Functional evidence for two distinct mechanisms of action of progesterone and selective progesterone receptor modulator on uterine leiomyomas.

OBJECTIVE: To study the specific mechanisms through which progesterone and selective progesterone receptor modulators impact the growth, synthesis, and accumulation of the extracellular matrix in uterine leiomyomas. DESIGN: Laboratory study. SETTING: Academic Research Institutions. PATIENTS (S): This study involved reproductive-age women diagnosed with infertility associated uterine leiomyomas who underwent myomectomy either after selective progesterone receptor modulator ulipristal acetate (UA) treatment or without any pharmacological pretreatment. Control samples included healthy myometrium tissue (n = 100). Specimens were obtained from the Department of Reproduction and Gynecological Endocrinology and Biobank, Medical University of Bialystok, Poland. INTERVENTIONS: Daily (5 mg/d) UA treated for 2 months (n = 100) and untreated (n = 150) patients with uterine leiomyomas or normal healthy myometrium (n = 100) tissue samples immediately after surgery were collected for transcriptional analysis and assessments. MAIN OUTCOME MEASURES: Progesterone-induced activation of the signaling pathways related to uterine leiomyomas extracellular matrix synthesis, deposition, and growth, as well as the expression profile of progesterone receptors in uterine leiomyomas, were assessed. RESULTS: The results indicated that progesterone activated the transforming growth factor-ß and SMAD3 signaling pathways and promoted proliferation, growth, and extracellular matrix remodeling in uterine leiomyomas by up-regulating SMAD3, transforming growth factor-ß (TGF-ß) receptor type 1 and II, Ras homolog A, vascular endothelial growth factor, or increasing the fibrosis-related gene collagen, type I, ɑ-1, and procollagen, type I, ɑ-1 production. In contrast, UA had inhibitory effects on these processes. The study also showed that both nuclear and membrane progesterone receptors play distinct roles in uterine leiomyoma pathobiology. CONCLUSIONS: We showed that both nuclear and membrane progesterone receptors were relevant in the treatment of uterine leiomyomas, especially when combined with selective progesterone receptor modulators. Novel therapeutic approaches combining selective progesterone receptor modulators with or without direct and indirect extracellular matrix targeting through selected specifically TGF-ß and SMAD3 (SMAD3, TGF-ß receptor types 1 and II, Ras homolog A, vascular endothelial growth factor, collagen, type I, ɑ-1) signaling pathways could therefore be a treatment option for uterine leiomyomas.

Potential of mucoadhesive chitosan glutamate microparticles as microbicide carriers - antiherpes activity and penetration behavior across the human vaginal epithelium.

Chitosan glutamate (gCS) spray-dried microparticles appear promising carriers to overcome challenges associated with vaginal microbicide delivery. This study aimed at elucidating the penetration and mucoadhesive behavior of developed gCS multiunit carriers with zidovudine (ZVD) as a model antiretroviral agent in contact with excised human vaginal epithelium followed with an examination of in vitro antiherpes activity in immortal human keratinocytes HaCaT and human vaginal epithelial cells VK2-E6/E7. Both ZVD dispersion and placebo microparticles served as controls. Microparticles displayed feasible (comparable to commercial vaginal product) mucoadhesive and mucoretention characteristics to isolated human vaginal tissue. Ex vivo penetration studies revealed that gCS increased the accumulation of active agent in the vaginal epithelium but surprisingly did not facilitate its penetration across human tissue. Finally, the obtained antiviral results demonstrated the potential of gCS as an antiherpes adjunctive, whose mode of action was related to blocking viral attachment.

Functional Expression of FSH Receptor in Endometriotic Lesions.

CONTEXT: FSH receptor (FSHR), besides being expressed in gonads, is also expressed in some extragonadal tissues at low levels. OBJECTIVE: We examined the functional expression of FSHR in different types of endometriotic lesions. DESIGN: Extensive studies were carried out to detect functional FSHR expression and FSH-stimulated estrogen production in ovarian endometriomas and recto-vaginal endometriotic nodules (RVEN). Normal endometrium, ovary, and myometrium tissues from nonpregnant cycling women served as controls. SETTINGS: This laboratory-based study was carried out on tissue specimens from patients with endometriosis and healthy donors. RESULTS: Endometriotic lesions and normal secretory-phase endometrium showed FSHR expression at both mRNA and protein level. RVEN and ovarian endometrioma demonstrated up-regulated CYP19A1, dependent on the activation of CYP19A1 proximal promoter II. Estrogen receptor-ß (ESR2) expression was significantly increased in RVEN vs normal endometrium. Recombinant human FSH stimulation of RVEN explants significantly increased estradiol production and CYP19A1 and ESR2 expression. FSHR was up-regulated in recombinant human FSH-stimulated endometrial and decidualized stromal cells with increased CYP19A1 expression. CONCLUSIONS: We described a novel functional FSHR expression, where FSH-stimulated CYP19A1 expression and estrogen production in RVEN are demonstrated. This locally FSH-induced estrogen production may contribute to the pathology, development, progression, and severity of RVEN.

