基于ROCK-2/Moesin信号通路探究下调SIP1对子宫内膜癌细胞凋亡侵袭迁移的影响  

Effects of Downregulation of SIP1 on Apoptosis Invasion and Migration of Endometrial Cancer Cells Based on ROCK-2/Moesin Signaling Pathway

作　　者：刘雪洁 李晓妍 张凤 LIU Xuejie;LI Xiaoyan(Baoding Second Central Hospital,Hebei Baoding 072750,China)

机构地区：[1]河北省保定市第二中心医院妇科,河北保定072750 [2]河北省第八人民医院外一科,河北石家庄050000

出　　处：《河北医学》2023年第3期364-369,共6页Hebei Medicine

基　　金：河北省医学科学研究重点课题计划,(编号:20200264)。

摘　　要：目的:探究下调SIP1对子宫内膜癌细胞凋亡、迁移、侵袭影响及其作用机制。方法:人子宫内膜癌细胞HEC-1A细胞分为对照组,NC组,SIP1下调组(转染si-SIP1),Y27632组(ROCK抑制剂),SIP1下调组+Y27632组(转染si-SIP1+ROCK抑制剂),通过流式细胞技术、Transwell实验分别检测各组细胞凋亡、迁移能力和侵袭能力,通过Western blot检测各组细胞凋亡关键蛋白B淋巴细胞瘤-2基因(Bcl-2)、细胞凋亡基因Bcl-2相关蛋白x(Bax)、Caspase家族蛋白3(Caspase3)、Rho相关激酶2(ROCK-2)和Moesin蛋白表达水平。20只SPF级BALB/c小鼠通过皮下荷瘤建立子宫内膜癌荷瘤模型,对照组(n=10)采用人子宫内膜癌HEC-1A细胞荷瘤,SIP1下调组(n=10)小鼠采用下调SIP1的人子宫内膜癌HEC-1A细胞荷瘤,每3天用游标卡尺测量瘤体积,连续测量21d绘制肿瘤生长曲线,实验终点测量每只小鼠肿瘤质量并检测肿瘤组织中ROCK-2及Moesin蛋白表达水平。结果:细胞实验结果表明,与对照组和NC组相比,SIP1下调组、Y27632组、SIP1下调+Y27632组细胞凋亡比例显著上升,细胞迁移数量减少,细胞侵袭数量减少(P<0.01),NC组与对照组没有显著的统计学差异(P>0.05)。Western blot结果表明,与对照组和NC组相比,SIP1下调组、Y27632组、SIP1下调+Y27632组细胞ROCK和Moesin蛋白表达水平有降低趋势(P<0.01)。小鼠实验结果表明,与对照组相比,下调SIP1后小鼠肿瘤生长速度显著降低,肿瘤质量显著降低(P<0.01),肿瘤组织ROCK-2及Moesin蛋白表达水平显著降低(P<0.01)。结论:下调SIP1可导人子宫内膜癌细胞HEC-1A细胞凋亡,抑制其迁移和侵袭能力,介导ROCK-2/Moesin信号通路。Objective:To investigate the effect of down-regulation of SIP1 on apoptosis,migration and invasion of endometrial cancer cells and its mechanism.Methods:Human endometrial carcinoma cell line HEC-1A was divided into control group,NC group,SIP1 down-regulated group(transfected with si-SIP1),Y27632 group(ROCK inhibitor),SIP1 down-regulated group+Y27632 group(transfected with si-SIP1+ROCK inhibitor).The apoptosis,migration ability and invasion ability of each group were detected by flow cytometry and Transwell assay.The expression levels of apoptosis key protein b-lymphoma-2 gene(Bcl-2),apoptosis gene Bcl-2 related protein X(Bax),caspase family protein 3(caspase 3),Rho related kinase 2(ROCK-2)and Moesin were detected by Western blot.Twenty SPF-grade BALB/c mice were tumor-loaded subcutaneously to establish an endometrial cancer tumor model.The control group(n=10)was tumor-loaded with human endometrial cancer HEC-1A cells,and the SIP1 down-regulated group(n=10)was tumor-loaded with human endometrial cancer HEC-1A cells with down-regulated SIP1.Tumor volume was measured with vernier caliper every 3 days,and the tumor growth curve was drawn for 21 days.At the end point,the tumor mass of each mouse was measured and the expression levels of ROCK-2 and Moesin protein in tumor tissues were detected.Results:The results of cell experiment showed that compared with the control group and NC group,the proportion of apoptosis,the number of cell migration and cell invasion in SIP1 down regulation group,Y27632 group and SIP1 down regulation+Y27632 group increased significantly(P<0.01),there was no significant difference between NC group and control group(P>0.05).The results of Western blot showed that the expression levels of ROCK-2 and Moesin in SIP1 down regulated group,Y27632 group and SIP1 down regulated+Y27632 group decreased compared with the control group and NC group(P<0.01).The mice experiment revealed that compared with the control group,the tumor volume,tumor mass and the expression levels of ROCK-2 and Moesin in tumor t

关 键 词：SIP1 子宫内膜癌 凋亡 迁移 ROCK-2/Moesin信号通路 

分 类 号：R737.33[医药卫生—肿瘤]