干扰CIP2A表达对卵巢癌细胞增殖和凋亡影响研究  

Influence of CIP2A to cell proliferation and apoptosis of human ovarian cancer cells

作　　者：方媛媛[1] 李一宁[1] 王秀霞[1] 张淑兰[1] 

机构地区：[1]中国医科大学附属盛京医院妇产科,辽宁沈阳110004

出　　处：《中华肿瘤防治杂志》2014年第6期424-427,432,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：辽宁省教育厅高等学校科研项目(LT2010106)

摘　　要：目的:探讨干扰蛋白磷酸酶2A癌性抑制因子(cancerous inhibitor of protein phosphatase 2A,CIP2A)表达对卵巢癌细胞增殖和凋亡的影响。方法:应用siRNA干扰卵巢癌细胞A2780和SKOV3的CIP2A表达后,检测细胞增殖、凋亡和相关基因表达水平。MTT和集落形成试验检测细胞增殖水平;流式细胞术检测细胞周期和凋亡水平;实时定量PCR和蛋白质印迹法检测CIP2A、Cyclin D1、c-myc、p-Rb、Bcl-2和p-AKT等细胞周期相关蛋白的表达水平。结果:siRNA敲减A2780和SKOV3细胞的CIP2A表达,敲减效率A2780为81%,SKOV3为78%。干扰后细胞增殖受抑,集落形成数量降低,A2780细胞,对照组为349±19,CIP2AsiRNA为207±12,P=0.001;SKOV3细胞,对照组为288±16,CIP2AsiRNA为106±8,P=0.001。干扰后细胞周期受阻碍,G1期细胞比例增加,A2780细胞,对照为45.5±2.8,CIP2AsiRNA为55.8±3.1,P=0.045;SKOV3细胞,对照为55.2±2.3,CIP2AsiRNA为65.6±3.2,P=0.041。干扰后S期细胞比例下降,A2780细胞,对照为38.6±1.9,CIP2AsiRNA为31.3±2.1,P=0.044;SKOV3细胞,对照为29.5±1.7,CIP2AsiRNA为22.3±1.5,P=0.041。干扰CIP2A表达可促进紫杉醇诱导的细胞凋亡,CIP2AsiRNA凋亡率,A2780细胞为23.63%,SKOV3细胞为20.69%;对照组凋亡率A2780细胞为14.25%,SKOV3细胞为12.75%。蛋白质印迹法结果显示,干扰CIP2A表达可下调Cyclin D1、c-myc、p-Rb、Bcl-2和p-AKT的表达。结论:CIP2A对卵巢癌细胞增殖和凋亡起重要作用,可能成为卵巢癌治疗的新靶点。OBJECTIVE： To investigate the influence of CIP2A siRNA to ovarian cancers cell proliferation and apop- tosis. METHODS： We performed siRNA knockdown in A2780 and SKOV3 ovarian cell lines and examine cell prolifera- tion,apoptosis and related gene expression. MTT and colony formation assay were carried out to assess cell proliferation; Flow cytometry was carried out to assess cell cycle and apoptosis level;Real time PCR and Western blot were carried out to assess the expression of CIP2A and cell cycle related proteins, Cyclin D1, c-myc, p-Rb, Bcl-2 and p-AKT. RESULTS： CIP2A knockdown in ovarian cancer cell lines（knockdown rate of A2780 was 81 %, while that of SKOV3 was 78% ） in- hibited proliferation,decreased colony number （A2780, control group was 349±19, CIP2A siRNA group was 207 ±12, P = 0.001 ; SKOV3, control group was 288 ± 16, CIP2A siRNA group was 106 ± 8, P = 0.001）, blocked cell cycle progres- sion,increased the percentage of G1 stage cells （A2780, control group was 45.5 ± 2.8, CIP2A siRNA group was 55.8 ± 3.1, P = 0. 045 ; SKOV3, control group was 55.2 ±2.3, CIP2A siRNA group was 65.6 ± 3.2, P = 0. 041 ）, and decreased the percentage of S stage cells （A2780, control group was 38.6 ± 1.9, CIP2A siRNA group was 31.3±2.1, P = 0. 044; SKOV3, control group was 29.5 ± 1.7, CIP2A siRNA group was 22.3 ±1.5, P= 0. 041）, and increased paclitaxel induced apoptosis（CIP2A siRNA group, A2780 was 23.63 %, SKOV3 was 20.69 % ; Control group, A2780 was 14.25%, SKOV3 was 12.75 % ）. Futhermore, CIP2A knockdown downregulated Cyclin D1, c-myc, phospho-Rb, Bcl-2 and phospho-AKT ex- pression. CONCLUSION： These results validated CIP2A plays an important role to ovarian cancer cells proliferation and apoptosis,and CIP2A is a promising therapeutic target of ovarian cancers.

关 键 词：卵巢肿瘤 CIP2A 增殖 凋亡 干扰RNA 

分 类 号：R737.31[医药卫生—肿瘤]