下调乳酸脱氢酶-A诱导胃癌细胞凋亡及ROS产生的实验研究  

Down regulation of lactate dehydrogenase-A induces apoptosis and ROS production in gastric cancer cells

作　　者：徐湖波[1] 刘云华[1] 江应平 沈永祥[1] 陈友平[1] Xu Hubo;Liu Yunhua;Jiang Yingping;Shen Yongxiang;Chen Youping(Department of Gastroenterology,Tianmen First People's Hospital,Hubei Tianmen 431700,China)

机构地区：[1]天门市第一人民医院消化内科,湖北天门431700

出　　处：《现代肿瘤医学》2019年第5期757-761,共5页Journal of Modern Oncology

基　　金：湖北省自然科学基金项目(编号:2014CFC1104)

摘　　要：目的:探讨下调乳酸脱氢酶-A(lactate dehydrogenase-A,LDHA)对胃癌细胞凋亡及细胞中活性氧(reactive oxygen species,ROS)产生的影响。方法:将体外培养的胃癌SGC7901细胞随机分为对照组(未转染)、siN C组(转染control-siRNA)和siL DHA组(转染LDHA-siRNA),脂质体转染48 h后,采用逆转录-聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)和免疫印迹(Western blot)法检测转染效果,流式细胞仪检测细胞凋亡,Western blot检测凋亡相关蛋白活化的含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved cysteinyl aspartate specific proteinase 3,Cleaved Caspase-3)、Bcl-2相关X蛋白(Bax)和B淋巴细胞瘤-2基因(Bcl-2)的表达,激光扫描共聚焦显微镜法检测ROS水平,比色法分析细胞内氧化型谷胱甘肽(oxidized glutathione,GSSG)含量。结果:与对照组相比,siL DHA组细胞的凋亡率、Cleaved Caspase-3和Bax蛋白表达、ROS水平和GSSG含量均显著升高,而LDHA mRNA表达和LDHA、Bcl-2蛋白表达均明显降低,差异均有统计学意义(P <0. 05);而siN C组与对照组相比,凋亡率、Cleaved Caspase-3、LDHA、Bcl-2和Bax蛋白表达、LDHA mRNA表达、ROS水平和GSSG差异均无统计学意义(P> 0. 05)。结论:下调LDHA可诱导胃癌SGC7901细胞凋亡,其作用机制可能与细胞内ROS水平升高有关。Objective:To investigate the effect of down-regulation of lactate dehydrogenase-A(LDHA)on the apoptosis and reactive oxygen species(ROS)production of gastric cancer cells.Methods:Gastric cancer SGC7901 cells cultured in vitro were randomly divided into control group(non transfection),siNC group(transfection control-siRNA)and siLDHA group(LDHA-siRNA).After transfection of liposomes for 48 h,the transfection effect was detected by reverse transcription polymerase chain reaction(RT-PCR)and immunoblotting(Western blot),and apoptosis was detected by flow cytometry,and the expression of Cleaved Caspase-3,Bax and Bcl-2 proteins were tested by Western blot.The level of ROS was detected by laser scanning confocal microscopy,and the content of intracellular oxidized glutathione(GSSG)was analyzed by colorimetric assay.Results:Compared with the control group,the apoptosis rate,the expression of Cleaved Caspase-3 and Bax protein,the level of ROS and the content of GSSG in the siLDHA group were significantly increased,while the expression of LDHA mRNA and the expression of LDHA and Bcl-2 proteins were significantly decreased,and the differences were statistically significant(P<0.05).But there was no significant difference in Cleaved Caspase-3,LDHA,Bcl-2 and Bax proteins expression,LDHA mRNA expression,ROS level and GSSG content between siNC group and control group(P>0.05).Conclusion:Down-regulation of LDHA can induce apoptosis of gastric cancer SGC7901 cells,and its mechanism may be related to the increase of intracellular ROS level.

关 键 词：胃癌 乳酸脱氢酶-A 细胞凋亡 ROS 

分 类 号：R735.2[医药卫生—肿瘤]