Immunolabelling of the inhibin/activin-ßC subunit in normal and malignant human uterine cervical tissue and cervical cancer cell lines.

Inhibins are dimeric glycoproteins, composed of an α-subunit and one of two possible ß-subunits (ßA or ßB), with substantial roles in human reproduction and in endocrine-responsive tumours. Recently a novel ß subunit named ßC was described, although it is still unclear if normal or cancerous cervical epithelial cells as well as cervical cancer cell lines can synthesise the inhibin-ßC subunit. Four normal cervical tissue samples together with specimens of well-differentiated squamous cervical cancer and adenocarcinoma of the cervix were immunohistochemically analyzed. Additionally, two cervical carcinoma cell lines (HeLa and CaSKi) were analyzed by immunofluorescence for the expression of this novel subunit. We demonstrated for the first time an immunolabelling of the inhibin-ßC subunit in normal and malignant cervical tissue, as well as cervical cancer cells. Although the physiological role is still unclear in cervical tissue, the inhibin-ßC subunit might play important roles in carcinogenesis. Moreover, the synthesis of this subunit in cervical carcinoma cell lines of squamous and epithelial origins allows the use of these cell lines in elucidating its functions in cervical pathogenesis.

Evaluation of two commercialised in situ hybridisation assays for detecting HPV-DNA in formalin-fixed, paraffin-embedded tissue.

INTRODUCTION: The role of human papilloma virus (HPV) in the pathogenesis of anogenital dysplasia is now conclusive. However, HPV detection in formalin-fixed and paraffin-embedded tissues remains controversial. Therefore, the aim of this study was to evaluate morphological changes directly in tissue specimens using a HPV-DNA detection system involving HPV in situ hybridisation. MATERIALS AND METHODS: Samples from patients with cervical carcinoma were analysed using the GenPoint HPV DNA Probe Cocktail (Dako, Glostrup, Denmark) and the ZytoFast HPV Screening CISH-Kit (Zytomed, Berlin, Germany). Three cervical carcinoma cell lines with a well-defined HPV copy number per cell (SiHa, HeLa, and CaSki) served as positive controls for sensitivity testing, while two HPV-negative cell lines (AC-1M32, MCF-7) and brain tissue samples served as negative controls. Moreover, to assess the validity of the in situ hybridisation, the expression of HPV-16 DNA in cell lines was demonstrated by HPV-16 E6-specific PCR. RESULTS: Both HPV-screening assays revealed strong signals of episomal and integrated HPV-DNA at a HPV copy number of more than 50 copies/cell. All cervical carcinoma samples were positive in the Dako assay, which identifies 13 high-risk HPV genotypes, whereas HPV-DNA could be detected in 9/10 cervical carcinoma samples using the Zytofast assay, identifying HPV 16, 18, 31, 33, and 35. CONCLUSION: HPV in situ hybridisation is a convenient and powerful tool for detecting HPV-DNA in formalin-fixed and paraffin-embedded tissue samples. Therefore, this technique is suitable for analysis of a potential HPV infection using archival pathological slides.

Inhibin-betaA and -betaB subunits in normal and malignant glandular epithelium of uterine cervix and HeLa cervical cancer cell line.

INTRODUCTION: Inhibins, dimeric peptide hormones composed of an alpha-subunit and one of two possible beta-subunits (betaA or betaB), exhibit substantial roles in human reproduction and in endocrine-responsive tumors. However, it is still unclear if normal and cancerous cervical glandular epithelial cells as well as cervical cancer cell lines of glandular origin express the inhibin-betaA and -betaB subunits. MATERIALS AND METHODS: Normal cervical tissue samples and a total of 10 specimens of well-differentiated adenocarcinomas of the human cervix were analyzed for inhibin-betaA and -betaB subunit expression by immunohistochemical analysis. Additionally, the cervical carcinoma cell line HeLa was analyzed by immunofluorescence and RT-PCR analysis for the expression of inhibin subunits. RESULTS: Immunolabeling of normal and malignant glandular epithelium of human cervical tissue revealed a positive staining reaction for the inhibin-betaA and -betaB subunits. Additionally, the cancer cell line HeLa synthesized both inhibin subunits. When compared to the normal cervical glandular epithelium, the expression of the inhibin beta subunits became significantly reduced in cervical adenocarcinoma tissues. DISCUSSION: In conclusion, we demonstrated a strong, though differential expression pattern of inhibin-betaA and -betaB subunits in normal and malignant glandular epithelial cells of the human uterine cervix. Although the physiological role of inhibins is still quite unclear in cervical tissue, the expression of inhibin-beta-subunits might play an important role in cervical cancer carcinogenesis, since they are significantly down-regulated during pathogenesis in cervical adenocarcinomas.

