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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 study the inhibitory effect and mechanism of survivin antisense oligonucleotide and tumor suppressor gene P53 on human gastric cancer cell line HS-746T. Methods The gastric cancer cell line HS-746T was treated with P53 gene and designed survivin antisense oligonucleotide (ASODN), and divided into blank control group, antisense survivin transfection group, P53 gene group, antisense survivin plus P53 co-transfection Dyeing group. The cell proliferation and cell growth rate were detected by cell counting and MTT assay. The expression of survivin mRNA and protein was analyzed by RT-PCR and Western blotting. TUNEL, Analysis of apoptotic index. Results The antisense survivin transfection group, P53 gene transfection group and co-transfection group all inhibited the growth of gastric cancer cells at the same time, and downregulated the expression of survivin mRNA and protein in gastric cancer cells. The transfection group had significantly higher effect than single drug group, Co-transfected group of gastric cancer apoptosis index higher than the other two groups. Conclusion Survivin antisense oligodeoxynucleotide combined with P53 gene inhibits the growth of human gastric cancer cells and induces apoptosis more than a single drug.