Survivin反义寡核苷酸诱导SMMC-7721细胞凋亡及对化疗药物敏感性作用的研究

Antisense oligonucleotide targeting survivin induces apoptosis of SMMC-7721 cell and enhances its sensitivity to chemotherapeutic agents in vitro

目的:观察survivin反义寡核苷酸(ASODN)对SMMC-7721细胞增殖、凋亡的影响及其对化疗药物敏感性的作用。方法:设计合成特异性survivin的ASODN,脂质体转染肝细胞癌SMMC-7721细胞,透射电镜观察细胞超微结构变化;RT-PCR法检测survivin mRNA的表达变化;FCM法检测对细胞周期、凋亡及survivin蛋白表达的影响;MTT法测定survivin表达抑制前后细胞对吡柔比星、氟苷、顺铂敏感性的影响。结果:ASODN转染后细胞呈现凋亡的形态学改变,survivin mRNA和蛋白表达减弱(P<0.05),诱导SMMC-7721细胞凋亡(P<0.01),细胞周期阻滞于G2/M期(P<0.05)。ASODN转染组可增加SMMC-7721细胞对吡柔比星、氟苷、顺铂的敏感性(P<0.01)。结论:Survivin ASODN转染能下调survivin表达,诱导SMMC-7721细胞凋亡,提高对吡柔比星、氟苷、顺铂的敏感性。

Objective：To observe the effect of antisense oligonucleotide （ASODN） targeting survivin on apoptosis, proliferation and sensitivity to chemotherapeutic agents of hepatocellular carcinoma cell line SMMC-7721. Methods： Specific ASODN targeting survivin was designed and constructed, and transfected into SMMC-7721 cells mediated by oligofectamine. Cell morphological changes were examined by transmission electron microscopy. The expression of survivin mRNA was detected by RT-PCR. Cell cycle, apoptosis and the expression of survivin protein were detected by flow cytometry. Changes of sensitivity to pirarubicin, doxifluridine, and cisplatin of SMMC-7721 cells were detected by MTT assay. Results： Morphological abnormalities of cells were observed in ASODN transfected groups. The mRNA and protein expressions of survivin were significantly decreased compared with that of control group （P 〈 0.05 ）. The transfection of survivin ASODN induced significant apoptosis （P 〈 0.01 ） and increased the proportion of cells in the G2/M phase（P 〈 0.05 ）. Compared with that in control group, oligofectamine group and SODN groups, the sensitivity of ASODN groups to pirarubicin, doxifluridine, and cisplatin were significantly enhanced （P 〈 0.01 ）. Conclusion： The transfection of survivin ASODN down-regulated the mRNA and protein expressions of survivin, induces apoptosis, and enhances its sensitivity to pirarubicin, doxifluridine, and cisplatin of SMMC-7721 cells.