thymosin and Breast-Neoplasms

Thymosin beta15 is a small actin-binding protein upregulated in highly metastatic rat prostate cancer cells, relative to low metastatic cells. We have previously established an important role for thymosin beta15 as a diagnostic marker in human prostate cancer, with potential as a prognostic indicator. We here review the data supporting increased thymosin beta15 expression in other cancer types, including breast, brain, and lung. Human NB thymosin beta is a beta-thymosin originally found in neuroblastoma. New data demonstrate that NB thymosin beta represents the human homolog of rat thymosin beta15; thus we suggest classification as human thymosin beta15. In addition to the previously described gene, thymosin beta15a, we report the discovery of a new isoform of human thymosin beta15, thymosin beta15b, which is transcribed from an independent gene on human chromosome X. The gene structure of thymosin beta15a and beta15b is conserved and the isoforms show 87% identity across the nucleotide sequence. Across the coding sequence the nucleotide differences are silent, resulting in identical proteins. Other thymosin family members have recently been shown to exert potent clinical effects. The functional data available for thymosin beta15, combined with the tumor expression pattern, suggest that thymosin beta15 may play an important role in tumor development and progression in addition to its value as a biomarker in prostate cancer.

Topics: Amino Acid Sequence; Animals; Brain Neoplasms; Breast Neoplasms; Endometrial Neoplasms; Female; Head and Neck Neoplasms; Humans; Male; Molecular Sequence Data; Prostatic Neoplasms; Protein Isoforms; Rats; RNA, Messenger; Sequence Alignment; Sequence Homology, Amino Acid; Thymosin

Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Humans; Immunologic Factors; Precancerous Conditions; Stomach Neoplasms; Thymosin

Topics: Adjuvants, Immunologic; Animals; Aziridines; BCG Vaccine; Breast Neoplasms; Cyclophosphamide; Dose-Response Relationship, Immunologic; Humans; Immunotherapy; Leucine; Leukemia L1210; Leukemia, Lymphoid; Levamisole; Lymphoma, Non-Hodgkin; Melanoma; Methotrexate; Mice; Neoplasms; Propionibacterium acnes; T-Lymphocytes, Regulatory; Thymosin

Thymosin beta 10 (TMSB10) has been demonstrated to be involved in the malignant process of many cancers. The purpose of this study was to determine the biological roles and clinical significance of TMSB10 in breast cancer and to identify whether TMSB10 might be used as a serum marker for the diagnosis of breast cancer.. TMSB10 expression was evaluated by immunohistochemical analysis (IHC) of 253 breast tumors and ELISA of serum from 80 patients with breast cancer. Statistical analysis was performed to explore the correlation between TMSB10 expression and clinicopathological features in breast cancer. Univariate and multivariate Cox regression analysis were performed to examine the association between TMSB10 expression and overall survival and metastatic status. In vitro and in vivo assays were performed to assess the biological roles of TMSB10 in breast cancer. Western blotting and luciferase assays were examined to identify the underlying pathway involved in the tumor-promoting role of TMSB10.. We found TMSB10 was upregulated in breast cancer cells and tissues. Univariate and multivariate analysis demonstrated that high TMSB10 expression significantly correlated with clinicopathological features, poor prognosis and distant metastases in patients with breast cancer. Overexpression of TMSB10 promotes, while silencing of TMSB10 inhibits, proliferation, invasion and migration of breast cancer cells in vitro and in vivo. Our results further reveal that TMSB10 promotes the proliferation, invasion and migration of breast cancer cells via AKT/FOXO signaling, which is antagonized by the AKT kinase inhibitor perifosine. Importantly, the expression of TMSB10 is significantly elevated in the serum of patients with breast cancer and is positively associated with clinical stages of breast cancer.. TMSB10 may hold promise as a minimally invasive serum cancer biomarker for the diagnosis of breast cancer and a potential therapeutic target which will facilitate the development of a novel therapeutic strategy against breast cancer.

Topics: Biomarkers, Tumor; Breast Neoplasms; Carcinogenesis; Cell Line, Tumor; Cell Movement; Cell Proliferation; Female; Forkhead Transcription Factors; Humans; Kaplan-Meier Estimate; Neoplasm Metastasis; Prognosis; Proportional Hazards Models; Proto-Oncogene Proteins c-akt; Signal Transduction; Thymosin

Thymosin alpha 1 (Tα1), an immunoactive peptide, has been shown to inhibit cell proliferation and induce apoptosis in human leukemia, non-small cell lung cancer, melanoma, and other human cancers. However, the response and molecular mechanism of breast cancer cells exposed to Tα1 remain unclear. PTEN, a tumor suppressor gene, is frequently mutated in a variety of human cancers. In the present study, we aimed to investigate the biological roles of PTEN in the growth inhibition of human breast cancer cells exposed to Tα1. Using wild-type and mutant PTEN-expressing cells, we found a strong correlation between PTEN status and Tα1-mediated growth inhibition of breast cancer cells. The growth inhibition effect was more pronounced in breast cancer cells in which Tα1 enhanced PTEN expression, whereas endogenous PTEN knockdown reversed the growth inhibition effect of Tα1 in breast cancer cells. Further investigation revealed that PTEN up-regulation, which was induced by Tα1, can inhibit the activation of the PI3K/Akt/mTOR signaling pathway, leading to the growth inhibition of breast cancer cells. The addition of the synergy between Tα1 and the inhibition of PI3K/Akt/mTOR activation could strongly block cell viability in PTEN down-regulated breast cancer cells. PTEN-overexpressing cells not only up-regulated Bax and cleaved caspase-3/9 and PARP expression but also down-regulated Bcl-2 compared to the treatment with Tα1 alone. Together these findings suggest that PTEN mediates Tα1-induced apoptosis through the mitochondrial death cascade and inhibition of the PI3K/Akt/mTOR signaling pathway in breast cancer cells.

Topics: Apoptosis; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Female; Humans; Mitochondria; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; PTEN Phosphohydrolase; Signal Transduction; Thymalfasin; Thymosin; TOR Serine-Threonine Kinases; Up-Regulation

Thymosin alpha 1 (Tα1) is commonly used for treating several diseases; however its usage has been limited because of poor penetration of the target tissue, such as tumor cells. In the present study, Tα1-iRGD, a peptide by conjugating Tα1 with the iRGD fragment, was evaluated its performance in MCF-7 and MDA-MB-231 human breast cancer cells. Compared with the wild-type peptide, Tα1-iRGD was more selective in binding tumor cells in the cell attachment assay. Furthermore, the MTT assay confirmed that Tα1-iRGD proved more effective in significantly inhibiting the growth of MCF-7 cells in contrast to the general inhibition displayed by Tα1. Further, conjugation of Tα1 with iRGD preserved the immunomodulatory activity of the drug by increasing the proliferation of mouse spleen lymphocytes. Further, compared with Tα1 treatment, Tα1-iRGD treatment of MCF-7 cells considerably increased the number of cells undergoing apoptosis, resulting in a dose-dependent inhibition of cancer cell growth, which was associated with a much better effect on up-regulation of the expression of BCL2-associated X protein (Bax), caspase 9, etc. More importantly, treatment with Ta1-iRGD was more efficacious than treatment with Ta1 in vivo. This study highlights the importance of iRGD on enhancement of cell penetration and tumor accumulation. In summary, our findings demonstrate that the novel modified Tα1 developed in this study has the potential to be used for treating breast cancer.