Inhibin/activin-betaC subunit does not represent a prognostic parameter in human endometrial cancer.

INTRODUCTION: Inhibins, dimeric peptide hormones composed of an α subunit and one of two possible ß subunits (betaA or betaB), exhibit substantial roles in human reproduction and in endocrine-responsive tumors. Recently, two novel inhibin-beta subunits, defined as betaC and betaE, have been identified in humans. However, the prognostic significance and clinical implications of the novel inhibin-betaC subunit in endometrial cancers is still quite unclear. MATERIALS AND METHODS: A series of 296 uterine endometrial carcinomas were immunohistochemically analyzed with specific antibody against the inhibin-betaC subunit. The staining reactions were correlated with several clinicopathological characteristics and the clinical outcome. RESULTS: Endometrial cancer tissue demonstrated an immunolabelling against the inhibin-betaC subunit. The inhibin-betaC expression in endometrial carcinoma samples revealed a significant association with hemangiosis. However, the expression of this inhibin subunit did not affect patients' progression-free, cause-specific and overall survival. CONCLUSION: Overall, inhibin-betaC subunit was demonstrated in endometrial cancer tissue. This novel betaC subunit demonstrated a significant association with hemangiosis although without any impact on the patients' survival. Moreover, the inhibin-betaC subunits did not constitute an independent prognostic parameter in endometrial cancer patients. Therefore, the isolated analysis of this subunit might be of minor prognostic value in identifying high-risk patients.

Evidence of inhibin/activin subunit betaC and betaE synthesis in normal human endometrial tissue.

BACKGROUND: Inhibins are important regulators of the female reproductive system. Recently, two new inhibin subunits betaC and betaE have been described, although it is unclear if they are synthesized in normal human endometrium. METHODS: Samples of human endometrium were obtained from 82 premenopausal, non-pregnant patients undergoing gynecological surgery for benign diseases. Endometrium samples were classified according to anamnestic and histological dating into proliferative (day 1-14, n = 46), early secretory (day 15-22, n = 18) and late secretory phase (day 23-28, n = 18). Immunohistochemical analyses were performed with specific antibodies against inhibin alpha (n = 81) as well as inhibin betaA (n = 82), betaB (n = 82), betaC (n = 74) and betaE (n = 76) subunits. RT-PCR was performed for all inhibin subunits. Correlation was assessed with the Spearman factor to assess the relationship of inhibin-subunits expression within the different endometrial samples. RESULTS: The novel inhibin betaC and betaE subunits were found in normal human endometrium by immunohistochemical and molecular techniques. Inhibin alpha, betaA, betaB and betaE subunits showed a circadian expression pattern, being more abundant during the late secretory phase than during the proliferative phase. Additionally, a significant correlation between inhibin alpha and all inhibin beta subunits was observed. CONCLUSIONS: The differential expression pattern of the betaC- and betaE-subunits in normal human endometrial tissue suggests that they function in endometrial maturation and blastocyst implantation. However, the precise role of these novel inhibin/activin subunits in human endometrium is unclear and warrants further investigation.

Immunolabeling of the inhibin-ßA and -ßB subunit in normal and malignant human cervical tissue and cervical cancer cell lines.

