siRNA干扰Chk1抑制人胃癌BGC823细胞增殖的实验研究  

Experimental study of checkpoint kinase l siRNA transfection on growth inhibition of human gastric cancer cell line BGC823

作　　者：凌晖[1] 朱亚平[1] 刘芳[1] 李艳兰[1] 文玲[1] 苏琦[1] 

机构地区：[1]南华大学肿瘤研究所,病理学教研室,湖南衡阳421001

出　　处：《中国癌症杂志》2010年第7期526-530,共5页China Oncology

基　　金：国家自然科学基金资助项目(项目编号:30600285)

摘　　要：背景与目的:细胞周期检测点激酶1(checkpoint kinase1,Chk1)与肿瘤的关系及其在肿瘤发生、发展及治疗中的作用是目前研究的热点。为探索Chk1在人类肿瘤细胞中对生长增殖活性和细胞周期的调控作用,本研究旨在观察siRNA干扰Chk1基因对人胃癌BGC823细胞生长及周期的影响。方法:采用RNAi技术抑制BGC823细胞中Chk1表达,采用Western blot检测转染前后Chk1蛋白表达情况,实验分为未转染组、脂质体组和Chk1siRNA转染组三组。然后采用MTT法和流式细胞术分析Chk1基因沉默对人胃癌BGC823细胞生长及周期的影响。结果:Western blot检测结果显示,Chk1siRNA转染24h后,BGC823细胞中Chk1蛋白相对灰度值为0.11±0.08,明显弱于未转染对照组的0.66±0.21和脂质体转染组的0.70±0.25(P<0.05),而未转染对照组和脂质体转染组之间比较,差异无统计学意义(P>0.05)。转染Chk1siRNA的BGC823细胞其Chk1蛋白表达量下降了83.1%。MTT法检测结果显示,Chk1蛋白表达缺失可导致BGC823细胞增殖活性下降,其增殖抑制率为48.1%(P<0.05)。FCM法检测结果显示,Chk1基因沉默后细胞周期在G0/G1期发生停滞,未转染组24和48h后细胞在G0/G1期为48.3%和50.2%,转染Chk1siRNA组细胞在转染24和48h后G0/G1期增加至63.9%和66.1%,增殖指数(PI)由51.7%(24h)和49.8%(48h)减少至36.2%(24h)和34.4%(48h)。结论:Chk1siRNA转染可明显抑制人胃癌细胞增殖,且其抑制增殖作用与诱导G0/G1期阻滞有关。Background and purpose：Checkpoint kinase 1 became an investigative hot spot,this hot spot involves tumorigenesis,development and therapy.In order to explore Chk1＇s regulating role in human tuomr cell growth and cycle,the effect of Chk1 siRNA transfection was investigated in terms of the cell growth and cycle in human gastric cancer cell line BGC823.Methods：The siRNA targeting at Chk1 gene was transfected into BGC823 cells.The protein expression of Chk1 was detected by Western blot.The BGC823 cells were divided into three groups：Untransfection,Lipofectamine and Chk1 siRNA-transfected group.Cell viability and cell cycle rates were determined via MTT assay and FCM.Results：The gray values of Chk1 protein was （0.11±0.08）,which was lower than in the untransfected group （0.66±0.21） as well as in the liposome transfected group （0.70±0.25） （P〈0.05）.However,there was no difference between untransfected group and liposome transfected group （P〉0.05）.The Chk1 expression at the protein level in Chk1 siRNA-transfected group was reduced by 83.1% as compared with that in the negative control group （P〈0.05）.The lack of Chk1 expression resulted in decreased cellular proliferation activity.Its inhibition rate was 48.1% （P〈0.05）.Further investigation revealed that the cell cycle was delayed at the G0/G1 phase with a remarkable arrest that occurred in the cases of Chk1 siRNA （P〈0.05）.The rate of G0/G1 phase in the untransfected group was 48.3% （24 h） and 50.2% （48 h）,whereas this number increased to 63.9% （24 h） and 66.1% （48 h） in the Chk1 siRNA-transfected group.Cell proliferating index （PI） obviously decreased from 51.7% （24 h） and 49.8% （48 h） to 36.2% （24 h） and 34.4% （48 h）,respectively.Conclusion：The proliferation of human gastric cancer BGC823 cells were inhibited by Chk1 siRNA-transfection.G0/G1 arrest was involved in the inhibiting effect when there was a lack of Chk1 expression.

关 键 词：胃肿瘤 RNA干扰 细胞周期检测点激酶1 细胞周期 

分 类 号：R73-354[医药卫生—肿瘤]