Topics: Animals; Antineoplastic Agents; Apoptosis; Breast Neoplasms; Cell Adhesion; Cell Line, Tumor; Cell Proliferation; Female; Heterografts; Humans; Lymphocytes; MCF-7 Cells; Mice, Inbred BALB C; Mice, Nude; Molecular Conformation; Molecular Targeted Therapy; Oligopeptides; Spleen; Thymalfasin; Thymosin

Biomarkers predictive of pathological complete response (pCR) to neoadjuvant chemotherapy (NACT) of breast cancer are urgently needed.. Using a training/validation approach for detection of predictive biomarkers in HER2-negative breast cancer, pre-therapeutic core biopsies from four independent cohorts were investigated: Gene array data were analysed in fresh frozen samples of two cohorts (n=86 and n=55). Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed in formalin-fixed, paraffin-embedded (FFPE) samples from two neoadjuvant phase III trials (GeparTrio, n=212, and GeparQuattro, n=383).. A strong predictive capacity of thymosin beta 15 (TMSB15A) gene expression was evident in both fresh frozen cohorts (P<0.0001; P<0.0042). In the GeparTrio FFPE training cohort, a significant linear correlation between TMSB15A expression and pCR was apparent in triple-negative breast cancer (TNBC) (n=61, P=0.040). A cutoff point was then defined that divided TNBC into a low and a high expression group (pCR rate 16.0% vs 47.2%). Both linear correlation of TMSB15A mRNA levels (P=0.017) and the pre-defined cutoff point were validated in 134 TNBC from GeparQuattro (pCR rate 36.8% vs 17.0%, P=0.020). No significant predictive capacity was observed in luminal carcinomas from GeparTrio and GeparQuattro.. In TNBC, TMSB15A gene expression analysis might help to select patients with a high chance for pCR after NACT.

Topics: Breast Neoplasms; Cell Line, Tumor; Clinical Trials, Phase III as Topic; Estrogen Receptor alpha; Female; Gene Expression Profiling; Humans; Logistic Models; Receptor, ErbB-2; Receptors, Progesterone; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Thymosin

Intratumoral hypoxia has been correlated with distant metastatic potential. Two hypoxia inducible factors (HIFs), HIF-1α and HIF-2α, are induced by hypoxia, and high expression of these proteins has been correlated to angiogenesis and distant metastasis. Thymosin β4 (Tβ4) is frequently highly expressed in cancer, and this overexpression correlates with malignant progression. The objective of this study was to investigate the clinical correlation of HIF-α with Tβ4 and the intracellular functional roles of Tβ4 on HIF-α activation. We examined HIF-1α, HIF-2α and Tβ4 expressions in clinical human breast carcinoma (n=70) by immunohistochemistry. We show that high expression of HIF-1α and HIF-2α strongly correlates with Tβ4 expression (P≤0.0001) and overexpression of Tβ4 correlates significantly with patients with lymph node metastasis (P<0.05) of human breast cancer. Additionally, we demonstrate that hypoxia up-regulates intracellular Tβ4 protein, which then affects HIF-α activity, which is the key in regulating VEGF expression. We confirmed that hypoxia-induced intracellular Tβ4 and HIF-α activities were reduced by interference of Tβ4 expression using Tβ4 shRNA lentivirus. Vascular epidermal growth factor (VEGF)-A, a well-recognized lymphangiogenic cytokine, was also down-regulated, but VEGF-C and VEGF-D expressions were not affected. These findings suggest that the overexpression of Tβ4 is strongly associated with HIF-1α and HIF-2α expression and is also clinicopathologically involved with lymph node metastatic potential of breast cancer through the modulation of HIF-αactivation and induction of VEGF-A. Ultimately, these results highlight Tβ4 as a potentially therapeutic target in malignant cancers.

Topics: Basic Helix-Loop-Helix Transcription Factors; Biomarkers, Tumor; Blotting, Western; Breast Neoplasms; Cell Hypoxia; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Immunohistochemistry; Lymphatic Metastasis; Middle Aged; Reverse Transcriptase Polymerase Chain Reaction; Thymosin

Aromatase inhibitors can cause joint symptoms. The purpose of this pilot study was to evaluate the feasibility of immunologic therapies for this kind of joint symptoms.. A total of 16 postmenopausal women with stage I-III breast cancer with joint symptoms related to Aromatase inhibitors were enrolled. They received immunologic therapies of thymosin α1 1.6 mg, twice a week for 4 weeks. Outcome measures included the Brief Pain Inventory-Short Form, Western Ontario and McMaster Universities Osteoarthritis index, and the Functional Assessment of Cancer Therapy-General quality of life measure. Interferon-gamma and interleukin-4 were determined to evaluate immunomodulatory activity. Paired Samples Test and linear regression analysis were used to statistics the outcome measures.. From baseline to the end of treatment, patients reported improvement in the mean Brief Pain Inventory-Short Form worst pain scores (5.7-3.4, P < 0.001), pain severity (3.9-2.9, P = 0.01), and pain-related functional interference (4.2-1.8, P < 0.001), as well as the Western Ontario and McMaster Universities Osteoarthritis function subscale and Functional Assessment of Cancer Therapy-General physical well-being (P < 0.001 and P < 0.001, respectively). No adverse events were reported. The mean serum concentrations for secretion of interferon-gamma were significantly lower (P < 0.001); serum concentrations of interleukin 4 were higher (P = 0.02).. Immunologic therapies could play a role in reducing Aromatase inhibitor- related joint symptoms in breast cancer survivors and affecting the immune system in powerful ways. The improvements of immune system were associated with aromatase inhibitor-related joint symptoms.