OBJECTIVES: Inhibins, dimeric peptide hormones composed of an α-subunit and 1 of 2 possible ß subunits (ßA or ßB), exhibit substantial roles in human reproduction and in endocrine-responsive tumors. However, it is still unclear whether normal and cancerous cervical tissues as well as cervical cancer cell lines express the inhibin-ßA and -ßB subunits. MATERIALS AND METHODS: Normal human uterine cervical tissue was obtained from 4 premenopausal nonpregnant patients. In addition, a total of 32 specimens of cervical intraepithelial neoplasia (CIN) of different stages were obtained (CIN 1 = 10, CIN 2 = 9, and CIN 3 = 13). Moreover, 30 squamous cervical cancer samples of well-differentiated (grade 1; n = 10), moderate differentiated (grade 2; n = 10), and poorly differentiated (grade 3; n = 10) grading were analyzed. RESULTS: An immunohistochemical staining reaction for inhibin-ßA and -ßB subunits could be observed in normal and malignant cervical tissue as well as in cervical cancer cell lines. Regarding inhibin-ßA significant differences were observed between normal tissue and CIN 1 and CIN 3. Moreover, the immunohistochemical staining reaction for inhibin-ßA was significantly higher in CIN 3 compared with that in cervical carcinoma grades 1 and 2. The inhibin-ßB expression was higher in CIN and cervical cancer compared with that in normal cervical tissue. Inhibin-ßB was significantly higher in CIN 2 and CIN 3 compared with cancer tissues of histological grade 1. In addition, a significant increase of the staining intensity was observed between cervical cancer grades 1 and 2 as well as grade 3. CONCLUSIONS: Both inhibin-ß subunits demonstrated a differential expression in CIN and squamous cancer, suggesting important roles in cervical carcinogenesis. Inhibin-ßA might be important during progression of CIN, whereas the inhibin-ßB subunit could exert a substantial function during differentiation of cervical carcinomas. Moreover, the synthesis of this subunit in cervical carcinoma cell lines also allows the use of this cell line to elucidates their functions in cervical cancer pathogenesis.

Prognostic significance of estrogen receptor alpha and beta expression in human serous carcinomas of the ovary.

BACKGROUND: Ovarian cancer is a gynecologic cancer with a high mortality rate. This shows the need of effective screening methods and reliable cancer markers for ovarian cancer staging and appropriate effective cancer treatment. METHODS: Expression of estrogen receptor alpha and beta was studied by immunhistochemical analysis of 100 ovarian cancer tissue samples. RESULTS: In this study on 100 serous ovarian cancer tissue samples from patients with defined clinicopathologic features, a significant association of ER-alpha expression with ovarian cancer grading (p = 0.004), progression-free survival (p = 0.007), and cause-specific survival (p = 0.001) was demonstrated. Expression of ER-beta was found to be associated with the metastatic lymph node status (p = 0.006). CONCLUSION: Immunohistochemical analysis of ER might be used as an easy, simple and highly efficient marker to identify high-risk patients and may aid in the selection of patients for a more aggressive adjuvant therapy. Furthermore, patients that show a positive ER-alpha immunostaining can be selected for a specific anti-hormonal therapy, as already performed for breast cancer patients.

Inhibin/activin-betaE subunit in normal and malignant human cervical tissue and cervical cancer cell lines.

Inhibins are dimeric glycoproteins, composed of an alpha-subunit and one of two possible beta-subunits (betaA or betaB), with substantial roles in human reproduction and in endocrine-responsive tumours. Recently a novel beta subunit named betaE was described, although it is still unclear if normal or cancerous cervical epithelial cells as well as cervical cancer cell lines can synthesise the novel inhibin-betaE subunit. About 4 normal cervical tissue samples together with 10 specimens of well-differentiated squamous cervical cancer and adenocarcinoma of the cervix were immunohistochemical analyzed. Additionally, two cervical carcinoma cell lines (HeLa and CaSki) were analyzed by immunofluorescence and RT-PCR for the expression of this novel subunit. We demonstrated for the first time an immunolabelling of the inhibin-betaE subunit in normal and malignant cervical tissue, as well as cervical cancer cells. Although the physiological role is still quite unclear in cervical tissue, inhibin-betaE might play important roles in carcinogenesis. Moreover, the synthesis of this subunit in cervical carcinoma cell lines of squamous and glandular epithelial origins also allows the use of these cell lines in elucidating its functions in cervical cancer pathogenesis. However, since the expression of the inhibin-betaE is minimal in HeLa cells as assessed by immunofluorescence and RT-PCR, the CaSki cell line might be a better model for further functional experiments regarding cervical cancer pathogenesis.

