Recent studies have been reported the roles of the estrogen receptor (ER), progesterone receptor (PR) and p53 in the development of a pelvic organ prolapse (POP). The pathogenesis of stress urinary incontinence (SUI) is related to that of POP in the weakness of pelvic support. Therefore, this study was carried out to assess the relationship between ER, PR, p53 and the development of SUI, and to elucidate the biomolecular pathophysiology of SUI. The periurethral fascia was obtained from 6 menopausal patients diagnosed with SUI and 10 menopausal patients without SUI who visited the Department of Obstetrics and Gynecology, Severance Hospital, Seoul, Korea. The relative ER, PR and p53 protein levels in the periurethral fascia were obtained by western blot analysis and densitometry. A Mann-Whitney U test was used for statistical analysis (p< 0.05). The mean age (+/-SD) of the 16 patients was 59.0 +/-5.5 years (range, 50-74 years). The mean body mass index was 25.2 +/-2.7 kg/m2 (21.8 - 30.8) and the average number of vaginal deliveries was 2.8 +/-1.9 (1.0 - 9.0). The ER level (0.33 +/-0.17 vs. 1.86 +/-0.83, p= 0.02) and the p53 level (1.25 +/-0.67 vs. 4.71 +/-2.40, p= 0.01) were lower in the experimental group. However, the PR level of the two groups were similar (0.64 +/-0.13 vs. 0.48 +/-0.33, p=0.56). The p53 and ER levels were significant lower in the study group. This suggests that p53 and ER might be important factors in the development of SUI. Further prospective studies about the association of ER, p53 and SUI will be needed to elucidate the molecular pathogenesis of SUI.
Aged ; Female ; Humans ; Middle Aged ; Protein p53/analysis/*physiology ; Receptors, Estrogen/analysis/*physiology ; Receptors, Progesterone/analysis/*physiology ; Urinary Incontinence, Stress/*etiology

Estrogen-related receptor a (ERRalpha) is a key regulator for energy metabolism and adipogenesis. However, its role in lipolysis is unknown. To study the function of ERRalpha in lipolysis, primary cultured differentiated porcine adipocytes were treated by a specific inverse agonist XCT790 or infected with adenoviral vector expressed ERRalpha for 48 h, in the absence and/or presence of specific protein kinase A (PKA) inhibitor or extracellular signal-related kinase (ERK) inhibitor. Then, we measured the triglyceride (TG) content and the glycerol release into the culture media to analysis the effect of ERRalpha on lipolysis; Further, we analyzed the expression of PPARgamma, perilipin A, p-perilipin A, HSL and ATGL with Western blotting. Here, we found that ERRalpha significantly increased adipocytes differentiation, TG accumulation and glycerol release. Separately or simultaneously block the PKA and ERK pathway do not significantly altered the effect of ERRalpha on glycerol release. ERRalpha significantly up-regulated the proteins expression of PPARgamma, perilipin A, HSL and ATGL, while the p-perilipin A protein level was not significantly changed. These findings imply that ERRalpha could increase lipolysis via up-regulating HSL and ATGL, thereby to supply more FFA as substrate for a larger turnover of cellular triglyceride pool during adipocytes differentiation.
Adipocytes ; cytology ; metabolism ; Animals ; Animals, Newborn ; Cells, Cultured ; Glycerol ; analysis ; Lipase ; metabolism ; Lipolysis ; physiology ; Receptors, Estrogen ; metabolism ; physiology ; Sterol Esterase ; metabolism ; Swine ; Triglycerides ; analysis

Objective: To investigate the expression of the G-protein coupled estrogen receptor (GPER) in the testis of the male mouse with kidney yin or kidney yang deficiency and its influence on the reproductive function of the mouse. METHODS: We randomized 30 six-week-old male Kunming mice into three groups of equal number: kidney yang deficiency, kidney yin deficiency, and normal control, and established the models of kidney yang deficiency and kidney yin deficiency by peritoneal injection of hydrocortisone at 50 mg/kg for 5 days and 25 mg/kg for 10 days, respectively. We observed the behavioral changes of the mice using the elevated plus-maze, exhaustive swimming and field experiment, examined the semen quality with the automatic sperm quality analyzer, calculated the average number of the offspring, measured the serum testosterone (T) and estradiol (E2) levels and T/E2 ratio by Roche electrochemiluminescence assay, and determined the localization and expression of GPER in the testis by immunohistochemistry and immunofluorescence staining. RESULTS: Compared with the mice with kidney yin deficiency, those with kidney yang deficiency showed remarkably fewer entries into the open arm and central area (P <0.05) and shorter time of exhaustive swimming (P <0.05), but no statistically significant difference in the time spent in the open arm or the central area (P >0.05); the latter group also exhibited significant decreases in the epididymal sperm count (［7.27 ± 1.30］ vs ［3.05 ± 1.06］ ×108/g, P <0.01), sperm motility (［54.15 ± 13.52］ vs ［51.57 ± 8.75］ %, P <0.01) and average number of the offspring (6.46 vs 4.33, P <0.05), a slight increase in the rate of morphologically abnormal sperm (［13.42 ± 2.32］ vs ［15.39 ± 2.48］ %, P >0.05), and markedly reduced serum T (［24.96 ± 6.18］ vs ［16.72 ± 5.92］ ng/dl,P <0.05), E2 (［19.81 ± 4.01］ vs ［15.24 ± 1.11］ pg/ml,P <0.05) and T/E2 ratio (1.41 vs 1.25, P <0.05). The expression of GPER was found in the cytoplasm of the Leydig cells, negative in the nuclei and cell membrane, significantly higher in the kidney yang than in the kidney yin deficiency group (P <0.05). CONCLUSIONS The numbers of sperm and offspring decreased while the percentage of morphologically abnormal sperm increased in both the kidney yang and kidney yin deficiency mice, even more significantly in the former, which might be associated with the up-regulated expression of GPER in the testis of the mouse with kidney yang deficiency and consequently the reduced serum T level and T/E2 ratio.
Animals ; Drugs, Chinese Herbal ; Kidney Diseases ; metabolism ; Male ; Mice ; Random Allocation ; Receptors, Estrogen ; metabolism ; Receptors, G-Protein-Coupled ; metabolism ; Reproduction ; physiology ; Semen Analysis ; Testis ; metabolism ; Yang Deficiency ; metabolism ; Yin Deficiency ; metabolism