Topics: Aromatase Inhibitors; Arthralgia; Breast Neoplasms; Dose-Response Relationship, Drug; Drug Administration Schedule; Feasibility Studies; Female; Follow-Up Studies; Humans; Immunotherapy; Pain Measurement; Patient Satisfaction; Pilot Projects; Severity of Illness Index; Survivors; Thymosin; Treatment Outcome

Beta-thymosins are polypeptides involved in the regulation of actin polymerization and thymosin beta10 and beta4 have been implicated in sequestration of monomeric (G-) actin. Additionally, experimental overexpression of thymosin beta10 has been found to result in increases in F-actin bundles as well as in cell motility and spreading. We have studied the distribution of endogenously expressed thymosin beta10 in cultured human breast cancer cell lines. Both unperturbed monolayer cultures and wound-healing models were examined using double-staining for thymosin beta10 and polymerized (F-) actin. Our findings show that thymosin beta10 is expressed in all three-cancer cell lines (SK-BR-3, MCF-7 and MDA-MB-231) studied. No or little staining was detected in confluent cells, whereas strong staining occurred in semiconfluent cells and in cells populating monolayer wounds. Importantly, the distribution of staining for thymosin beta10 was inverse of staining for F-actin. These data support a physiological role for thymosin beta10 in sequestration of G-actin as well as in cancer cell motility.

Topics: Actins; Breast Neoplasms; Cell Line, Tumor; Cell Movement; Cell Shape; Cytoskeleton; Female; Humans; Thymosin; Wound Healing

Previous studies have shown that the G-actin sequestering polypeptide thymosin beta4 frequently is overexpressed in cancers and that such overexpression correlates to malignant progression. However, the localization of thymosin beta4 in human cancers has not been determined. We now demonstrate that there is a considerable heterogeneity in the cellular distribution of thymosin beta4 in breast cancer. In most tumors examined, cancer cells showed low or intermediate reactivity for thymosin beta4, whereas leukocytes and macrophages showed intense reactivity. In addition, endothelial cells showed variable reactivity to thymosin beta4, whereas myofibroblasts were negative. There was no correlation between the intensity of tumor cell staining and histological grade, whereas there was a tendency toward a correlation between endothelial cell staining and grade. These results demonstrate that multiple cell types within the tumor microenvironment produce thymosin beta4 and that such expression varies from tumor to tumor. Such heterogeneity of expression should be taken into account when the role of thymosin beta4 in tumor biology is assessed.

Topics: Breast Neoplasms; Endothelial Cells; Female; Fluorescent Antibody Technique; Humans; Leukocytes; Macrophages; Stromal Cells; Thymosin

Overexpression of thymosin beta-4 has been linked to malignant progression but the localization of this polypeptide within tumors is incompletely known. We therefore examined breast cancers for thymosin beta-4 using immunofluorescence. Reactive cells were identified with monoclonal cell marker antibodies. A very heterogeneous staining pattern for thymosin beta-4 was observed. Thus, while leukocytes and macrophages showed intense reactivity for this polypeptide, cancer cells, and endothelial cells showed a much more variable reactivity. A similar heterogeneous staining was observed also in colorectal carcinomas. The degree of staining of breast cancer cells for thymosin beta-4 correlated neither to histological grade nor to endothelial cell staining. However, there was a tendency toward correlation (P = 0.07) between staining of endothelial cells and histological grade. Treatment of cultured breast cancer cells (SK-BR-3) with 1-4 microg thymosin beta-4/mL significantly increased cell numbers, as determined by MTT-assays. These data reveal an unexpected cellular heterogeneity of thymosin beta-4 expression in breast and colonic carcinomas and suggest that local release of this polypeptide in the tumor microenvironment may modulate tumor behavior.

Topics: Breast Neoplasms; Cell Nucleus; Colorectal Neoplasms; Cytoplasm; Female; Fluorescent Antibody Technique; Humans; Keratins; Leukocytes; Thymosin

Clinical trails showed that thymosin alpha1 offers protection from toxicities (nausea, vomiting, fatigue) of chemotherapy. This study was designed to investigate the protection of thymosin alpha1 to nervous system.. Twenty-two patients with advanced lung cancer, or advanced breast cancer were treated with vinorelbine (25 mg/m(2), d(1), d(8)) combined with cisplatin (80 mg/m(2), d(1)), or gemcitabine (1.25 g/m(2), d(1), d(8)) combined with cisplatin (80 mg/m(2), d(1)),or paclitaxel (80 mg/m(2), d(1), d(8), d(15)) combined with carboplatin (AUC=6 d(1)),or paclitaxel (80 mg/m(2), d(1), d(8), d(15)) combined with epirubicin (80 mg/m(2), d(1)). They all experienced grade 2 to 4 of neurotoxicities according to common toxicity criteria of National Cancer Institute after chemotherapy. The same chemotherapy regimens were combined with thymosin alpha1 (1.6 mg/d for 4 days before chemotherapy, and 1.6 mg twice weekly for 1-3 weeks after chemotherapy began) in the next cycle. Clinical neurologic evaluation was performed at baseline every week.. In 10 patients (45.4%), neurotoxicities reduced from grade 2-4 before chemotherapy to less than grade 2 after chemotherapy.. Thymosin alpha1 may prevent patients from chemotherapy-induced neurotoxicities.

Topics: Adult; Aged; Antineoplastic Combined Chemotherapy Protocols; Breast Neoplasms; Constipation; Female; Humans; Hypesthesia; Hypokinesia; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Neuroprotective Agents; Thymalfasin; Thymosin

A method has been developed for high sequence coverage analysis of proteins isolated from breast cancer cell lines. Intact proteins are isolated using multidimensional liquid-phase separations that permit the collection of individual protein fractions. Protein digests are then analyzed by both matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) peptide mass fingerprinting and by capillary electrophoresis-electrospray ionization (CE-ESI)-TOF MS peptide mapping. These methods can be readily interfaced to the relatively clean proteins resulting from liquid-phase fractionation of cell lysates with little sample preparation. Using combined sequence information provided by both mapping methods, 100% sequence coverage is often obtained for smaller proteins, while for larger proteins up to 75 kDa, over 90% coverage can be obtained. Furthermore, an accurate intact protein MW value (within 150 ppm) can be obtained from ESI-TOF MS. The intact MW together with high coverage sequence information provides accurate identification. More notably the high sequence coverage of CE-ESI-TOF MS together with the MS/MS information provided by the ion trap/reTOF MS elucidates posttranslational modifications, sequence changes, truncations, and isoforms that may otherwise go undetected when standard MALDI-MS peptide fingerprinting is used. This capability is critical in the analysis of human cancer cells where large numbers of expressed proteins are modified, and these modifications may play an important role in the cancer process.