The human endometrium expresses the glycoprotein mucin-1 and shows positive correlation for Thomsen-Friedenreich epitope expression and galectin-1 binding.

Mucin 1 (MUC1) is a glycoprotein in human endometrium and is abundant at the luminal epithelial surface in the receptive phase. It has a highly glycosylated ecto-domain that contains keratan sulfate chains, that disappears at the time of implantation. In addition, the glycoforms on MUC1 differ in fertile and infertile women. Therefore the aims of this study were investigations on glycosylation of MUC1 with the Thomsen-Friedenreich (TF) epitope on normal human endometrium throughout the menstrual cycle and binding of galectin-1 on the TF epitope in the endometrium and the expression of galectin-1 on the human oocyte. Human endometrial tissue was obtained from 54 premenopausal patients and was immunohistochemically analyzed with monoclonal antibodies against MUC1, TF epitope, galectin-1, and biotinylated galectin-1. In addition, human oocytes were analyzed for TF, galectin-1 expression, and galectin-1 binding. We identified a significant upregulation of MUC1 and TF epitope and, in addition, galectin-1 binding in glandular epithelium and epithelial apical surface tissue from proliferative to secretory phase. With double staining experiments, we identified a coexpression of TF and MUC1 in the early secretory phase and galectin-1 binding to TF during the same period of time. In addition we identified TF epitope and galectin-1 expression plus binding on the human oocyte and irregularly fertilized oocytes. Upregulation of TF epitope on the glandular epithelium and epithelial apical surface tissue in the secretory phase and binding of galectin-1 at the same time show the possibility of galectin-1-mediated trophectoderm binding to the endometrium within the window of implantation.

Development and characterisation of an antibody for the immunohistochemical detection of inhibin/activin betaE (betaE) in normal human ovarian and placental tissue.

Inhibin/activin subunits are homologues to each other and belong to the transforming growth factor-beta (TGF-beta) family of proteins. These proteins have been demonstrated to be disulphide-linked dimers, which have a common alpha-subunit but just one of two beta-subunits, differentiated in inhibin A (alpha-betaA) and in inhibin B (alpha-betaB). Recently, an additional beta-subunit has been identified, determined as betaE and being primarily synthesized in liver tissue. However, since no antibody against the betaE subunit is commercially available, limited data on histological immunodistribution of this inhibin subunit in gynaecological organs exist. Therefore, the aims of the present study were the synthesis and evaluation of a specific antibody against the inhibin-betaE subunit. In this study, we describe the characterisation of a polyclonal antibody against the inhibin-betaE subunit. This antibody demonstrated a specific reaction in both western blot analysis and immunohistochemistry. Moreover, we demonstrated positive immunolabelling in normal human ovary and placenta. The role of this novel subunit is intriguing, especially within the view that the other inhibin/activin subunits might have substantial functions in human reproduction and carcinogenesis. However, the function of this subunit in humans remains still unclear and warrants further research.

Invasive hydatidiform mole: immunohistochemical labelling of inhibin/activin subunits, Ki67, p53 and glycodelin A in a rare case.

Invasive trophoblastic mole is an extremely rare condition. Its early recognition is essential since it can transform into an invasive type of tumour. Immunohistochemistry was performed with monoclonal antibodies against inhibin-alpha, -betaA and -betaB, Ki67, p53 and glycodelin A in a rare case of accidentally diagnosed invasive trophoblastic mole. There was labelling of the inhibin/activin subunits, Ki67 and p53, while glycodelin A showed minimal immunopositivity. Therefore, since the pathological diagnosis of an invasive mole is difficult, the immunohistochemical detection of inhibin/activin subunits, Ki67, p53 and glycodelin A might be additional useful tumour markers.

