CAPN7促进人子宫内膜间质细胞增殖的初步研究  

The role of CAPN7 in human endometrial stromal cells proliferation

作　　者：刘红玉[1] 朱丽华[1] 沈晓月[1] 金晓艳[1] 颜桂军[2] 孙海翔[1,2] 

机构地区：[1]南京医科大学鼓楼临床医学院,江苏南京210029 [2]南京大学医学院附属鼓楼医院生殖医学中心,江苏南京210008

出　　处：《中国优生与遗传杂志》2013年第2期27-31,38,共6页Chinese Journal of Birth Health & Heredity

基　　金：国家自然科学基金(No.81170570;No.30900727;No.81070492);"六大人才高峰"项目基金(No.2010-WS-058);南京市卫生局资金(QYK09172);留学回国人员科研启动基金(No.2011-1139)资助项目

摘　　要：目的子宫内膜异位症是引起育龄妇女不孕的主要疾病之一,研究表明子宫内膜异位症患者在位子宫内膜间质细胞增殖增强,但具体机制不清。我们研究发现半胱氨酸蛋白酶CAPN 7在子宫内膜异位症患者在位子宫内膜组织中表达显著增高,本研究旨在探讨CAPN 7对子宫内膜间质细胞增殖的影响。方法提取正常育龄妇女及子宫内膜异位症患者在位子宫内膜组织总蛋白,通过Western Blotting检测CAPN 7蛋白表达。制备Ad-GFP-CAPN 7重组腺病毒,采用CCK-8增殖试剂盒和Western Blotting结合腺病毒介导的CAPN 7基因过表达及小干扰RNA介导的CAPN 7基因沉默实验,研究CAPN 7对人子宫内膜间质细胞增殖的影响。结果 CAPN 7蛋白在子宫内膜异位症患者在位子宫内膜组织中的表达明显高于正常育龄妇女子宫内膜组织;成功获得滴度为3.5×1010ifu/mL的重组GFP-CAPN 7腺病毒,人子宫内膜间质细胞感染该腺病毒后,可表达带有绿色荧光蛋白的CAPN 7融合蛋白;伴随CAPN 7蛋白表达量的增加子宫内膜间质细胞中CyclinD3蛋白的表达增加,同时显著增加细胞的增殖能力;基因沉默子宫内膜间质细胞中内源性CAPN 7表达后,子宫内膜间质细胞中CyclinD3蛋白的表达降低且增殖能力降低15%。结论 CAPN 7通过调控细胞周期蛋白CyclinD3的表达促进人子宫内膜间质细胞的增殖。Objective： Endometriosis is one of the leading causes of female infertility,many reports have shown that the proliferation of eutopic endometrial stromal cells from women with a diagnosis of endometriosis is higher than that from normal fertile women,but the pathogenesis is still unclear.Our result suggested that calcium-activated neutral protease 7（CAPN 7） is highly expressed in eutopic endometrium from endometriosis.This study aimed to explore the role of CAPN 7 in human endometrial stromal cells（hESCs） proliferation.Methods： Western Blotting was used to detect the expression of CAPN 7 in eutopic endometrium from endometriosis patients and normal fertile women.We constructed adenovirus-GFP-CAPN 7 to infect hESCs.Through overexpression of CAPN 7 by infection with Ad-GFP-CAPN 7 and knockdown the endogenous CAPN 7 expression by transfection with siCAPN 7,we studied the effects of CAPN 7 on hESCs proliferation using Cell Counting Kit-8（CCK-8） and Western Blotting.Results： CAPN 7 was up-regulated in endometriosis.We successfully obtained a total of 3.5×1010 ifu/mL adenovirus-GFP-CAPN 7,which could highly express the GFP-CAPN 7 fusion protein in hESCs.When CAPN 7 was overexpressed,the protein expression of CyclinD3 increased and the proliferation of hESCs increased by 1.5-fold.But when CAPN 7 was knocked down,CyclinD3 expression markedly decreased and the proliferation reduced by 15%.Conclusion： CAPN 7 promoted the proliferation of hESCs and may contribute to a high expression of CyclinD3.

关 键 词：CAPN7 人子宫内膜间质细胞 增殖 

分 类 号：R711.71[医药卫生—妇产科学]