Topics: Amino Acid Sequence; Breast Neoplasms; Cell Line, Tumor; Chromatography, High Pressure Liquid; Electrophoresis, Capillary; Electrophoresis, Gel, Two-Dimensional; Humans; Isoelectric Focusing; Lamins; Molecular Sequence Data; Molecular Weight; Protein Isoforms; Protein Processing, Post-Translational; Proteins; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Thymosin; Time Factors; Trypsin

Prothymosin alpha (PTalpha) is a small highly acidic protein found in the nuclei of virtually all mammalian tissues. Its high conservation in mammals and wide tissue distribution suggest an essential biological role. While the exact mechanism of action of PTalpha remains elusive, the one constant has been its relationship with the proliferative state of the cell and its requirement for cellular growth and survival. Recently PTalpha was found to promote transcriptional activity by sequestering the anticoactivator, REA from the Estrogen Receptor (ER) complex. We now report that Estradiol (E2) upregulates PTalpha mRNA and protein expression. Further studies indicate that ERalpha regulates PTalpha gene transcriptional activity. We have also delimited the region of PTalpha gene promoter involved in ERalpha-mediated transcriptional regulation and identified a novel ERalpha-binding element. Increased intracellular PTalpha expression in the presence of estrogens is accompanied by increased nuclear/decreased cytoplasmic localization. Increased nuclear expression of PTalpha is correlated with increased proliferation as measured by expression of Ki67 nuclear antigen. Conversely, inhibition of nuclear PTalpha expression in breast cancer cells using antisense methodology resulted in the inhibition of E2-induced breast cancer cell proliferation. Overall these studies underscore the importance of PTalpha in estrogen-induced breast cell proliferation.

Topics: Blotting, Northern; Blotting, Western; Breast Neoplasms; Cell Division; Chloramphenicol O-Acetyltransferase; DNA Primers; Electrophoretic Mobility Shift Assay; Estradiol; Estrogen Receptor alpha; Gene Deletion; Gene Expression Regulation, Neoplastic; Humans; Mutagenesis, Site-Directed; Polymerase Chain Reaction; Prohibitins; Promoter Regions, Genetic; Protein Precursors; Receptors, Estrogen; Retroviridae; RNA, Messenger; Thymosin; Transcription, Genetic; Tumor Cells, Cultured; Up-Regulation

To identify genes that are involved in breast cancer, suppression subtractive hybridization (SSH) was utilized to construct a breast cancer subtracted library. Differential screening of the library isolated 28 genes which by Northern analysis were highly expressed in the breast cancer cell line MDA-MB-231 compared to the normal breast cell line MCF12A. Sequence analysis revealed that 15 clones coded for previously described genes such as SNAP43, Cyr61, Thymosin beta4, tra1, elongation factor 1alpha, BSF-2/IL6, BiP, and GDP/GTP exchange protein. The remaining 13 clones did not match sequences in GenBank/EMBL database, indicating that they may be novel genes. SNAP43, a subunit of the TBP-TAF complex, was expressed 20-fold higher in MDA-MB-231 compared to MCF12A and several breast cancer cell lines, implying that SNAP43 may be involved in tumorigenesis of a specific subset of breast cancers. Amplification of SNAP43 was not found by Southern analysis. However, genetic alterations of MDA-MB-231 included a deletion of chromosome 14 with a reciprocal translocation t(6;14) and two additional translocations [t(12;14) and t(14;15)] as determined by fluorescent in situ hybridization (FISH) with YAC 823G8 located at chromosome 14q23 which contained SNAP43. Because of the numerous alterations observed by FISH in MDA-MB-231, we further explored the genetic abnormalities in this breast cancer cell line using multiplex FISH (M-FISH) and comparative genomic hybridization (CGH). These cells were replete with numerous complex structural rearrangements and had DNA copy-number imbalances involving multiple chromosomes including gains on chromosomes 2p, 2q31-q32, 3p14-pter, 5q, 6p, 7q36-qter, 11, 14q21-q24, 17p11.2-pter, 17q21-qter, 19, 20, Xp11-q13 and losses on chromosomes 4pter-q32, 8p, 9p21-p24, 10q26-qter, 16p13-pter, 18q12-qter, 22, Xp11.3-p22.1, Xq13-qter. In summary, SSH revealed a number of genes that were either novel or previously not associated with breast cancer. In addition, we found that breast cancer cells abounded with abnormalities as observed by M-FISH and CGH. Together, these results may facilitate defining the genetic alterations associated with breast cancer progression.

Topics: Breast Neoplasms; Chromosome Aberrations; Chromosomes, Human, Pair 4; Gene Expression Regulation, Neoplastic; Humans; In Situ Hybridization, Fluorescence; Nucleic Acid Hybridization; Thymosin; Transcription Factors; Translocation, Genetic; Tumor Cells, Cultured

Elevated expression of the beta-thymosin isotypes T beta(4), T beta(10), and T(15) appears to be involved in the manifestation of a malignant phenotype of human tumor cells, including those of mammary carcinomas. This has evoked an interest in these peptides as diagnostic/prognostic tumor markers. If increased levels of beta-thymosins correspond to tumor malignancy, the question arises whether tumor growth inhibition induced by chemotherapeutic drugs would reduce their expression.. Two human breast cancer cell lines, the estrogen receptor(ER)-positive MCF-7 and the ER-negative MDA-MB231, were thus analyzed for the amount of beta-thymosin mRNAs by RNase protection assay and for the respective peptide levels by HPLC following different hormonal and drug treatments.. Both cell lines, growing in medium with 10% FCS, contain T beta(4) (400-500 fg/cell) and Tbeta(10) (about 100 fg/cell), but no T beta(15). Incubating MCF-7 cells with tamoxifen (1 microM) for 5 days resulted in about 80% growth inhibition and in reduction of intracellular T beta(4) and T beta(10) concentrations by about 40%. Levels of T beta(4) and T beta(10)-mRNA were reduced by about 60%. In contrast, cisplatin (2 microM) changed neither the peptide concentrations nor the mRNA levels of beta-thymosins, in spite of marked growth inhibition. In addition, no changes in beta-thymosin expression were observed in MDA-MB231 cells treated with either drug. MCF-7 cells maintained in estrogen-poor medium (10% horse serum) or stimulated to grow with estradiol (1 nM) had Tbeta(4) and T beta(10) concentrations reduced by about 30%, but changes in T beta(4)- and T beta(10)-mRNA levels did not correspond to those of the peptide.. Expression of T beta(4) and T beta(10) mRNAs and their peptides is differentially regulated and does not correlate with growth. Instead, reduced beta-thymosin expression may be linked to more intensive TRITC-phalloidin staining of F-actin lining the membrane at sites of intimate cell-cell contacts, while increased beta-thymosin levels appear in cells with more extensive substrate adhesion. This suggests that beta-thymosins play a role in cell surface dynamics.