Inhibin-alpha subunit is an independent prognostic parameter in human endometrial carcinomas: analysis of inhibin/activin-alpha, -betaA and -betaB subunits in 302 cases.

Inhibins are dimeric glycoproteins, composed of an alpha-subunit (inhibin-alpha) and one of two possible beta-subunits (betaA or betaB), with substantial roles in human reproduction and in endocrine-responsive tumours. The aims of this study were to determine the distribution of inhibin-alpha, -betaA and -betaB subunits in malignant human endometrial tissue and the assessment of an association with specific clinicopathologic tumour features and clinical outcome. A series of 302 endometrial cancer tissue samples were immunohistochemically analysed with monoclonal antibodies against inhibin subunits. The inhibin-alpha subunit showed a significant association with histological grading, surgical staging, lymph node status and diabetes in patients with endometrial cancer. Interestingly, loss of inhibin-alpha expression resulted in a poorer survival of endometrial cancer patients. Additionally, survival analysis demonstrated that inhibin-alpha immunoreactivity was an independent prognostic factor for progression-free survival, cause-specific survival as well as for overall survival. In contrast, although inhibin-betaA- and -betaB subunits showed a significant association between endometrial histological subtypes and histological grading, both subunits were not found to be associated with survival in endometrial cancer patients. Therefore, inhibin-alpha immunostaining might be used as a simple and efficient marker to identify high-risk patients leading to the selection of patients for an aggressive adjuvant therapy.

The metastasis-associated gene MTA1 is upregulated in advanced ovarian cancer, represses ERbeta, and enhances expression of oncogenic cytokine GRO.

The metastasis-associated genes MTA1 and MTA3 are transcriptional repressors with potential effects on cancer. We analyzed the expression of MTA1, MTA3, ERalpha, ERbeta and E-cadherin in a total of 115 paraffin-embedded ovarian cancer tissues with respect to cancer staging and FIGO grading. Expression of MTA1, but not that of MTA3, was found to be significantly enhanced in ovarian cancer tissues with advanced cancer stages and higher FIGO grading, indicating an important role of MTA1 in the progression of ovarian cancer. To get further insights into the function of MTA1 in ovarian cancer, MTA1-overexpressing cancer cell clones were generated. In vitro, overexpression of exogenous MTA1 in OVCAR-3 cells had no effect on cell proliferation but enhanced the ability of anchorage-independent growth in soft agar colony formation assays. MTA1 overexpression resulted in downregulation of E-cadherin and MTA3 expression and enhanced expression of the transcriptional repressors SNAIL and SLUG. MTA1 further reduced ERbeta expression in vitro and inversely correlated with ERbeta expression in vivo. Screening for the expression of angiogenic cytokines expressed by ovarian cancer cells revealed MTA1-mediated upregulation of the oncogenic and angiogenic cytokine GRO (growth-regulated oncogene, CXCL1). Thus, in ovarian cancer, MTA1 expression directly and indirectly regulates the expression of several cancer-promoting as well as metastasis-facilitating factors, indicating an important role for MTA1 expression during ovarian cancer progression.

Prognostic significance of oestrogen receptor alpha (ERalpha) and beta (ERbeta), progesterone receptor A (PR-A) and B (PR-B) in endometrial carcinomas.

The expression of the classic steroid receptors ERalpha and PR-A has been correlated with stage, histological grade and survival in endometrial cancer. Endometrial cancer samples (293) were immunohistochemically analysed with monoclonal antibodies against the four steroid receptors. The loss of ERalpha, PR-A and PR-B resulted in a poorer survival in endometrial cancer patients, while ERbeta expression did not demonstrate any correlations with several analysed clinicopathological characteristics and did not affect survival. Additionally, multivariate survival analysis demonstrated that PR-B was a significant independent prognostic factor for cause-specific survival. In contrast, although ERalpha and PR-A showed a significant association between different endometrial histological subtypes and grading, both receptors were not independent factors with survival in endometrial carcinoma patients. Therefore, the PR-B immunostaining might be used as an easy, simple and highly efficient marker to identify high-risk patients and may aid in the selection of patients for a more aggressive adjuvant therapy.

