生存素反义寡核苷酸联合 P53基因抑制胃癌细胞的生长作用

目的 研究生存素（survivin）反义寡核苷酸联合抑癌基因P53对人胃癌细胞系HS-746T的抑制作用及机制。方法 用P53基因和设计合成的survivin反义寡核苷酸（ASODN）对胃癌细胞系HS-746T进行处理，分空白对照组、反义survivin转染组，P53基因组，反义survivin加P53共转染组。采用细胞计数和四甲基偶氮唑蓝（MTT）法检测细胞增殖能力及细胞生长速度，用RT-PCR技术和Western印迹法分析survivin mRNA及蛋白质的表达情况，末端原位标记染色法（TUNEL）分析细胞凋亡指数。结果 不同时间反义survivin转染组、P53基因组和共转染组均对胃癌细胞的生长有抑制作用，且能够下凋胃癌细胞survivin mRNA和蛋白质的表达，共转染组较单独用药组效应明显增强，共转染组胃癌细胞凋亡指数高于另外两组。结论 Survivin反义寡核苷酸联合P53基因抑制人胃癌细胞的生长及诱导凋亡的作用大于单独应用一种药物。

Inhibitory effect against the gastric carcinoma cell growth by the combination of the survivin antisense oligonucleotide and P53 gene

Objective To investigate the inhibitory effect and mecha ni sm against the growth of human gastric carcinoma cell line HS-746T by the comb ination of the survivin antisense oligonucleotide (ASODN) and P53 gene and its m echanism. Methods Gastric carcinoma cell line HS-746T was treated by P53 gene and survivin antisense oligonuclerotide was designed. There were four regimen gr oups treated by different agents:ASODN alone,P53 gene alone and the combination of ASODN and P53 gene,blank control. Cell proliferative ability and cell growth were determined by cells counting and MTT. The expression of survivin mRNA and p rotein was detected by RT-PCR and Western blot respectively. Cell apoptotic in dex was detected by TUNEL. Results ASODN alone,P53 alone and the combination of ASODN and P53 could inhibit not only the growth of gastric carcinoma cell,but al so down-regulate the survivin mRNA and protein expression. The inhibitory effe ct was stronger,and the apoptosis index was higher in the combined transfection group than those in the other two single transfection groups. Conclusions The co mbination of survivin ASODN and P53 gene is more efficient to inhibit cell growt h and induce apoptosis than that of agent alone.