Topics: Actins; Antineoplastic Agents; Breast Neoplasms; Cytoskeleton; Female; Humans; Receptors, Estrogen; RNA, Messenger; Thymosin; Tumor Cells, Cultured

Prothymosin alpha (PTalpha), a protein associated with cell proliferation and chromatin remodeling, and found to selectively enhance ER transcriptional activity by interacting with a repressor of ER activity, is shown to be a primary response gene to estrogen. Prothymosin alpha mRNA was rapidly increased by estrogen, followed by a 6-fold increase in prothymosin alpha protein content in ER-containing breast cancer cells. Analysis of the prothymosin alpha promoter and 5'-flanking region, and electrophoretic gel mobility shift studies showed the strong inducibility by the estradiol-ER complex to be mediated by two consensus half-palindromic estrogen response elements at -750 and -1051, which directly bind the ER. Estrogenic stimulation of prothymosin alpha required a DNA binding form of ER with a functional activation function-2 domain. The prothymosin alpha 5'-regulatory region also contains multiple Sp1 sites. Although addition of Sp1 did not further enhance estradiol-ER stimulated prothymosin alpha transcriptional activity in breast cancer cells, transfection and response element mutagenesis studies using Drosophila cells, which are deficient in Sp1, revealed that Sp1 and the estradiol occupied-ER can each activate the prothymosin alpha gene independently of the other and act in an additive manner. These observations, documenting robust prothymosin alpha up-regulation by the estradiol-ER complex via widely spaced half-palindromic estrogen response element motifs, are reminiscent of those shown previously for the ovalbumin gene and suggest that the use of multiple half response elements may be a more common mode for regulation of gene expression by the ER than previously appreciated. In addition, these observations suggest interrelationships between cell proliferation and gene transcriptional activities and indicate a positive mechanism by which PTalpha, which increases ER transcriptional effectiveness, is itself up-regulated by the estrogen-ER complex.

Topics: Amino Acid Motifs; Breast Neoplasms; Estradiol; Estrogens; Gene Expression Regulation, Neoplastic; Humans; Mutagenesis, Site-Directed; Promoter Regions, Genetic; Protein Precursors; Receptors, Estrogen; Response Elements; RNA, Messenger; Sp1 Transcription Factor; Thymosin; Tumor Cells, Cultured; Up-Regulation

In a previous report we suggested that the estimation of prothymosin alpha (PTA) levels in primary breast tumours might be used to identify breast cancer patients at high risk for distant metastasis (Dominguez F et al (1993) Eur J Cancer 29A: 893-897). Here the role of tumour PTA levels as predictor was investigated with respect to both disease-free survival (DFS) and survival. Tumours were obtained from a series of 210 consecutive female patients with ductal carcinoma who underwent surgery at the Hospital Xeral de Galicia (Santiago de Compostela, Spain). Characteristics including PTA tumour levels, number of positive axillary nodes, patient's age at surgery and tumour histological grade were significantly associated with DFS and survival, as determined by univariate analysis. Patients with tumours with low or moderate PTA levels demonstrated a statistically decreased rate of tumour recurrence and a statistically significant increased overall survival in comparison with those whose tumours had high PTA levels. Patient's relative risk of dying was 2.1 times greater for tumours with high PTA levels than for those tumours with low or moderate PTA levels. In conclusion, these data support the hypothesis that tumour high PTA levels is associated with a worse outcome.

Topics: Adult; Biomarkers, Tumor; Breast Neoplasms; Disease-Free Survival; Humans; Middle Aged; Outcome Assessment, Health Care; Prognosis; Protein Precursors; Thymosin

Peripheral blood mononuclear cells (PBMC) from cancer patients were cultured in vitro with irradiated autologous tumor cells isolated from malignant effusions (mixed lymphocyte tumor cultures, MLTC) and low-dose (50 IU/ml) recombinant interleukin-2 (IL-2). The combination of IL-2 and prothymosin alpha (ProTalpha) resulted in a greater PBMC-induced response to the autologous tumor than that brought about by IL-2 alone. In particular, ProTalpha specifically enhanced the CD4+ T-cell-mediated proliferation against the autologous tumor. CD4+ T cells seemed to recognize tumor antigens presented by HLA-DR molecules expressed on the autologous monocytes, since preincubation of the latter with an anti-HLA-DR monoclonal antibody (mAb) abrogated the response. In addition, MLTC set up with IL-2 and ProTalpha also generated more MHC-class-I-restricted cytotoxic T lymphocytes (CTL) against the autologous tumor than did MLTC set up with IL-2 alone. The MLTC-induced CTL contained high levels of cytoplasmic perforin and their development was strictly dependent on the presence of both autologous CD4+ T cells and monocytes. In the absence of either population there was a strong impairment of both proliferative and cytotoxic responses which was not restored by the presence of ProTalpha. In contrast, when both cell populations were present, ProTalpha exerted optimal enhancement of CD4+ T cell proliferation, which was associated with potentiated CTL responses. Our data emphasize the role of ProTalpha for the enhancement of IL-2-induced CTL responses against autologous tumor cells. Such responses require collaborative interactions between CD4+, CD8+ T cells and monocytes as antigen-presenting cells. Our data are relevant for adoptive immunotherapeutic settings utilizing IL-2 and ProTalpha-induced autologous-tumor-specific CTL.

Topics: Aged; Animals; Breast Neoplasms; Carcinoma; Cattle; CD3 Complex; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Communication; Cell Division; Cell Separation; Cells, Cultured; Female; Humans; Interleukin-2; Lung Neoplasms; Male; Middle Aged; Monocytes; Ovarian Neoplasms; Phenotype; Protein Precursors; T-Lymphocytes, Cytotoxic; Thymosin; Thymus Gland; Time Factors; Tumor Cells, Cultured

The beta-thymosins comprise a family of structurally related, highly conserved acidic polypeptides, originally isolated from calf thymus. Recently, we have demonstrated the overexpression of thymosin beta-10 (TB10) in rat thyroid transformed cell lines and in human thyroid carcinoma tissues and cell lines. To verify whether TB10 overexpression is a general event in the process of carcinogenesis, we have analyzed TB10 mRNA levels in human colon carcinomas, germ cell tumors of different histological types, breast carcinomas, ovarian carcinomas, uterine carcinomas, colon and esophageal carcinoma cell lines. Overexpression of the TB10 gene was detected in all of the neoplastic tissues and cell lines compared to the respective normal tissues. Moreover, the mouse model of skin carcinogenesis induced by the combined action of chemical carcinogens and phorbol esters was used to identify the stage of TB10 gene induction. The expression was almost undetectable in normal keratinocytes, its induction occurred even at the papilloma stage, however a further increased expression was observed in the carcinoma derived cell lines. Finally, immunohistochemical analysis of some breast, colon and ovary carcinoma samples by using specific anti-TB10 antibodies revealed the presence of the TB10 protein in all of the neoplastic tissues, but not in the respective normal tissues. Therefore the TB10 detection may be considered a potential tool for the diagnosis of several human neoplasias.