ERα and ERß expression in correlation with Ki-67, Bcl-2 and Bak in primary tumors and lymph node metastases of breast cancer: The effect of pre-operative chemotherapy.

This study aimed to assess the pre-operative chemotherapy impact on the relationship between estrogen receptor (ER) expression and markers of proliferation and apoptosis in primary and metastatic breast cancer. Immunohistochemical examinations were conducted on surgically removed ductal invasive breast cancers and their lymph node metastases in 135 patients. A total of 64 patients from this group underwent pre-operative chemotherapy and in 71 cases the surgery was performed without primary chemotherapy. A negative correlation between ERα and Ki-67 was found in primary tumors and lymph node metastases. A positive correlation was observed between ERα and Bcl-2. A positive correlation was also noted between ERß and Bak, suggesting that the two ERs were involved in the regulation of proteins responsible for the control of the apoptotic process. Assessment of the expression of the proteins conducted separately in primary tumors and lymph node metastases did not reveal a significant effect of pre-operative chemotherapy on the correlations of ERs with Ki-67, Bcl-2 and Bak. However, the analysis of the correlations between the receptor expression in primary tumors and Ki-67, Bcl-2 and Bak in lymph node metastases showed a statistically significant impact of pre-operative chemotherapy on the correlations of ERα and Bcl-2 with ERß and Bak, confirming involvement of the two ERs in the regulation of apoptosis during breast carcinogenesis.

The effect of chemotherapy on Ki-67, Bcl-2 and Bak expression in primary tumors and lymph node metastases of breast cancer.

The aim of this study was the analysis of Ki-67, Bcl-2 and Bak expression in primary tumor and axillary lymph node metastases of breast cancer as well as an attempt to assess preoperative chemotherapy influence on the mentioned markers with regard to changes in the morphological appearance of the primary tumor and its metastases. Immunohistochemical examinations of Ki-67, Bcl-2 and Bak expression were conducted on sections collected from 135 patients treated surgically on invasive ductal breast cancer. Sixty-four of these patients were administered preoperative chemotherapy, whilst on 71 patients the surgery was performed without initial chemotherapy. In the group of patients without preoperative chemotherapy positive correlation in Ki-67 and Bcl-2 expression between primary tumors and lymph node metastases (p<0.0001, r=0.707; p<0.0001, r=0.604, respectively) was observed. In the group of patients after chemotherapy positive correlation between primary tumors and lymph node metastases in case of Bcl-2 and Bak proteins (p<0.04, r=0.424; p<0.02, r=0.478, respectively) was observed. It was also found that preoperative chemotherapy has an influence on the expression of proteins connected with proliferation and apoptosis and thus, it can influence neoplastic process biology. It does not have any significant impact on the proapoptotic Bak protein expression either in primary tumor or in lymph node metastases of breast cancer. However, it is related to lower expression of antiapoptotic Bcl-2 protein (p<0.0005) and of Ki-67 proliferation marker (p<0.03) in primary tumors, which indirectly indicates a beneficial influence of preoperative chemotherapy on the primary tumor. Concurrently, the influence of neoadjuvant therapy on lymph node metastases seems to be relatively small, which can limit its effectiveness.

The effect of chemotherapy on status of estrogen receptors in primary tumors and lymph node metastases of human ductal breast cancer.

The aim of the study was the evaluation of ERalpha and ERbeta expression in primary tumors and lymph node metastases of breast cancer as well as the assessment of the influence of preoperative chemotherapy on these receptors with regard to changes in morphological appearance of primary tumors and their metastases. Immunohistochemical examinations were conducted on surgically removed ductal invasive breast cancers and their lymph node metastases of 135 patients. Seventy-one patients were spared preoperative chemotherapy which was administered to other 64 patients. Primary breast cancers with preoperative chemotherapy showed lower mean percentage of cells with a positive reaction to ERalpha and ERbeta as compared to primary tumors without preoperative chemotherapy. There were positive correlations among primary tumors and lymph node metastases regardless of preoperative chemotherapy applied. On the other hand, ERalpha and ERbeta expressions were negatively correlated in primary tumors without chemotherapy in contrast to primary tumors after chemotherapy. Furthermore, it was observed that preoperative chemotherapy was responsible for significantly less damage to lymph node metastases of breast cancer in comparison to primary tumors. In cases of such advanced damage of primary tumors that made determination of estrogen receptor expression impossible, their evaluation was performed on metastases to regional lymph nodes. Although preoperative chemotherapy did not severely impair estrogen receptor expression, presented changes of their distribution are a sufficient reason for simultaneous labeling of estrogen receptors in both primary tumors and metastases due to various sensitivity to chemotherapy of primary cancers in comparison with involved lymph nodes.

Connexins 26 and 43 correlate with Bak, but not with Bcl-2 protein in breast cancer.

