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目的研究生存素(Survivin)反义寡核苷酸(ASODN)对胃癌细胞系HS746T的抑制作用。方法应用SurvivinASODN转染胃癌细胞,设空白、脂质体和正义链对照组,100、200和400nmol/L反义链组。电镜下观察细胞超微结构,流质细胞术、四甲基偶氮唑盐(MTT)法、逆转录聚合酶链反应(RTPCR)及Westernblot法检测转染后2～48h细胞凋亡指数(AI),生长抑制率(IR),SurvivinmRNA和蛋白表达的变化。结果转染后反义链组均能够下调SurvivinmRNA含量和蛋白表达,凋亡细胞增多。转染后24h100、200和400nmol/L反义链组AI为14.8%、19.4%及53.8%;IR(%)为45.98±2.99、50.96±3.38、72.79±2.48,反义链组高于其他对照组。结论Survivin反义寡核苷酸能够抑制胃癌细胞增殖。
Objective To study the inhibitory effect of Survivin antisense oligonucleotide (ASODN) on gastric cancer cell line HS746T. Methods The gastric cancer cells were transfected with Survivin ASODN. The blank, liposome and sense strand control groups, 100, 200 and 400 nmol / L antisense strand groups were established. Cell apoptosis index (AI) was detected at 2 ~ 48 hours after transfection by electron microscopy, cell morphology, flow cytometry, MTT assay, reverse transcriptase polymerase chain reaction (RTPCR) and Western blotting. , Growth inhibition rate (IR), Survivin mRNA and protein expression. Results After transfection, the antisense group could downregulate the expression of Survivin mRNA and protein, and the number of apoptotic cells increased. The AI of antisense group was 14.8%, 19.4% and 53.8% respectively at 24h100, 200 and 400 nmol / L after transfection; IR (%) was 45.98 ± 2.99, 50.96 ± 3.38 and 72.79 ± 2.48, group. Conclusion Survivin antisense oligonucleotide can inhibit the proliferation of gastric cancer cells.