AIMTo investigate the effect of androgen on the proliferation, differentiation and regression of canine prostatic stromal cells in vivo and human stromal cells in vitro.

METHODSTwenty-two dogs, including 15 normal prostate dogs and 7 prostatic hyperplasia dogs, had their serum concentration of testosterone and estrodiol determined by radioimmunoassay before and after castration. The expression of androgen receptor (AR) and estrogen receptor (ER) in the prostate were analysed by immunohistochemistry and semi-quantitative RT-PCR before and after castration. Light microscopy, transmission electron microscopy and TUNEL assay were carried out successively before and after castration to evaluate the prostatic histomorphology. In vitro serum-free cell cultures from human prostatic stroma were established and exposed to dihydrotestosterone (DHT). The proliferation of the cell culture was detected by MTT assay. The expression of TGFbgr, bFGF, AR, and smooth muscle cell (SMC) specific proteins (myosin and/or smoothelin) were detected using immunohistochemistry and RT-PCR. The differentiation from fibroblasts to smooth muscle cells was deduced by measuring the expression of SMC specific proteins.

RESULTSBefore castration, the serum concentrations of testosterone and estrodiol were not statistically different between normal and hyperplasia groups. Following castration, the serum concentration of testosterone decreased rapidly in 2 days, and the concentration of estrodiol had no significant change compared with the pre-castration data. In the prostate, AR was presented in both the epithelial and stromal cells and the AR mRNA level was higher in hyperplasia than in normal prostate tissues (P<0.05). While ER predominantly existed in the prostate stromal cells and the ER mRNA had no difference between the hyperplasia and the normal group. Within the early phase of castration (

CONCLUSIONThe whole prostate gland is an androgen-sensitive organ with both the epithelium and stroma under the control of androgen. Androgen may direct the proliferation, differentiation and regression of stromal cells by regulating the expression of TGFbgr, bFGF, AR and smooth muscle cell specific proteins.

Animals ; Biomarkers ; Cell Differentiation ; drug effects ; physiology ; Cell Division ; drug effects ; physiology ; Cells, Cultured ; Dihydrotestosterone ; pharmacology ; Dogs ; Estradiol ; blood ; Fibroblast Growth Factor 2 ; genetics ; pharmacology ; Gene Expression ; Humans ; Male ; Muscle, Smooth ; cytology ; physiology ; Orchiectomy ; Prostate ; cytology ; physiology ; Prostatic Hyperplasia ; physiopathology ; RNA, Messenger ; analysis ; Receptors, Androgen ; genetics ; Receptors, Estrogen ; genetics ; Stromal Cells ; cytology ; physiology ; Testosterone ; blood ; Transforming Growth Factor beta ; genetics ; pharmacology

OBJECTIVETo summarize the morphological features of basal-like subtype of invasive breast carcinoma (BLSIBC), and to look for diagnostic clues for its recognition.

METHODSImmunohistochemistry was performed in 109 cases of invasive ductal carcinoma, with CK5/6, CK14, CK8/ 18, 34betaE12, calponin, p63, CD10, ER, PR and c-erbB-2 monoclonal antibodies. Five subtypes were classified according to immunophenotypes: luminal A subtype (ER+/HER2-), luminal B subtype (ER+/ HER2+), normal breast-like subtype (ER/HER2-), HER2-overexpressing subtype and BLSIBC which was identified with at least one kind of basal-like cytokeratins or markers of myoepithelium and ER/HER2. The microscopic features of basal-like subtype were also analyzed.