Expression of E-cadherin in human ductal breast cancer carcinoma in situ, invasive carcinomas, their lymph node metastases, their distant metastases, carcinomas with recurrence and in recurrence.

BACKGROUND: Breast cancer cells can invade and generate metastasis via either lymphatic or blood vessels. E-cadherin mediates tumor cell-cell adhesion. Partial or complete loss of E-cadherin expression correlates with poor prognosis in breast cancer patients. In this study, the expression of E-cadherin was examined in mammary ductal carcinoma in situ, invasive breast carcinomas without metastasis, invasive carcinomas with their lymph node and distant metastases and invasive carcinomas with local recurrence in breast cancer tissue. MATERIALS AND METHODS: Paraffin-embedded slides of carcinoma in situ (8 DCIS), invasive carcinomas without lymph node metastases (9 invasive ductal carcinomas), invasive carcinomas (7 invasive ductal carcinomas) with corresponding lymph node metastases, invasive carcinomas (8 invasive ductal carcinomas) with corresponding recurrence and invasive carcinomas (5 invasive ductal carcinomas) with corresponding distant metastases were investigated for E-cadherin expression. Tissue slides were incubated with monoclonal antibodies against E-cadherin and stained with the ABC-elite kit. Staining intensities were analyzed by using a semi-quantitative score. RESULTS: A strong expression of E-cadherin in carcinoma in situ was demonstrated. Expression of E-cadherin was moderate in invasive carcinomas without metastases. However, very weak expression of E-cadherin in primary carcinoma with lymph node metastases was detected. E-cadherin expression was elevated in lymph node metastases compared to the primary tumor. CONCLUSION: Analysis of a tumor antigen involved in adhesion of breast cancer cells showed that there are significant differences of expression of E-cadherin between primary breast cancer cells and their metastases. Evaluation of this marker involved in cell adhesion could be a useful method for evaluating the metastatic risk in breast cancer patients.

Mucin 1, Thomsen-Friedenreich expression and galectin-1 binding in endometrioid adenocarcinoma: an immunohistochemical analysis.

BACKGROUND AND AIM: Altered mucin 1 (MUC1) secretion patterns have been implicated in several cancerous conditions including gastric, colorectal and breast carcinomas. Additionally, an association between the expression of MUC1, Thomsen-Friedenreich (TF) antigen, and binding of gal-1 (gal-1) has been proposed. Therefore, the aims of this study were to determine the frequency and tissue distribution of MUC1, TF and gal-1 binding in endometrioid adenocarcinomas. MATERIALS AND METHODS: Endometrial carcinomas diagnosed with only one histological tumor form (endometrioid adenocarcinomas) were obtained from 70 patients and classified according to the WHO grading system (G1 = 50; G2 = 12; G3 = 8). An immunohistochemical analysis was performed with specific antibodies against MUC1 and TF and in addition with biotinylated gal-1, followed by a semiquantitative evaluation and statistical analysis (chi2 test and Spearman's correlation coefficient). RESULTS: MUC1, TF and gal-1 were observed in human endometrioid adenocarcinomas. The MUC1 and gal-1 immunoreaction increased from G1 to G3, while TF demonstrated a lower intensity in G3 compared to G1, although with no statistical significance. However TF showed a significant correlation with MUC1 (p = 0.019) in G1 and G2 endometrioid adeno-carcinomas, with no observed correlation in G3 tumors. MUC1 and TF demonstrated a significant (p = 0.006 and p = 0.046, respectively) down-regulation in surgically staged FIGO III/IV compared to FIGO I/II. Gal-1 binding was up-regulated in FIGO III/IV although with no statistical significance. Interestingly, there was an association between gal-1 binding and lymphangiosis (p = 0.008). CONCLUSION: An immuno-histochemical expression of MUC1 and TF and gal-1 binding was demonstrated in human endometrioid adenocarcinomas. Although no significant expression patterns could be demonstrated within different nuclear grading, TF and MUC1 showed a significant correlation in G1/G2 tumors. Therefore, MUC1 and TF might be associated with endometrial malignant transformation. Additionally, MUC1 and TF were down-regulated in stage III/IV tumors, while a higher binding of gal-1 was observed in stage III/IV tumors, suggesting a substantial role of this antigen in endometrial carcinogenesis. Gal-1 binding was associated with lymphangiosis, which is thought to be a poor prognostic marker in endometrial adenocarcinomas. Therefore, MUC1, TF and galectin might have important roles in endometrial pathogenesis and malignant transformation. However, their utilization as specific tumor markers remains unclear and further studies are warranted.