Topics: Animals; Breast Neoplasms; Carcinoma; Colonic Neoplasms; Esophageal Neoplasms; Female; Gene Expression; Germinoma; Humans; Immunohistochemistry; Male; Mice; Neoplasms; Ovarian Neoplasms; RNA; Skin Neoplasms; Testicular Neoplasms; Thymosin; Tumor Cells, Cultured; Uterine Neoplasms

The myc proto-oncogenes are transcription factors that directly regulate the expression of other genes, by binding to the specific DNA sequence, CACGTG. Among the target genes for c-Myc regulation are ECA39, p53, ornithine decarboxylase (ODC), alpha-prothymosin and Cdc25A. In this study we examined the involvement of c-Myc target genes in human oncogenesis induced by c-myc or N-myc. In MCF-7 breast cancer cells, the induction of c-myc expression by estrogen was followed by the induction of all the Myc targets that we examined, indicating that those genes can serve as c-Myc targets in human oncogenesis. Moreover, in breast tumors exhibiting c-myc overexpression, several Myc targets were also overexpressed. A clear correlation between the expression of c-myc and its targets was also detected in Burkitt's lymphomas, which involve a specific translocation of c-myc gene, but not in other lymphoma cells. Yet, in cells derived from a neuronal origin the pattern of expression of Myc targets was more complex. In a neuroepithelioma cell line that overexpresses c-myc, only some targets were expressed. In addition in neuroblastomas, in which N-myc is amplified and overexpressed, only ODC was overexpressed in all cell lines, while all other target genes were expressed in only some of the cell lines. The more complex expression pattern found for the Myc targets in neuroblastomas suggests that genes that were identified originally as targets for c-Myc regulation may be regulated by N-Myc, but other cell specific factors are also needed for transcription of the target genes.

Topics: Breast Neoplasms; Burkitt Lymphoma; cdc25 Phosphatases; Cell Transformation, Neoplastic; Estradiol; Female; Gene Expression Regulation, Neoplastic; Humans; Neuroblastoma; Neuroectodermal Tumors, Primitive, Peripheral; Ornithine Decarboxylase; Protein Precursors; Protein Tyrosine Phosphatases; Proteins; Proto-Oncogene Proteins c-myc; Thymosin; Transaminases

An enzyme linked immunosorbent assay, specific for prothymosin alpha (ProT alpha) was developed using an antibody against the synthetic C-terminal peptide ProT alpha[101-109] and isolated bovine ProT alpha for the preparation of standard solutions and immunoplates. Due to the antibody used, the ELISA developed was capable of fully discriminating between ProT alpha, the naturally occuring and partially homologous peptide parathymosin alpha (ParaT alpha) and the peptide thymosin alpha1 (T alpha1), whose sequence is identical to the [1-28] sequence of ProT alpha, and its in vivo occurrence is under question. Moreover, due to its improved sensitivity, the ELISA was capable of directly determining ProT alpha concentration in human serum and tissue extracts, without any pretreatment of the samples. ProT alpha levels were directly measured in sera obtained from 48 apparently healthy individuals and 27 patients with diagnosed breast cancer and found to range from 0.67 to 2.34 microg/ml (mean value 1.27 +/- 0.49 microg/ml) and from 0.47 to 1.74 microg/ml (mean value 1.02 +/- 0.29 microg/ml), respectively. ProT alpha levels were also measured in four breast tumor and adjacent normal breast tissue extracts and found to be elevated in the tumor extracts.

Topics: Adolescent; Adult; Animals; Antibodies; Biomarkers; Breast; Breast Neoplasms; Enzyme-Linked Immunosorbent Assay; Epitopes; Female; Humans; Male; Middle Aged; Protein Precursors; Rabbits; Sensitivity and Specificity; Thymosin; Titrimetry

Synaptophysin and/or chromogranin A may be expressed in epithelial cells of normal and dysplastic mammary gland and in some cancer of the breast. This work indicates that some stromal cells form thin walled vascular channels and from their perivascular mesenchyma arise myoid-like cells, some with cross-like striations. These myoid-like cells have been stained with antibody to smooth muscle actin, rarely to desmin and sarcomeric actin as well as are strongly positive for synaptophysin and/or chromogranin A. From those cells arise cancer cells. Some cancer cells also expressed desmin, sarcomeric actin or smooth muscle actin. Because synaptophysin positive are neuromuscular endplates, it is a question whether the presence of synaptophysin in the progenitor of neoplastic cells of myogenic origin is due to the synaptic dysfunction in molecular mechanism of the epithelial-parenchyma and mesenchymal stroma interaction. The location of S100-protein positive dendritic cells distributed in regular pattern in suprabasal layer of the mammary gland indicate that these cells may arise from preadipocytes which take part in morphogenesis of the breast. All cases of the breast cancer demonstrated thymosin alpha 1, some of them also showed Hassall's-like bodies, mucin secretion and minute calcification. Whether the interaction of epithelial cells and S100-protein positive dendritic cells in tissue other than the thymus has similar dynamic activity to that of Hassall's bodies of the thymus remains to be determined. Both, epithelial cells of Hassall's bodies and epithelial cells of the investigated breast are of myogenic origin. The above consideration suggests that the cells forming vascular channels are with a differentiation defect as they are created in altered stromal mesenchyma and the presence of thymic growth factors might induce tumor growth.

Topics: Adjuvants, Immunologic; Biomarkers, Tumor; Breast Neoplasms; Carcinoma; Chromogranin A; Chromogranins; Disease Progression; Humans; Immunohistochemistry; Prognosis; S100 Proteins; Synaptophysin; Thymalfasin; Thymosin

Thymosin beta15 is a newly discovered 5300-Da protein that binds actin monomers and inhibits actin polymerization and might thus increase cellular motility. Thymosin beta15 is upregulated at both the mRNA and protein levels in prostate cell lines in a manner directly related to their capacity to metastasize. We hypothesize that because this protein is upregulated in cells with a propensity to metastasize, it might be a useful prognostic marker in breast cancer. Because this is a newly described protein, neither the subcellular localization of thymosin beta15 or its expression in breast cancer has been examined. We describe the use of an affinity-purified polyclonal antibody to show that within breast epithelium, thymosin beta15 is localized diffusely throughout the cytoplasm and that thymosin beta15 is upregulated in malignant (compared with benign) breast tissue. In contrast to the prostate model, thymosin beta15 is upregulated in nonmetastatic breast cancer and even ductal carcinoma in situ (compared with benign breast tissue), and, consequently, it might represent a potential early marker for breast malignancy. Additional studies are needed to evaluate the precise role and prognostic value of thymosin beta15 in breast cancer.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Breast; Breast Diseases; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Female; Humans; Immunoenzyme Techniques; Middle Aged; Prognosis; Thymosin; Up-Regulation