RESULTSThe number of luminal A case was 48 (44.0%), luminal B 15 (13.8%), HER2 over-expressing 15 (13.6%), normal breast-like 10 (9.1%), basal-like subtype 19 (17.4%). Besides, the other two cases expressed c-erbB-2 or/and ER plus markers for myoepithelium, thus were excluded from all the five mentioned subtypes. Of the 19 basal-like subtype, CK5/6 was expressed in 16 cases, CK8/18 in 17 cases, CK14 in 11 cases, 34betaE12 in 18 cases, p63 in 5 cases, CD10 in 6 cases, and calponin in 1 case. The diameter of the BLSIBC cases was 1.2-7 cm (averagely 3.9 cm) , and in 6 cases, the tumor diameter was >5 cm. Only one case displayed extensive in situ component, 9 cases were grade 2, and 9 cases were grade 3. Compared to non basal subtype, there were significantly more high grade cases (P <0.01). The morphological features of basal-like subtype were summarized as the followings: pushing margin (13 cases), lymphocytic tissue hyperplasia (18 cases), nest or sheet arrangement (18 cases), nucleus grade 3 and scattered giant or bizarre nuclei (17 cases), syncytial growth (7 cases), and comedo-like necrosis (17 cases). The frequency of these features were significantly more common than non basal subtype (P <0.01).

CONCLUSIONThe morphologic diagnostic features of BLSIBC are pushing margins, lymphocyte infiltration, comedo-like necrosis, gigantic cell and syncytial growth.

Biomarkers, Tumor ; analysis ; Breast Neoplasms ; diagnosis ; pathology ; Carcinoma, Basal Cell ; pathology ; Female ; Genes, erbB-2 ; physiology ; Humans ; Immunohistochemistry ; Keratin-5 ; analysis ; Magnetic Resonance Imaging ; Male ; Mammography ; instrumentation ; methods ; Neoplasm Invasiveness ; physiopathology ; Prognosis ; Receptor, Epidermal Growth Factor ; genetics ; Receptor, ErbB-2 ; analysis ; genetics ; Receptors, Estrogen ; analysis ; Receptors, Progesterone ; analysis ; Ultrasonography ; methods

BACKGROUNDEstrogen receptor (ER) is a very important biomarker of breast cancer. ER deletion has been consistently associated with tumor progression, recurrence, metastasis and poor prognosis, but the biological mechanism is still unclear. ER negative breast cancer expresses high levels of interleukin-8 (IL-8). ER expression can downregulate IL-8 promotor activity. As a multifunctional cytokine, IL-8 has many important biological activities in tumor genesis and development. With the goal of investigating the role of IL-8 in ER-negative breast cancer progression, we applied RNA interference technology to specifically knockdown the IL-8 expression in ER-negative breast cancer cell line MDA-MB-231.

METHODSInterfering pRNA-IL-8 and the control was transfected into ER (-) MDA-MB-231. The proliferation, cell apotosis, and invasive ability were recorded in transfected, untransfected and negative transfected cells. These cells were injected into nude mice to assess tumorigenicity, proliferation, metastasis and microvessel density (MVD).

RESULTSIn vitro, decreased expression of IL-8 was associated with reduced cell invasion (P < 0.001), but had no effect on cell proliferation (P > 0.05). In vivo, neutrophils infiltration was significantly inhibited in pRNA-IL-8 transfected cells compared with untransfected and negatively transfected cells (P = 0.001, P < 0.001). Less metastasis was found in transfected cells compared with negatively transfected cells (0% vs 80%, P = 0.048). Nevertheless, we observed less MVD in transfected cells compared with control in nude mice (P < 0.001).

CONCLUSIONSIL-8 inhibits ER-negative breast cancer cell growth and promotes its metastasis in vivo, which may be correlated with neutrophils infiltration induced by IL-8.

Apoptosis ; Breast Neoplasms ; blood supply ; chemistry ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Disease Progression ; Female ; Humans ; Interleukin-8 ; antagonists & inhibitors ; physiology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neutrophil Infiltration ; RNA, Small Interfering ; pharmacology ; Receptors, Estrogen ; analysis

OBJECTIVETo identify the effect of interleukin-8 in cell progression and invasion of human breast cancer.

METHODSHuman cytokine antibody arrays were applied to screen a panel of cytokine expression from 11 human breast cancer cell lines, and the mechanism of identified key factors involved in breast cancer progression was studied.

RESULTSProfiling of cytokine expression showed the expression of interleukin-8 was related to estrogen receptor status, metastasis and vimentin status in the 11 human breast cancer cell lines. Elevated expression of interleukin-8 in breast cancer cells had positive correlation with breast cancer invasion. Neutralization of antibody against interleukin-8 specifically blocked interleukin-8-mediated cell invasion. However, anti-interleukin-8 antibody did not influence the proliferation of breast cancer cells.

CONCLUSIONInterleukin-8 may be the key factor involved in human breast cancer progression and invasion, and play an important role in cell invasion of breast cancer.

Breast Neoplasms ; metabolism ; pathology ; Cell Proliferation ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunohistochemistry ; Interleukin-8 ; biosynthesis ; physiology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Protein Array Analysis ; Receptors, Estrogen ; metabolism ; Tumor Cells, Cultured ; Tumor Suppressor Protein p53 ; metabolism