肿瘤抑制因子RUNX3对人胃癌细胞中凋亡相关基因表达的影响  

Effect of tumor suppressor factor RUNX3 on the expression of apoptosis-related genes in gastric carcinoma cells

作　　者：张春艳[1] 刘志方[2] 徐霞[2] 曾季平[2] 于晗[1] 贾继辉[1] 

机构地区：[1]山东大学医学院微生物学教研室,山东济南250012 [2]山东大学医学院生物化学与分子生物学研究所,山东济南250012

出　　处：《中国病理生理杂志》2009年第7期1249-1253,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30800406)

摘　　要：目的:研究肿瘤抑制因子人类Runt相关转录因子3(RUNX3)对人胃癌细胞BGC823中凋亡相关基因B细胞淋巴瘤/白血病基因-2(bcl-2)、bax、半胱氨酸天冬氨酸蛋白酶-3(caspase-3)、半胱氨酸天冬氨酸蛋白酶-8(caspase-8)、半胱氨酸天冬氨酸蛋白酶-9(caspase-9)表达的影响,以揭示RUNX3促进胃癌细胞凋亡的作用机制。方法:首先构建人Runx3的真核生物表达载体pcDNA3.1-Runx3,将pcDNA3.1-Runx3及空载体pcDNA3.1分别转染BGC823细胞48h后,提取细胞的总RNA和蛋白质,应用逆转录-聚合酶链反应(RT-PCR)和蛋白免疫印迹(Western blotting)分别检测转染不同载体的细胞内RUNX3的表达情况,然后用RT-PCR和West-ern blotting检测凋亡相关基因bcl-2、bax、caspase-3、caspase-8和caspase-9的表达及蛋白的表达,β-actin作为内对照。结果:我们成功构建了人Runx3的真核生物表达载体pcDNA3.1-Runx3,将其转染BGC823细胞后,RT-PCR和Western blotting结果均显示:转染pcDNA3.1-Runx3的细胞中RUNX3的表达水平明显高于转染空载体pcDNA3.1的细胞(P<0.05);转染pcDNA3.1-Runx3的细胞中bcl-2基因的表达水平明显降低,caspase-3、caspase-9基因的表达水平明显增加(P<0.05)。结论:在BGC823细胞中,RUNX3通过下调bcl-2,上调caspase-3、caspase-9的表达促进细胞的凋亡。AIM ： To investigate the effect of tumor - suppressor RUNX3 on the transcription of apoptosis - related genes bcl - 2, bax, caspase - 3, caspase - 8, caspase - 9 in human gastric carcinoma cells BGC823, and to reveal the apoptosis molecular mechanism promoted by RUNX3. METHODS： The eukaryotic expression vector of human Runx3 gene pcDNA3.1 - Runx3 was constructed, pcDNA3.1 - Runx3 and blank vector pcDNA3.1 were transfected into BGC823 cells, respectively. After 48 h, the total mRNA and protein were acquired and the expression level of Runx3 was determined by RT - PCR and Western blotting. Then, the mRNA and protein expression of bcl - 2, bax, caspase - 3, caspase - 8 and caspase - 9 was determined by RT - PCR and Western blotting.β - actin was used as a control. RESULTS ： The eukaryotic expression vector pcDNA3.1 - Runx3 was constructed successfully and transfected into BGC823 cells. RT - PCR and Western blotting confirmed that RUNX3 level was higher in pcDNA3. 1 - Runx3 transfected BGC823 cells than that in blank vector - transfected cells （ P 〈 0. 05）. The expression level of bcl - 2 gene is lower in pcDNA3. 1 - Runx3 transfect- ed BGC823 ceils than that in pcDNA3.1 transfected BGC823 cells, while the expression levels of caspase - 3 and caspase - 9 were higher in pcDNA3.1 - Runx3 - transfected BGC823 cells than those in pcDNA3.1 - transfected BGC823 cells （ P 〈 0. 05 ）. CONCLUSION ： RUNX3 promotes apoptosis of BGC823 cells by down - regulating the transcription of bcl - 2 and up - regulating the transcription of caspase - 3 and caspase - 9.

关 键 词：RUNT相关转录因子3 BGC823细胞 细胞凋亡 

分 类 号：R361.3[医药卫生—病理学]