trials. It is thought that its effects result from induction of apoptosis. 4HPR-induced apoptosis in human cervical carcinoma C33A cells involves enhanced generation of reactive oxygen species (ROS). In this study we explored the mechanism by which 4HPR increases ROS and induces apoptosis in these cells. 4HPR induced cytochrome c release from mitochondria to cytoplasm, activated caspase-3, and caused a membrane permeability transition (MPT). All these 4HPR's effects, as well as the induction of apoptosis, were inhibited by antioxidants, which decrease ROS. Thenoyltrifluoroacetone, a mitochondrial respiratory chain (MRC) complex II inhibitor, and carbonylcyanide m-chlorophenyl hydrazone, which uncouples electron transfer and ATP synthesis and inhibits ROS generation by MRC, inhibited 4HPR-induced ROS generation very effectively. Rotenone, an MRC complex I inhibitor was less effective and azide, an MRC complex IV inhibitor, exhibited a marginal effect. In contrast, antimycin A, an MRC complex III inhibitor, enhanced 4HPR-induced ROS generation. These findings suggest that 4HPR enhances ROS generation by affecting a target between complex II and complex III, presumably coenzyme Q. This effect is followed by release of cytochrome c, increased caspase-3 activity, induction of MPT and eventual DNA fragmentation and cell death.

Topics: Anticarcinogenic Agents; Antimycin A; Antineoplastic Agents; Antioxidants; Apoptosis; Carbonyl Cyanide m-Chlorophenyl Hydrazone; Carcinoma, Squamous Cell; Caspase 3; Caspases; Cytochrome c Group; Electron Transport; Electron Transport Complex I; Electron Transport Complex II; Electron Transport Complex III; Enzyme Inhibitors; Female; Fenretinide; Humans; Mitochondria; Multienzyme Complexes; NADH, NADPH Oxidoreductases; Neoplasm Proteins; Oxidoreductases; Reactive Oxygen Species; Rotenone; Sodium Azide; Succinate Dehydrogenase; Thenoyltrifluoroacetone; Tumor Cells, Cultured; Uncoupling Agents; Uterine Cervical Neoplasms

Topics: Anticarcinogenic Agents; Antineoplastic Agents; Apoptosis; Female; Fenretinide; Free Radicals; Humans; Neoplasms; Oxygen; Signal Transduction; Uterine Cervical Neoplasms

The inhibitory effects of N-(4-hydroxyphenyl)retinamide (4HPR) on tumorigenesis and tumor growth may result from its ability to induce apoptosis (programmed cell death). Since antioxidants inhibit 4HPR-induced apoptosis, experiments were planned to determine whether the levels of reactive oxygen species increase in cells undergoing apoptosis after exposure to 4HPR.. Cells of the human cervical carcinoma cell line C33A and normal human cervical epithelial cells were treated with 4HPR and analyzed for survival, induction of apoptosis, generation of reactive oxygen species, and expression of the apoptosis-related proteins Bcl-2 and Bax.. Treatment with 4HPR decreased C33A cell number by inducing apoptosis in a time- and dose-dependent fashion. DNA fragmentation typical of apoptosis was observed in cells exposed to 4HPR at concentrations of 3 microM or higher for 6-24 hours. The generation of reactive oxygen species was enhanced by 1.85-fold to 4.5-fold after a 1.5-hour treatment with 0.4-10 microM 4HPR. Pyrrolidine dithiocarbamate, an oxygen radical scavenger, suppressed the rate of generation of reactive oxygen species and inhibited 4HPR-induced apoptosis. 4HPR failed to modulate cellular levels of the Bcl-2 and Bax proteins. N-(4-Methoxyphenyl)retinamide, the major 4HPR metabolite, and several other retinoids that bind to nuclear retinoic acid receptors or retinoid X receptors failed to enhance the generation of reactive oxygen species and to induce apoptosis. 4HPR was much less effective in generating reactive oxygen species and in inducing apoptosis in normal human cervical epithelial cells than in C33A cervical carcinoma cells.. Enhancement of the generation of reactive oxygen species may be involved in apoptotic pathway induction by 4HPR.

Topics: Antioxidants; Apoptosis; bcl-2-Associated X Protein; Cell Survival; Cervix Uteri; DNA Fragmentation; DNA, Neoplasm; Dose-Response Relationship, Drug; Epithelium; Female; Fenretinide; Free Radicals; Gene Expression Regulation, Neoplastic; Humans; Oxygen; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-bcl-2; Pyrrolidines; Retinoids; Thiocarbamates; 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