Chk1/2基因沉默对人胃癌MGC803细胞增殖及周期的影响  

Effect of Chk1/2 gene silencing on proliferation and cell cycle of human gastric cancer cell line MGC803

作　　者：凌晖[1] 文玲[1] 陈真伟[1] 李振丰[1] 夏红[1] 苏琦[1] 

机构地区：[1]南华大学肿瘤研究所,湖南衡阳421001

出　　处：《中华肿瘤防治杂志》2010年第7期481-484,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30600285);南华大学博士科研启动基金(2007XQD26)

摘　　要：目的:观察Chk1/2基因沉默对人胃癌MGC803细胞增殖及周期的影响。方法:采用RNAi技术在MGC803细胞中分别将Chk1和Chk2基因沉默,运用蛋白质印变法验证,然后采用MTT实验、流式细胞术分析Chk1和Chk2基因沉默对人胃癌MGC803细胞增殖及周期的影响。结果:蛋白质印迹法结果显示,转染靶向Chk1和Chk2siRNA24h的Chk1、Chk2蛋白相对灰度值分别为0.09±0.04和0.12±0.01,明显弱于未转染对照组(0.39±0.09)和脂质体转染组(0.38±0.17),P均<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=0.458。转染Chk1、Chk2siRNA的MGC803细胞Chk1和Chk2蛋白表达分别下降85.0%和83.4%。MTT实验证明,Chk1蛋白表达缺失导致MGC803细胞增殖活性下降,其增殖抑制率为38.0%(t=26.797,P<0.05);Chk2蛋白表达缺失对细胞增殖无明显抑制作用(t=6.098,P>0.05)。Chk1基因沉默后细胞周期在G0/G1期发生停滞,但并不引发凋亡,而Chk2基因沉默后对细胞周期影响不明显。结论:Chk1基因沉默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。OBJECTIVE：To investigate the effect of Chk1/2 gene silencing on the cell viability of human gastric cancer cell line MGC803,and explore the role of Chk1 and Chk2 in cell cycle.METHODS：The siRNA targeting at Chk1 or Chk2 gene was transfected into MGC803 cells.The protein expression of Chk1 and Chk2 was detected by Western blot.Cell viability and cell cycle rates were determined by MTT assay and FCM.RESULTS：The gray values of Chk1 and Chk2 protein were 0.09±0.04 and 0.12±0.01,both o which were lower than those of untransfected group（0.39±0.09） and liposome transfected group（0.38±0.17,P0.05）.However,there was no difference between untransfected group and liposome transfected group （P=0.458）.The Chk1 and Chk2 expression at protein level in Chk1 and Chk2 siRNA-transfected group was reduced 85.0% and 83.4% respectively.The lack of Chk1 expression resulted in decreased cellular proliferation activity,and its inhibition rate was 38.0%（t=26.767,P0.05）.However,The lack of Chk2 expression had no effect on proliferation of MGC803 cells（t=6.098,P0.05）.Further investigation revealed that the cell cycle was delayed at the G0/G1 phase with the especially remarkable arrest occurring in the cases of Chk1 siRNA,but without apoptosis.The lack of Chk2 expression was not involved in cell cycle.CONCLUSIONS：The proliferation of human gastric cancer MGC803 cells are inhibited by Chk1 gene silence.G0/G1 arrest is involved in the inhibition effect under the lack of Chk1 expression.

关 键 词：胃肿瘤 RNA干扰 细胞周期 

分 类 号：R735.2[医药卫生—肿瘤]