Expression of inhibins in the endometrial carcinoma cell line RL-95-2 after stimulation with cortisol and estradiol.

UNLABELLED: Inhibins (INH) are dimeric glycoproteins composed of an alpha-subunit (INH-alpha) and one of two possible beta-subunits (INH-betaA or -betaB), with substantial roles in human reproduction and in endocrine-responsive tumours. The aim of the present study was the determination of the frequency and tissue distribution patterns of the inhibin/activin subunits in endometrial carcinoma cells of the cell line RL-95-2 after stimulation with estradiol and cortisol compared to unstimulated controls. MATERIALS AND METHODS: Cells of the endometrial carcinoma cell line RL-95-2 were grown on quadriperm tissue slides and incubated with different concentrations (0.1 and 0.01 micromol/ml) of estradiol or cortisol. Expression of INH-alpha, betaA and betaB was analysed by immunocytochemistry with specific monoclonal antibodies directed against the inhibin subunits. RESULTS: Expression of INH-alpha and -betaB was higher in cortisol-stimulated RL-95-2 cells, whereas INH-betaA expression was lower. In contrast to these, INH-betaB expression was increased by estradiol while INH-alpha and -betaA were unchanged under estradiol treatment. CONCLUSION: Expression of INH-subunits in RL-95-2 cells was described. Cortisol and estradiol showed an influence on INH expression. The RL-95-2 cell line could act as a useful model for the investigation of INH regulation, particularly for endometrial cancer.

The immunohistochemical expression of the inhibin/activin subunits is up-regulated by interferon-beta1a in Ishikawa cell line.

BACKGROUND: Inhibins are dimeric glycoproteins belonging to the TGF-beta1 family and are composed of an a-subunit (INH-alpha) and one of two possible beta-subunits (betaA or betaB; INH-betaA and INH-betaB). They have a substantial function in human reproduction and also seem to play an important role in endocrine-responsive tumors. Interestingly, there is an association between interferon and TGF-beta expression. However, this relationship has not been assessed in endometrial tissue regarding inhibin/activin expression. Therefore, the aim of this study was to determine expression changes of inhibin/activin subunits in the endometrial Ishikawa carcinoma cell line after stimulation with interferon-beta1a. MATERIALS AND METHODS: The Ishikawa cell line was cultured until confluence was observed (after 2 days). After adding interferon-beta1a (1000 IE/ml) Ishikawa cells were immunohistochemical analyzed for INH-alpha, INH-betaA and INH-betaB subunits. Experiments were performed in triplicates. The immunohistochemical expression was analyzed with a semiquantitative score (IRS) and statistical analysis was performed. RESULTS: Immunohistochemical reaction with INH-alpha could not be demonstrated in unstimulated cells, while it was significantly up-regulated in interferon-stimulated cells (p < 0.02). INH-betaA and INH-betaB were primarily observed during the mitotic phases of unstimulated cells. After stimulation their expression was significantly higher (p < 0.05 each) compared to controls and could be observed not only during mitotic phases but also in nonmitotic cells. CONCLUSION: For the first time, we demonstrated a functional relationship between interferon and inhibin/activin subunits. The expression of INH-alpha, INH-betaA and INH-betaB were immunohistochemical significantly up-regulated in the Ishikawa endometrial cell line after stimulation with interferon-beta1a. Since INH-alpha is thought to be tumor suppressive in the mouse model, interferon-beta1a might activate its gene. It remains to be clarified if this effect can be used as therapeutic options in endometrial carcinomas.

