Expression and role of hScrib in endometrium, endometriosis, and endometrial adenocarcinoma.

To explore the role of hScrib in the pathogenesis of endometriosis.This was a retrospective study of 240 women in our hospital between January 2014 and January 2017. The expression of hScrib in endometrium (EM), endometriosis (EMs), and endometrial adenocarcinoma (EC) was investigated, and compared the differences among them. Serum levels, protein expressions, localizations, and correlations of hScrib and E-cadherin were determined.The levels of serum soluble hScrib and E-cadherin were significantly highest in EC, followed by EMs, and healthy women (P < .05). hScrib protein content was opposite result in 3 tissues (P < .05), and was negatively correlated with r-AFS stage in EMs. The location changed from membrane to cytoplasm. Co-localization of hScrib with E-cadherin was found at extensive cell-cell boundaries in EMs.hScrib and E-cadherin may be as new diagnostic markers of endometriosis. Low expression of hScrib leads to the loss of cell polarity and stability. Also, hScrib may induce EMT through regulating E-cadherin, might play an important role in pathogenesis of endometriosis.

[Culture and identification of fibroblast from human endometriosis].

OBJECTIVE: To establish the model of cultivating and identifying fibroblast from human endometriosis (HEFC) in vitro. METHODS: The tissues of human endometriotic cysts of ovary were digested by collagenases I, II and IV The resulting cells were purified by centrifugation and differential adhesion. HEFC was identified by observing the morphologic changes under an inverted microscope and the expressions of vimentin, α-SMA (α-smooth muscle actin) and keratin were detected by immunocytochemistry. RESULTS: Immunohistochemical staining of vimentin was positive, α-SMA rarely positive and keratin completely negative in cultured fibroblasts. HEFC grew as a confluent monolayer of short fat fusiform, triangular, star-shaped and polygonal fiber-like cells. Furthermore HEFC could be well sub-cultured. CONCLUSION: Acquired fibroblast can be cultured in vitro stably. It is quite important to study the specificities of HEFC. Sufficient and reliable target cells may be obtained for studying the mechanisms of fibrosis and adhesion in endometriosis at the molecular level.

[Culture and identification of microvascular endothelial cells from human endometriosis].

OBJECTIVE: To establish the methods of isolating and culturing human ovarian endometriosis-derived microvascular endothelial cells (OEMEC). METHODS: The tissues of human endometriotic cyst of ovary were finely minced with scissors, then digested by collagenase I, II and trypsin-ethylene diamine tetraacetic acid (EDTA). The cells were purified by using centrifugation of 2000 r/min speed. OEMEC were identified by light microscope and transmission electron microscope observing CD(34), FVIII-Rag and Weibel-Palade in microvascular endothelial cells. RESULTS: The OEMEC grew as confluent monolayer like cobblestones under light microscope. CD(34) and FVIII-Rag were expressed strongly, and the percentages of CD(34) and FVIII-Rag positive cells were 91.4% and 92.5%. Weibel-Palade bodies could be observed under transmission electron microscope. The time of cell doubling proliferation was 4.5 days. CONCLUSION: The established system of isolating OEMEC would provide lab base for studying the mechanisms of angiogenesis in endometriosis lesions.

hScrib, a human homolog of Drosophila neoplastic tumor suppressor, is involved in the progress of endometrial cancer.

Recent studies have revealed that hScrib, the human homolog of Drosophila Scribble, is an apical-basal polarity determinant and an essential component of the adherens junction. In addition, hScrib has a critical role in the inhibition of cell proliferation through cell cycle progression. hScrib has been reported to be involved in the processes of many cancers, such as breast cancer, cervical cancer, colon carcinoma, etc.; however, the correlation between hScrib and endometrial cancer has not been identified. To address a possible role of hScrib in the development of endometrial cancer, we examined the localization and expression of hScrib in endometrial cancer. The present study demonstrated that decreased expression and changed localization of hScrib were associated with clinical stage, histopathological differentiation, and lymph node metastasis in endometrial cancer. hScrib might share an adherens junction with basolateral membrane partially by acting on E-cadherin in endometrial cancer. This evidence suggests that hScrib is involved in the development of endometrial cancer.

Interleukin-4 induces expression of eotaxin in endometriotic stromal cells.

OBJECTIVE: To study the relationship between eotaxin and interleukin-4 (IL-4) in the pathophysiology of endometriosis. DESIGN: Comparative and laboratory study. SETTING: University teaching hospital reproductive endocrinology and infertility practice. PATIENT(S): Ectopic endometrial tissues were collected from women with endometriosis. INTERVENTION(S): Ectopic endometrial stromal cells (ESCs) were isolated and cultured with IL-4. Ectopic endometriotic tissues were immunostained for eotaxin and IL-4. MAIN OUTCOME MEASURE(S): Gene expression of eotaxin was determined by standard and real-time reverse-transcriptase polymerase chain reaction. Secretion of eotaxin from ESC was measured using specific ELISA. The immunostained sections were examined. RESULT(S): Interleukin-4 (IL-4) increased mRNA expression and protein secretion of eotaxin from ESC in a dose-dependent manner. Immunohistochemical analysis showed that eotaxin-positive cells colocalized with IL-4-positive cells and accumulated around the blood vessels in the stroma of endometriotic tissue. CONCLUSION(S): IL-4 induces eotaxin in ESCs, which might promote angiogenesis and the subsequent development of endometriosis.

Interleukin-4 stimulates proliferation of endometriotic stromal cells.

Several lines of evidence indicate that the Th2 immune response is associated with endometriosis. Although an increased concentration of interleukin (IL)-4, a typical Th2 cytokine, has been reported in endometriotic tissues, the implication of this for endometriosis has not been determined. To investigate a possible role of IL-4 in the development of endometriosis, we examined the presence of IL-4-producing cells in endometriotic tissues and the effect of IL-4 on proliferation of endometriotic stromal cells. Endometriotic stromal cells were isolated from endometriotic tissues obtained from women undergoing surgery for endometrioma. Immunohistochemistry of endometriotic tissues revealed that IL-4-positive cells were abundant in the stroma. The effect of IL-4 on proliferation of endometriotic stromal cells was studied using cell counting and BrdU incorporation assays. IL-4 (0.1 to 10 ng/ml) significantly increased cell number and BrdU incorporation in a dose-dependent manner, and the proliferative effect of IL-4 was inhibited by anti-IL-4 receptor antibody. IL-4-induced activation of mitogen-activated protein kinases in endometriotic stromal cells was examined by Western blotting. IL-4 induced phosphorylation of p38 mitogen-activated protein kinase, stress-activated protein kinase/c-Jun kinase, and p42/44 mitogen-activated protein kinase and inhibitors of these kinases suppressed IL-4-induced proliferation of endometriotic stromal cells. These findings suggest that proliferation of endometriotic stromal cells induced by locally produced IL-4 is involved in the development of endometriosis.