HSF1通过上调MTDH促进子宫内膜异位症在位内膜间质细胞的上皮-间质转化  

Effect of HSF1 on eutopic endometrial stromal cells in endometriosis by up regulating MTDH

作　　者：李娜[1] 李瑞娜 李娇[1] 赵静妮[1] 朱敏玲 付晶[1] LI Na;LI Ruina;LI Jiao;ZHAO Jingni;ZHU Minling;FU Jing(Department of Obstetrics and Gynecology,the First Affiliated Hospital of Xi’an Jiaotong University,Xi'an,Shaanxi 710061,China)

机构地区：[1]西安交通大学第一附属医院妇产科,陕西西安710061

出　　处：《中国优生与遗传杂志》2022年第2期177-184,共8页Chinese Journal of Birth Health & Heredity

基　　金：陕西省重点研发计划项目(2020SF-041)。

摘　　要：目的 探讨热休克因子-1(HSF1)和星形胶质细胞升高基因-1(MTDH)对子宫内膜异位症在位内膜间质细胞迁移、侵袭和上皮-间质转化(epithelial-mesenchymal transition,EMT)的作用。方法 分别获取子宫内膜异位症患者(n=20)的在位和异位子宫内膜组织,采用RT-qPCR法、Western blot法和免疫组织化学染色法分别检测各在位和异位子宫内膜组织中HSF1和MTDH的表达情况;离体原代培养在位内膜间质细胞(EuSCs),并通过转染过表达HSF1,采用RT-qPCR法和Westernblot法对过表达HSF1效率进行验证;采用体外侵袭实验和划痕实验对EuSCs组,EuSCs转染pcDNA3.1组和EuSCs转染pcDNA3.1-HSF1组细胞的侵袭和迁移能力变化进行检测;采用细胞形态学成像和westernblot法分别对上述3组细胞的EMT表型和标志物,包括E-钙黏素、N-钙黏素、蜗牛蛋白和波形蛋白的表达水平变化进行检测。结果 与在位子宫内膜组织中HSF1和MTDH的表达水平相比,来源于子宫内膜异位症患者(n=20)的异位子宫内膜组织中对应蛋白质的表达水平升高,差异具有统计学意义(P<0.01);RT-qPCR法和Westernblot法证实在位内膜间质原代细胞中成功过表达HSF1;过表达HSF1后的在位内膜间质原代细胞,相较于过表达HSF1之前,MTDH表达被上调(P<0.01);细胞迁移(P<0.01)和侵袭能力增强(P<0.01),差异具有统计学意义;细胞形态更趋于成纤维样细胞表型,EMT标志物N-钙黏素、蜗牛蛋白和波形蛋白表达被上调(P<0.01),E-钙黏素的表达被抑制(P<0.05),差异具有统计学意义。结论 HSF1通过上调MTDH,调节子宫内膜异位症在位内膜间质细胞的EMT、侵袭和转移能力。Objective To explore the roles of heat shock factor 1(HSF1) and astrocyte elevated gene-1(MTDH) on the eutopic endometrial stromal cells migration, invasion ability and EMT. Methods Obtain the eutopic and ectopic endometrial tissues of patients with endometriosis(n=20). RT-qPCR, western blot and immunohistochemical staining were used to detect the expression of HSF1 and MTDH in eutopic and ectopic endometrial tissues, respectively. Primary endometrium mesenchymal cells(EuSCs) were cultured and overexpressed HSF1 by transfection. The efficiency of overexpression of HSF1 was verified by RT-qPCR and western blot. In vitro invasion assay and wound closure assay were used to detect the invasion and migration ability of EuSCs group, EuSCs transfected with pcDNA3.1 group and EuSCs transfected with pcDNA3.1-HSF1 group cells. Cell morphological imaging was used to detect the EMT phenotypes of the above three groups of cells. Western blot was used to detect the EMT markers of the above three groups of cells, including E-cadherin, N-cadherin, snail and vimentin expression. Results The expression levels of HSF1 and MTDH in ectopic endometrial tissues from patients with endometriosis(n=20) were significantly higher than in eutopic endometrial tissues(P<0.01). RT-qPCR and western blot results showed that HSF1 was over-expressed in primary endometrium mesenchymal cells(EuSCs). Compared with EuSCs, the expression of MTDH in over-expressed HSF1 EuSCs was up-regulated(P<0.01). Over-expressed HSF1 EuSCs cell migration ability(P<0.01) and invasion ability(P<0.01) was up-regulated, compared with EuSCs. Over-expressed HSF1 EuSCs cell morphology tends to be fibroblastic phenotype, the expression of EMT markers, including N-cadherin, snail and vimentin were significantly up-regulated(P<0.01), and the expression of E-cadherin was significantly down-regulated(P<0.01), compared with EuSCs. Conclusion HSF1 promotes eutopic endometrial stromal cells EMT, invasion and metastasis ability by up-regulating MTDH.

关 键 词：子宫内膜异位症 在位内膜间质细胞 热休克因子-1 星形胶质细胞升高基因-1 上皮-间质转化 

分 类 号：R711.71[医药卫生—妇产科学]