沉默干细胞转录因子Sox2对宫颈癌细胞增殖能力影响的研究  

Effect of silencing stem cell transcription factor Sox2 on proliferation of cervical cancer cells

作　　者：冀静[1] 左萍 黄康榕 刘海娟[2] 方静[1] 王月玲[1] 

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 [2]西安市第四医院妇产科,陕西西安710004

出　　处：《中国妇幼健康研究》2017年第12期1593-1596,共4页Chinese Journal of Woman and Child Health Research

基　　金：国家科学青年基金资助项目(81001162);陕西省自然科学基金重点研发计划资助项目(2017SF-015);西安交通大学第一附属医院院基金资助项目(YK201503)

摘　　要：目的探讨沉默干细胞转录因子Sox2对宫颈癌细胞增殖能力的影响及其机制。方法将干扰质粒(shRNA-Sox2)转染宫颈癌C33A细胞系,利用嘌呤霉素筛选出稳定沉默Sox2表达的C33A细胞克隆株(shRNA-Sox2-C33A)。MTT、克隆形成实验检测沉默Sox2表达对宫颈癌细胞增殖能力的影响;流式细胞仪检测细胞周期的变化;Western blot检测细胞周期素CyclinD1的表达变化。结果 shRNA-Sox2转染C33A细胞后,细胞中Sox2 mRNA表达下调(t=7.94,P<0.01)。干扰组细胞(shRNA-Sox2-C33A)增殖速度低(t=10.93,P<0.05),体外克隆形成率显著下降(t=8.30,P<0.01);细胞周期中G0/G1期细胞比例增多,S期细胞比例减少(t值分别为-4.60、4.67,均P<0.05);Western blot检测CyclinD1表达下调(t=113.79,P<0.05)。结论沉默Sox2基因表达可能通过下调CyclinD1来阻滞细胞从G1期细胞进入S期,抑制宫颈癌细胞增殖。Objective To study the influence of silencing stem cell transcription factor Sox2 on cervical cancer cell proliferation and its mechanism.Methods Interference plasmid（shRNA-Sox2）was stably transfected into cervical cancer cell line C33 A,then C33 A clonal cell line with stably silenced Sox2-expression（shRNA-Sox2-C33 A）was screened by puromycin selection.MTT assay and colony-forming experiment were used to detect effect of silencing Sox2 expression on cervical cancer cell proliferation ability.Change of cell cycle was studied by flow cytometry（FCM）,while expression of cell cycle related protein CyclinD1 was detected by Western blot.Results The mRNA expression of Sox2 was down-regulated in C33 Acells after shRNA-Sox2 transfection（t=7.94,P〈0.01）.Proliferation rate of shRNA-Sox2-C33 Agroup was low（t=10.93,P〈0.05）.Down regulation of Sox2 expression also led to decreased cloning formation ability of cervical cancer cells in vitro（t=8.30,P〈0.01）.Proportion of cells in G0/G1 phase was increased,while proportion of cells in S phase was reduced（t value was-4.60 and 4.67,respectively,both P〈0.05）.Decreased expression of CyclinD1 was detected by Western blot（t=113.79,P〈0.05）.Conclusion Silencing of Sox2 expression might stop cell from entering into S phase from G1 phase through down regulation of CyclinD1,inhibiting proliferation of cervical cancer cells.

关 键 词：Sox2基因 宫颈癌 基因沉默 细胞增殖 

分 类 号：R737.33[医药卫生—肿瘤]