Immunoregulatory effects of thymic peptides on functions of polymorphonuclear leukocytes (PMNs) are poorly investigated. We studied the effects of prothymosin alpha 1 (Pro alpha 1) on PMNs from patients with colorectal tumors, breast tumors and melanoma (total n = 37) in comparison with healthy donors (n = 18), with respect to chemotaxis, cytotoxicity against HCT-116 colon tumor cells, oxidative response (chemiluminescence reaction) as well as expression of surface marker molecules. We found that Pro alpha 1 was equally effective in stimulating the chemotactic activity of PMNs from tumor patients and healthy donors (43% increase). PMNs from tumor patients, especially with breast tumor, showed a significant enhancement of cytotoxicity against the tumor target cells in comparison with healthy donors. With respect to the PMNs cytotoxicity, only about 50% of the colorectal tumor patients and healthy donors responded to Pro alpha 1 and FMLP. As to the oxidative response of PMNs, elevated levels were found only among colorectal tumor patients. Pro alpha 1 significantly increased the oxidative response in breast and colorectal tumor patients by 55% and 25%, respectively. Pro alpha 1 decreased the expression of CD16 on PMNs of healthy donors, but not that of CD11a, CD11b, CD11c, CD13, CD14, CD15 and CD32. Therefore, we suggest, that Pro alpha 1 may improve some PMN functions of tumor patients, associated with the proposed role in host-tumor interaction.

Topics: Aged; Aged, 80 and over; Breast Neoplasms; Chemotaxis, Leukocyte; Cytotoxicity, Immunologic; Female; Humans; Luminescent Measurements; Macrophage-1 Antigen; Male; Melanoma; Middle Aged; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Protein Precursors; Reactive Oxygen Species; Receptors, IgG; Thymosin

Paraffin sections from 30 human breast tissue specimens were stained with a specific antibody for thymosin beta-10, ATB10(38-43). The results showed that thymosin beta-10 was detected mainly in the malignant tissue, particularly in the cancerous cells, whereas the normal cell population around the lesions showed very weak staining. Also, the intensity of staining in the cancerous cells was proportionally increased with the increasing grade of the lesions.

Topics: Adult; Aged; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Enzyme-Linked Immunosorbent Assay; Female; Humans; Middle Aged; Neoplasm Proteins; Thymosin

Thymosin alpha 1 (T alpha 1) is an immune modulatory peptide which has been evaluated in a variety of clinical trials. Although no in vivo adverse effects, including enhancement of tumor growth, have been noted, in vitro studies suggesting a role for T alpha 1 in cell growth have been reported. The studies presented in this report evaluated both exogenously added T alpha 1 and endogenously expressed T alpha 1 as factors which could either promote growth of tumor cells or induce transformation. No effect of exogenous T alpha 1 on cell growth was found. NIH-3T3 cells transfected with cDNA for the precursor ProThymosin alpha (Pro T alpha) expressed elevated levels of authentic T alpha 1 but did not demonstrate either enhanced proliferation in liquid culture or transformation as defined by the loss of contact inhibition or anchorage independent growth in soft agar. Thus these studies argue against the hypothesis that T alpha 1 is either an intracellular or extracellular growth promoter.

Topics: 3T3 Cells; Adenocarcinoma; Animals; Breast Neoplasms; Cecal Neoplasms; Cell Division; Cell Transformation, Neoplastic; Colonic Neoplasms; Growth Substances; Ileal Neoplasms; Mice; Protein Precursors; Thymosin; Tumor Cells, Cultured

A radioimmunoassay specific for the C-terminus of human prothymosin alpha was developed using the synthetic peptide [Cys-Aca degrees]-human prothymosin alpha (90-109)-OH coupled to KLH as antigen and the analogue [Tyr-Aca degrees]-human prothymosin alpha (90-109)-OH labelled with 125I as tracer. The radioimmunoassay measured intact prothymosin alpha, in the range of 2-100 pmol and does not cross-react with the partly homologous polypeptide parathymosin alpha. A major epitope was located in the segment 95-107. A radioimmunoassay specific for the N-terminus of human parathymosin alpha, also measuring intact parathymosin alpha in the range of 1-20 pmol and not cross-reacting with prothymosin alpha, was developed using the synthetic peptide [Cys-Aca degrees]-human parathymosin alpha (1-30)-OH as antigen coupled to KLH and the analogue [Tyr-Aca degrees]-human parathymosin alpha (1-10)-OH labelled with 125I as tracer. A major epitope was located in the segment 1-10. These radioimmunoassays, together with a previously established radioimmunoassay for the N-terminus of prothymosin alpha, permitted the identification of the molecular forms of the cross-reactive materials in both normal and neoplastic breast tissue extracts as intact prothymosin alpha and parathymosin alpha. It was also possible to reveal significantly higher levels of both alpha-thymosins in breast cancer tissue compared to the nearby healthy tissue--the mean of 14 samples was over 14-fold higher--suggesting a role of both prothymosin alpha and parathymosin alpha in cell proliferation. The reported radioimmunoassays are expected to facilitate the search for prognostic and/or diagnostic applications of these polypeptides in human cancer.

Topics: Aged; Aged, 80 and over; Amino Acid Sequence; Animals; Breast Neoplasms; Chromatography, High Pressure Liquid; Cross Reactions; Epitopes; Female; Humans; Middle Aged; Molecular Sequence Data; Peptides; Protein Precursors; Rabbits; Radioimmunoassay; Thymalfasin; Thymosin

In 71 patients with classic invasive ductal carcinomas, levels of prothymosin alpha (PT alpha), as assayed by a radioimmunoassay that detects thymosin alpha 1 (the NH2-terminal fragment of PT alpha), were significantly greater in tumour samples than in normal breast tissue. PT alpha levels were correlated with (a) the number of positive axillary lymph nodes (rs = 0.5384, P < 0.01), and (b) the percentage of tumour cells in the S or G2/M phase as assessed by flow cytometry (rs = 0.5027, P < 0.01). Since the beginning of this study in 1989, 21 patients have presented distant metastases, all of whom were previously shown to have tumour PT alpha levels greater than 124 ng of thymosin alpha 1/mg protein. The present report indicates that PT alpha might be used to identify breast cancer patients at high risk for distant metastases.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Carcinoma, Intraductal, Noninfiltrating; Female; Humans; Lymphatic Metastasis; Middle Aged; Mitosis; Prognosis; Protein Precursors; Radioimmunoassay; Thymosin

Topics: Adjuvants, Immunologic; Antineoplastic Combined Chemotherapy Protocols; Breast Neoplasms; Cyclophosphamide; Dactinomycin; Doxorubicin; Female; Fluorouracil; Humans; Methotrexate; Neutrophils; Peptides; Phagocytosis; T-Lymphocytes; Thymosin; Thymus Extracts; Vincristine

