血管内皮生长因子调节蛋白激酶C表达对子宫内膜异位症间质细胞侵袭性的影响  

Effects of Vascular Endothelial Growth Factor(VEGF)Regulating Protein Kinase C Expression on the Invasiveness of Stromal Cells in Endometriosis

作　　者：王芳 王丽春 韩彩玲[3] 马少寒 陈华[3] 樊杨[1] WANG Fang;WANG Lichun;HAN Cailing;MA Shaohan;CHEN Hua;FAN Yang(Department of Obstetrics and Gynecology,People’s Hospital of Ningxia Hui Autonomous Region,Third Clinical Medical College of Ningxia Medical University,Yinchuan 750002,China;Maternal and Child Health Hospital,Yinchuan 750004,China;Department of Gynecology,General Hospital of Ningxia Medical University,First Clinical Medical College of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏回族自治区人民医院妇产科,宁夏医科大学第三临床医学院,银川750002 [2]银川市妇幼保健院产科,银川750004 [3]宁夏医科大学总医院妇科,宁夏医科大学第一临床医学院,银川750004

出　　处：《宁夏医科大学学报》2024年第8期757-764,共8页Journal of Ningxia Medical University

基　　金：宁夏自然科学基金项目(2023AAC03469);宁夏医科大学2020年大学生创业创新计划项目。

摘　　要：目的探讨血管内皮生长因子(VEGF)调节蛋白激酶C(PKC)表达对子宫内膜异位症(EMs)间质细胞侵袭性的影响。方法收集因卵巢EMs行手术并经术后病理证实为EMs的20例患者在位及异位内膜组织,分为EMs在位内膜组及EMs异位内膜组,收集同期因其他卵巢良性肿瘤行手术的10例患者子宫内膜作为对照组,应用免疫组织化学染色法及Western blot法检测各组子宫内膜中VEGF及PKC蛋白表达情况;分离培养EMs在位内膜间质细胞与非EMs子宫内膜间质细胞,应用细胞免疫鉴定法鉴定间质细胞,应用免疫荧光染色法及Western blot法检测各组间质细胞中VEGF和PKC蛋白表达及定位情况;采用VEGF-siRNA转染EMs子宫内膜间质细胞,采用Western blot法检测沉默VEGF后EMs间质细胞的VEGF、PKC蛋白表达变化,采用Transwell法检测其侵袭性的变化。结果与对照组相比,EMs在位内膜组及EMs异位内膜组VEGF、PKC蛋白表达均升高,且EMs异位内膜组中的表达水平高于EMs在位内膜组(P均<0.01)。EMs子宫内膜间质细胞中VEGF蛋白、PKC蛋白荧光染色均能看到绿色荧光,VEGF蛋白在子宫内膜间质细胞质和细胞核中均有表达,而PKC蛋白主要表达在子宫内膜间质细胞质中。Western blot结果显示,与对照组相比,EMs组VEGF及PKC蛋白表达均升高(P均<0.001);培养72 h后siRNA转染效率最高;与si R-NC组比较,VEGF-siRNA转染后EMs间质细胞中VEGF、PKC蛋白表达水平均降低(P均<0.001);Transwell结果显示,与siR-NC组及空白对照组比较,VEGF-siRNA转染后,EMs间质细胞的侵袭细胞数减少(P均<0.001)。结论VEGF可能通过调节PKC蛋白表达,继而改变细胞侵袭活性,诱导EMs的发生和发展。Objective To investigate the effects of vascular endothelial growth factor(VEGF)regulating protein kinase C(PKC)expression on the invasiveness of stromal cells in endometriosis(EMs).Methods Twenty patients with ovarian(EMs)and EMs confirmed by intraoperative and postoperative pathology were collected.The eutopic and ectopic endometrial tissues were collected and divided into EMs eutopic group and EMs ectopic group.The eutopic and ectopic endometrial tissues of 20 patients operated for ovarian endometriosis and pathologically confirmed as EMs after postoperative surgery were collected and divided into EMs.20 patients operated for other benign ovarian tumors in the same period were collected as the control group.Immunofluorescence staining was used to detect the expression and localization of VEGF and PKC proteins in the stromal cells of the EMs group.Western blot was used to detect the expression of VEGF and PKC protein in the control group and EMs group.EMs stromal cells were transfected by VEGF-siRNA gene silencing method,and the changes of VEGF and PKC protein expression after intervention were detected by Western blot.Transwell method was used to detect the invasiveness of cells after intervention.Results Compared with the control group,the expression of VEGF and PKC proteins in the EMs eutopic and ectopic groups increased,and the expression levels in the ectopic endometrium group were higher than those in the eutopic endometrium group(P all<0.01).The results of cell immunofluorescence staining showed that green fluorescence was produced in both VEGF protein and PKC protein in the endometrial stromal cells of EMs.VEGF protein was expressed in both the cytoplasm and nucleus of the endometrial stromal cells,while PKC protein was mainly expressed in the cytoplasm of the endometrial stromal cells.Western blot results showed that compared with the control group,the expressions of VEGF and PKC proteins in the EMs group were increased(P all<0.001).The results showed that the siRNA transfection efficiency was the high

关 键 词：子宫内膜异位症 血管内皮生长因子 蛋白激酶C 细胞侵袭性 

分 类 号：R711.71[医药卫生—妇产科学]