Apoptosis is a process associated with development and progression of breast cancer. The association between connexins and programmed cell death has only been demonstrated in a few studies. The aim of the present study was the evaluation of Cx26 and Cx43 expression in breast cancer in correlation with Bcl-2 and Bak proteins as well as with selected clinicopathological features. Tissue samples from 71 women were examined by immunohistochemistry, using the streptavidin-biotin-peroxidase complex technique for Cx26, Cx43, Bak and Bcl-2. Cytoplasmic expression of Cx26 and Cx43 was detected in 34 (47.9%) and 61 (85.9%) of breast cancers, respectively. Bcl-2 and Bak expression was found in 59 (83%) and 50 (70%) of studied cases, respectively. We found a positive correlation between Cx26 and Bak expression (r=0.541, p<0.0001) as well as between Cx43 and Bak (r=0.589, p<0.0001), but not between the studied connexins and Bcl-2. The expression of Cx26 was not associated with age, tumor size, lymph node status or histological grade. However, we observed an association between Cx43 expression and histological grade G3 (p<0.04). Cytoplasmic localization of evaluated connexins supports the concept of alterations in connexins expression in breast cancer cells. The associations of evaluated connexins with Bak protein could suggest that connexins localized in the cytoplasm may participate in signaling apoptotic pathways.

Expression of insulin-like growth factor-I receptor, estrogen receptor alpha, Bcl-2 and Bax proteins in human breast cancer.

Disturbance in expression of estrogen receptors together with changing influence of growth factor receptors and apoptosis associated proteins plays a role in breast cancer development and progression. However, immunohistochemical detection and relationships among these proteins were not often considered in relation to breast cancer and a few evaluations of expression provided mismatching results and conclusions. Consequently, we examined by immunohistochemistry the expression of the insulin-like growth factor-I receptor (IGF-IR), estrogen receptor alpha (ERalpha) and apoptosis-associated proteins, Bcl-2 and Bax, in human primary breast cancer, as well as analyzing the relationships among these proteins. The positive immunostaining for IGF-IR, ERalpha, Bcl-2 and Bax was noted in 56, 63.8, 82.8 and 50% of tumors, respectively. We observed that IGF-IR negatively correlated with ERalpha in the group of all tumors and in axillary node negative cancer (p<0.03, p<0.05, respectively), but not in the subgroup of node positive cancer. Expression of ERalpha correlated positively with Bcl-2 and negatively with Bax proteins (p<0.0001, p<0.05, respectively). We did not note significant relationships between IGF-IR and Bcl-2, or IGF-IR and Bax proteins. We found that increased Bax expression was associated with positive lymph node status, pT2 stage and G3 grade of tumors. Knowledge about alterations in the IGF-IR expression and relations of the receptor to other biological factors could help in our understanding of breast cancer biology and the importance of the IGF-IR in cancer progression as well as in effective management of breast cancer.

[Angiogenesis in the female reproductive processes]. / Angiogeneza w procesach rozrodczych u kobiety.

Angiogenesis or the formation of new vessels out of pre-existing capillaries is a sequence of events that is fundamental to physiology of the female reproductive tract and also pathologic processes such as ovarian hyperstimulation syndrome. Many factors include vascular endothelial growth factor (VEGF), angiopoietins and others involved in the regulation of angiogenesis have been identified. There are some endocrine control mechanisms, which stimulate or inhibit the angiogenesis. The studies indicate that the normal processes of folliculogenesis, ovulation and corpus luteum function in the ovary and the control of menstruation and implantation in the endometrium are profoundly dependent on the angiogenesis. The rapid, controlled and cyclical nature of angiogenesis in the female reproductive tract suggests that interference with this process should provide a novel approach to manipulation of reproductive function.

[Comparative studies of K1-67 expression between the primary tumor and breast cancer metastases to regional lymph nodes]. / Badania porównawcze ekspresji Ki-67 w guzie pierwotnym i w przerzutach raka sutka do regionalnych wezlów chlonnych.

Breast cancer is often relatively slow growing, but at diagnosis about 40% of patients have regional spread to at least one axillary node. It has been shown that unrelated clones are in primary breast carcinomas. There is possibility that only a small subpopulation of the cells of the primary tumour (PT) metastasis. It has been shown that Ki-67 protein is a useful marker in histopathology, which is present during all active phases of the cell cycle and making possibility to assess growth fraction of tumour cells. The purpose of the study was to evaluate the expression of Ki-67 and comparison between the PTs and MRLNs as well as to estimate the relationships between Ki-67 and the chosen anatomoclinical features of the breast cancer. Immunohistochemical analyses for Ki-67 were performed on the PTs (69 cases without primary chemotherapy) and MRLNs (33 cases) of breast cancer. Increased expression of Ki-67 in PTs significantly correlated with pT2 stage of tumours (p < 0.05) and grade G3 (p < 0.04), but there was not relationship with lymph node status. Expression of Ki-67 positively correlated between PTs and MRLNs (p < 0.001). Comparison between PTs and matching MRLNs revealed that 23 (69.7%) cases showed a convergence between PTs and matching MRLNs with regard to negative or positive staining. We would like to emphasize the importance of studies concerning the proteins involved in proliferation in MRLNs, because knowledge about heterogeneity between PTs and MRLNs could shed light on tumour biology and may lead to development of more effective anti-cancer therapies.