小干扰RNA下调埃兹蛋白表达对子宫内膜癌HEC-1B细胞凋亡及增殖的影响  

Effect of apoptosis and proliferation of endometrial cancer HEC-1B cell by siRNA down-regulating expression of Ezrin

作　　者：畅华[1] 张雪[1] 李白雪 郭科军[1] 姚霁航[1] Chang Hua;Zhang Xue;Li Baixue;Guo Kejun;Yao Jihang(Department of Gynecology, the First Affiliated Hospital of China Medical University, Shenyang 110001, China)

机构地区：[1]中国医科大学附属第一医院妇科,沈阳110004

出　　处：《中国综合临床》2019年第3期255-259,共5页Clinical Medicine of China

摘　　要：目的观察埃兹蛋白(Ezrin)在子宫内膜癌细胞中表达及下调后子宫内膜癌细胞周期、凋亡及增殖变化,探讨其是否可能作为靶向治疗候选基因。方法对来源于2017年2月中国医学科学院上海细胞研究所子宫内膜癌细胞株根据干预方式分为siEzrin转染组及空白对照组,采用蛋白免疫印迹(Western blot)及实时定量多聚酶链反应(quantificational real-time polymerase chain reaction,qRT-PCR)检测Ezrin及mRNA,采用小干扰RNA(small interfering RNA,siRNA)转染HEC-1B细胞,对Ezrin表达进行下调。流式细胞术检测细胞周期及凋亡,四甲基偶氮唑盐微量酶反应比色法[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]检测细胞增殖。结果Western blot显示,Ezrin蛋白在子宫内膜癌细胞株Ishikawa(31.742±5.832)、HEC-1A(16.326±3.135)、HEC-1B(17.636±4.426)及KLE(14.862±5.109)细胞株中均有表达;qRT-PCR显示,mRNA在子宫内膜癌细胞株Ishikawa(2.513±0.725)、HEC-1A(1.655±0.692)、HEC-1B(3.237±0.411)及KLE(0.962±0.235)细胞株中均有表达,在HEC-1B细胞中Ezrin蛋白及mRNA表达最高(F=6.173,P<0.05;F=7.042,P<0.05);流式细胞术显示,与空白对照组比较,siEzrin转染组HEC-1B细胞停留在G1期及G2期比例少,S期比例增高(t=3.118,P<0.05;t=5.435,P<0.05;t=3.332,P<0.05),Ezrin下调后HEC-1B细胞凋亡率从(9.84±2.37)%增加至(17.64±5.96)%(t=8.963,P<0.01);MTT检测显示,与空白对照组比较,在72 h及96 h siEzrin转染组HEC-1B细胞增殖能力降低(t=3.209,P<0.05;t=3.726,P<0.05)。结论下调Ezrin表达使子宫内膜癌细胞更多停留在S期,促进其凋亡,抑制增殖,Ezrin可作为子宫内膜癌靶向治疗候选基因。Objective To observe the changes of cell cycle, apoptosis and proliferation of endometrial cancer cells after the expression and down-regulation of Ezrin in endometrial cancer cells and to explore whether Ezrin may be a candidate gene for targeted therapy. Methods Endometrial cancer cells were from Shanghai Institute of Cell Research, of Chinese Academy of Medical Sciences in February 2017 and divided into blank control group and siEzrin group according to the intervention methods.Western blot and qRT-PCR was used to detect the expression of Ezrin protein and mRNA in endometrial cell lines.Small interfering RNA(siRNA) was used to transfect HEC-1B cell and down-regulate Ezrin.Cell cycle and apoptosis were detected by flow cytometry.MTT assay was used to detect multiplication. Results Western blot showed that Ezrin protein was expressed in Ishikawa(31.742±5.832)、HEC-1A(16.326±3.135)、HEC-1B(17.636±4.426) and KLE(14.862±5.109) and qRT-PCR showed that mRNA was expressed in Ishikawa (2.513±0.725), HEC-1A (1.655±0.692), HEC-1B (3.237±0.411) and KLE (0.962±0.235) cell lines, and expressed highest in HEC-1B cells (F=6.173, P<0.05;F=7.042, P<0.05). Flow cytometry assay showed that compared with blank control group less cells stayed in G1 phase and G2 phase, more stayed in S phase (t=3.118, P<0.05;t=5.435, P<0.05;t=3.332, P<0.05). The apoptotic rate of HEC-1B cells increased from (9.84±2.37)% to (17.64±5.96)%(t=8.963, P<0.01) after Ezrin was down-regulated.MTT assay showed that the proliferation of HEC-1B cells in 72 h and 96 h siEzrin transfection group was lower than that in blank control group (t=3.209, P<0.05;t=3.726, P<0.05). Conclusion Down-regulating of Ezrin may promote more endometrial cancer cells stay in S phase and promote apoptosis, inhibit proliferation, Ezrin may become target candidate gene in target therapy.

关 键 词：子宫内膜癌 埃兹蛋白 细胞周期 凋亡 增殖 

分 类 号：R737.33[医药卫生—肿瘤]