摘要
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survivinmRNA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,且能够下调胃癌细胞survivinmRNA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。
Objective To investigate the inhibitory effect and mechanism against the growth of human gastric carcinoma cell line HS-746T by the combination of the survivin antisense oligonucleotide （ASODN） and P53 gene and its mechanism. Methods Gastric carcinoma cell llne HS-746T was treated by P53 gene and survivin antisense oligonuclerotide was designed. There were four regimen groups 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 MTY. The expression of survivin mRNA and protein was detected by RT-PCR and Western blot respectively. Cell apoptotic index 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 also down-regulate the survivin mRNA and protein expression. The inhibitory effect was stronger, and the apoptosis index was higher in the combined transfection group than those in the other two single transfection groups. Conclusions The combination of survivin ASODN and P53 gene is more efficient to inhibit cell growth and induce apoptosis than that of agent alone.
出处
《中华胃肠外科杂志》
CAS
2005年第5期451-454,共4页
Chinese Journal of Gastrointestinal Surgery