Growth of normal and malignant cells is controlled by the interplay of several hormones and growth factors present in the body. The growth of the human breast cancer cell line MCF-7 is stimulated in vitro by estradiol which has been shown to induce the release of numerous polypeptide growth factors into the culture media. In a search to identify polypeptide growth factors for MCF-7 breast cancer cells we have detected the thymic hormone, thymosin alpha one (TA1) in culture supernatants and cytosol preparations of MCF-7 cells grown in TA1 free media. TA1 was identified by reverse phase-high performance liquid chromatography (RP-HPLC) followed by a specific radioimmunoassay for TA1 (TA1-RIA). Indirect immune fluorescence localized TA1 on, or within, the cytoplasm of MCF-7 cells grown in TA1 free media. The results of this data along with our previously published findings fulfills three of the four criteria needed to establish thymosin alpha-1 as an autocrine growth factor for MCF-7 cells in culture.

Topics: Breast Neoplasms; Cell Line; Chromatography, High Pressure Liquid; Female; Fluorescent Antibody Technique; Humans; Radioimmunoassay; Thymalfasin; Thymosin; Tumor Cells, Cultured

Thymosins beta 4 and beta 10 are 2 structurally related polypeptides originally defined in the rat immune system. To date, no truly unambiguous functions have been formally ascribed to these small (less than 4.9 kDa) acidic proteins. Previous research has demonstrated relationships between expression of these genes and cell growth/differentiation. These observations prompted the present study which has used cDNA and synthetic oligonucleotide probes in combination with high-performance liquid chromatography (HPLC) to examine the differential expression of these 2 genes in normal and neoplastic human tissues and in the developing human kidney. Low levels of beta 4 and beta 10 mRNA species prevailed in normal tissues; in contrast, these gene transcripts were notably more abundant in malignant renal tumors and in the normal human embryonic kidney. These findings show that the thymosin beta 4 and beta 10 genes are constitutively expressed at higher levels in embryonic/neoplastic as compared to normal/benign tissues and that thymosin in beta 10 in particular may be a new molecular marker for renal-cell carcinoma as well as other malignancies.

Topics: Aging; Base Sequence; Blotting, Northern; Breast Neoplasms; Chromatography, High Pressure Liquid; Cloning, Molecular; Female; Gene Expression; Humans; Infant; Kidney; Kidney Neoplasms; Middle Aged; Molecular Sequence Data; Neoplasm Proteins; Neoplasms; Nucleic Acid Hybridization; Oligonucleotide Probes; Ovarian Neoplasms; Reference Values; RNA, Messenger; Thymosin

Topics: Adult; B-Lymphocytes; Breast Neoplasms; Dose-Response Relationship, Immunologic; Humans; Kidney Neoplasms; Melanoma; Middle Aged; Neoplasms; Rosette Formation; T-Lymphocytes; Thymosin; Thymus Hormones

A Phase I clinical trial of thymosin administered in doses of 10 to 250 mg/M2 intramuscularly for seven days was undertaken in ten patients with disseminated malignancies and evidence of immunoincompetence. Toxicity was minimal; one patient experienced a mild urticarial rash which cleared spontaneously, two patients developed low grade fever and one patient experienced pain at the injection site. There was no evidence of systemic toxicity or parenchymal organ dysfunction. Thymosin administration was associated with an increase in the E-rosette forming capacity of the patient's lymphocytes and the development of new skin test reactivity to recall antigens in some of these patients. One objective tumor response was noted. These findings are preliminary but are encouraging for further utilization of thymosin as an immunostimulant in cancer patients with immunoincompetence.

Topics: Adenocarcinoma; Breast Neoplasms; Drug Eruptions; Dysgerminoma; Hodgkin Disease; Humans; Immune Adherence Reaction; Immunity, Cellular; Leiomyosarcoma; Lung Neoplasms; Lymphocytes; Melanoma; Neoplasm Metastasis; Neoplasms; Skin Tests; Thymosin; Thymus Hormones

The effect of thymosin on in vitro reactivity of peripheral lymphocytes to phytohemagglutinin, concanavalin A, and the formation of spontaneous E-rosettes in 54 patients with metastatic carcinomas has been studied. Thymosin increased lymphocyte responses to PHA and Con A in only 10 patients, with predominant effect seen with Con A. Twenty patients showed depressed baseline levels of E-rosettes which were increased to normal or subnormal levels after incubation with thymosin. No distinct correlation was noted between the clinical stage of the disease and the ability of lymphocytes to form E-rosettes. Although the exact mechanism by which the thymus exerts its influence on host resistance is not clearly defined, present evidence supports the concept that the thymic hormone, thymosin, may be an important addition in treatment of cancer patients by increasing cell-mediated immunity.

Topics: Adolescent; Adult; Aged; Breast Neoplasms; Carcinoma, Bronchogenic; Concanavalin A; Hodgkin Disease; Humans; Immune Adherence Reaction; In Vitro Techniques; Lectins; Middle Aged; Neoplasms; T-Lymphocytes; Thymosin; Thymus Extracts

In preparation for the use of bovine thymosin, a thymic hormone, as a specific T cell stimulator in immunodepressed patients, we studied its effect on E rosette formation of peripheral lymphocytes from patients with (1) advanced malignancies, (2) extensive burns, and (3) septicemia. E rosette formation in vitro with and without thymosin was evaluated in 52 patients with carcinoma of the breast (25) or lung (27) in relation to adjuvant therapy and/or surgery. The depression of E rosettes in cancer patients was most striking when adjuvant therapy, irradiation, and/or chemotherapy were used; in 20 patients this was elevated by incubation with thymosin. There was a delay in recovery of depressed E rosette levels after radical mastectomies in four patients, recovery being accelerated by thymosin. In ten burn patients (40 to 80 percent of body surface area, second and third degree burns), the depression in E rosette levels occurred in the first week and was most marked in 3 to 4 weeks. In eight patients this was elevated by thymosin in vitro. In four septic patients, all undergoing operation, serial studies suggested that recovery from sepsis was accompanied by spontaneous rise in E rosette levels. This process was accelerated by thymosin in vitro. This study as well as previous experiments with animals suggest that thymosin may influence depressed host resistance favorably by increasing T-cell-mediated immunity.

Topics: Adolescent; Adult; Aged; Breast Neoplasms; Burns; Child; Child, Preschool; Female; Humans; Immune Adherence Reaction; Immunologic Deficiency Syndromes; In Vitro Techniques; Lung Neoplasms; Male; Middle Aged; Sepsis; T-Lymphocytes; Thymosin; Thymus Extracts