Department of Obstetrics and Gynecology, College of Medicine, Catholic University of Korea, 505, Banpo-dong, Seocho-gu, Seoul 137-701, Korea.
J Clin Endocrinol Metab. 2012 Sep;97(9):3224-30. doi: 10.1210/jc.2012-1538. Epub 2012 Jul 3.
Müllerian inhibiting substance (MIS) is produced in Sertoli cells of fetal testis and causes regression of müllerian ducts in male embryos. MIS also can induce the cell cycle arrest and apoptosis in müllerian duct-derived tumors in vivo and in vitro.
Our objective was to investigate the expression of MIS type II receptor (MISR II) and whether MIS can inhibit the proliferation and induce apoptosis in primary cultures of endometrial stromal cells (ESC) of endometriosis.
In vitro experiments were performed in the university research laboratory.
Tissue samples from 12 patients who had undergone evisceration for ovarian endometrial cysts were included in this study.
The expression of MISR II in ESC was investigated by immunohistochemistry. The cell viability and apoptosis in ESC treated with MIS was measured by methylthiazoletetrazolium assay and annexin V analysis. The expression of regulatory proteins in ESC treated with MIS was shown by Western blotting.
ESC showed specific immunostaining for the MISR II. ESC treated with MIS exhibited 32% growth inhibition (P = 0.0001). The changes in cell cycle distribution after MIS exposure at 72 h demonstrated that S and G(2)M phases were decreased; G(0)G(1) and sub-G(0)G(1) phases were increased. ESC treated with MIS showed 13.72% annexin V-fluorescein isothiocyanate positivity. In the ESCs, which contain defective p16, MIS increased the expression of pocket proteins p107 and p130 and decreased E2F transcription factor 1.
The results support a central role for MIS in endometriosis. Although the precise mechanism of MIS-mediated inhibition of ESC growth has not been fully defined, these data suggest that MIS has activity against ESC in vitro and may also be an effective targeted therapy for endometriosis.
苗勒管抑制物质（MIS）由胎儿睾丸的支持细胞产生，可导致男性胚胎中苗勒管退化。MIS 还能诱导体内和体外苗勒管源性肿瘤的细胞周期停滞和凋亡。
本研究旨在探讨 MIS Ⅱ型受体（MISR II）的表达情况，以及 MIS 是否能抑制子宫内膜异位症患者原代子宫内膜基质细胞（ESC）的增殖并诱导其凋亡。
在大学研究实验室进行体外实验。
本研究纳入了 12 例行卵巢子宫内膜囊肿剔除术患者的组织样本。
通过免疫组织化学法检测 ESC 中 MISR II 的表达情况。通过噻唑蓝比色法和 Annexin V 分析检测 MIS 处理后的 ESC 细胞活力和凋亡情况。通过 Western 印迹法显示 MIS 处理后的 ESC 中调节蛋白的表达情况。
ESC 显示出特异性的 MISR II 免疫染色。MIS 处理后的 ESC 生长抑制率为 32%（P = 0.0001）。MIS 暴露 72 h 后细胞周期分布的变化表明，S 期和 G2/M 期减少，G0/G1 期和 sub-G0/G1 期增加。MIS 处理后的 ESC 中 Annexin V-异硫氰酸荧光素阳性率为 13.72%。在含有缺陷型 p16 的 ESC 中，MIS 增加了 pocket 蛋白 p107 和 p130 的表达，降低了 E2F 转录因子 1 的表达。
这些结果支持 MIS 在子宫内膜异位症中的核心作用。尽管 MIS 介导的 ESC 生长抑制的确切机制尚未完全确定，但这些数据表明 MIS 在体外对 ESC 具有活性，也可能是子宫内膜异位症的有效靶向治疗方法。
