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

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survivinmRNA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。

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

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.