Inhibin/activin subunits are immunohistochemically expressed in complete and partial hydatidiform moles.

BACKGROUND AND AIM: Inhibins are dimeric glycoproteins, belonging to the transforming growth factor beta (TGF-beta) family, composed of an alpha-subunit (INH-alpha) and one of two possible beta-subunits (betaA or betaB). Additionally two further beta-subunits (betaC and betaE) have been cloned, although their function remains still quite unclear. The detection by immunohistochemistry of inhibin/activin subunits has been proposed as a useful marker of trophoblastic diseases. Interestingly, a complete mole cannot be easily differentiated from a partial mole. Therefore, the aim of this study was to determine expression changes of the five inhibin/activin subunits in partial and complete moles. MATERIALS AND METHODS: Histologically diagnosed complete (n = 6) and partial (n = 3) hydatidiform moles were immunohistochemical analyzed for INH-alpha, INH-betaA, INH-betaB, INH-betaC and INH-betaE subunits. The immunohistochemical reaction in intermediate trophoblast was analyzed with a semiquantitative score (IRS) and statistical analysis was performed. RESULTS: Immuno-histochemical reaction with INH-alpha, INH-betaA, INH-betaB, INH-betaC and INH-betaE subunits was demonstrated in hydatidiform moles. The INH-betaA and INH-betaB expression was significantly higher in complete compared to partial moles (p < 0.05 each), while INH-alpha, INH-betaC and INH-betaE did not demonstrate any statistically significant differences. CONCLUSION: We demonstrated an immunohistochemical expression of all five inhibin/activin subunits in partial and complete hydatidiform moles. The expression of INH-betaA and INH-betaB determined immunohistochemically was significantly up-regulated in complete moles, suggesting the utilization of these antibodies as diagnostic differentiation markers between complete and partial moles.

Expression of HPV, steroid receptors (ERalpha, ERbeta, PR-A and PR-B) and inhibin/activin subunits (alpha, betaA and betaB) in adenosquamous endometrial carcinoma.

AIM: The aim of the study was to determine possible pathogenetic factors and molecules which may be used as tumor markers of adenosquamous endometrial carcinoma. MATERIALS AND METHODS: Eight adenosquamous endometrial carcinomas were immunohistochemically tested with specific monoclonal antibodies for HPV (polyclonal anti-HPV and monoclonal anti-HPV 18), estrogen receptors ER-alpha and ER-beta, progesterone receptors PR-A and PR-B and the inhibin/activin subunits inhibin-alpha, -betaA and -betaB. RESULTS: HPV 18 and the polyclonal HPV antibody was detected in all adenosquamous endometrial carcinomas, both in the endometrioid (n = 7/8) and squamous (n = 8/8) parts of the tumor. Neither ER-alpha or ER-beta were detectable in any tumor, in contrast to PR-A and PR-B which were detected in about half of these tumors (PR-A: n = 5/8 and PR-B: n = 2/8). Inhibin-alpha and -betaB were not detected, while inhibin-betaA was expressed in all adenosquamous carcinomas. CONCLUSION: The carcinogenesis of adenosquamous endometrial carcinomas was associated with HPV infection. Adenosquamous endometrial carcinomas seem not to be controlled by estrogens. The absence of the expression of the inhibin-alpha subunit suggests a tumor-suppressive function in adenosquamous endometrial tumors. The absence of the expression of the inhibin-betaB subunit, which is probably a marker of differentiation, points to the malignancy of these tumors. The other inhibin subunit, inhibin-betaA, was expressed in all adenosquamous tumors. It remains to be clarified if these parameters can be used as tumor markers.