稳定干扰Pin1基因的食管癌细胞株的筛选及其生物学特征  

Screening and characterization of stable Pin1-knockdown esophageal squamous cell carcinoma cell strains

作　　者：李玲[1] 陈平[1] 连鸿凯[2] 赵继敏[1] 马俊芬[1] 董子明[1] 

机构地区：[1]郑州大学基础医学院病理生理教研室,河南省郑州市450001 [2]郑州市骨科医院,河南省郑州市450001

出　　处：《世界华人消化杂志》2010年第14期1485-1488,共4页World Chinese Journal of Digestology

摘　　要：目的:筛选稳定干扰Pin1的食管癌细胞系,研究Pin1表达与食管癌细胞生物学特征的关系.方法:将针对Pin1基因的shRNA(pmU6-Pin1)转染入EC1细胞,经G418加压筛选稳定表达Pin1小干扰RNA的细胞株,用免疫印迹法检测细胞中Pin1的表达,确定筛选细胞系的正确性.通过MTT实验以及流式细胞仪检测Pin1抑制对食管癌细胞增殖、凋亡的影响.结果:Western blot检测结果显示,Pin1蛋白表达被成功抑制,建立了稳定表达Pin1小干扰RNA的细胞株(shPin1).与正常EC1细胞比较,MTT实验和流式细胞术实验结果表明基因沉默Pin1抑制食管癌细胞增殖(抑制率为51.8%),诱导细胞凋亡(凋亡率为46.39%);并增加了食管癌细胞EC1对顺铂的敏感性,抑制食管癌细胞增殖以及诱导细胞凋亡的能力均有所增强,抑制率从23.5%上升到61.0%,凋亡率从26.10%上升到58.95%.结论:稳定干扰Pin1食管癌细胞系的建立为进一步研究Pin1在食管癌中的作用提供了研究平台;基因沉默Pin1增加了食管癌细胞对顺铂的敏感性.AIM：To screen stable peptidyl-prolyl cis/trans isomerase（Pin1）-knockdown esophageal squamous cell carcinoma cell strains and to study the effects of Pin1 down-regulation on the biological characteristics of esophageal squamous cell carcinoma cells（EC1）.METHODS：After EC1 cells were transfected with pmU6-Pin1 plasmid,cell strains stably expressing Pin1 siRNA were selected in the presence of G418.The expression of Pin1 protein was detected by Western blotting.After stable Pin1-knockdown cell strains were obtained,the impact of Pin1 knockdown on cell growth and apoptosis was detected by MTT assay and flow cytometry,respectively.RESULTS：Western blot analysis showed that the expression of Pin1 protein was effectively inhibited in EC1 cell strains stably expressing Pin1 siRNA.MTT assay and flow cytometry showed that Pin1 knockdown inhibited cell proliferation（the reduced rate of cell proliferation：51.8%） and induced cell apoptosis（apoptosis rate：46.39%）.Inhibition of Pin1 could significantly increase the sensitivity of EC1 to cisplatin（CDDP） since the reduced rate of cell proliferation increased from 23.5% to 61.0% and the apoptosis rate from 26.10% to 58.95%.CONCLUSION：Stable Pin1-knockdown esophageal squamous cell carcinoma cell strains provide a basis for studying Pin1 function in esophageal squamous cell carcinoma.The observation that Pin1 knockdown could increase the sensitivity of EC1 cells to CDDP provides new insight into the therapy of esophageal squamous cell carcinoma.

关 键 词：稳定干扰 PIN1 增殖 凋亡 顺铂 

分 类 号：R735.1[医药卫生—肿瘤]