survivin反义寡核苷酸对肝癌耐药细胞的作用及与化疗的关系

目的研究survivin反义寡核苷酸对人肝癌耐药细胞株的增殖和凋亡情况,以及对阿霉素化疗敏感性的影响。方法将人肝癌耐药细胞系SMMC-7721/ADM分为脂质体转染组、阿霉素组、正义寡核苷酸转染组、正义寡核苷酸转染 阿霉素组、反义寡核苷酸转染组、反义寡核苷酸转染 阿霉素组共6组。MTT法检测细胞相对存活率,流式细胞仪分析细胞凋亡率变化,RT-PCR和Westernblot印迹法检测细胞survivinmRNA和蛋白表达。结果survivin-ASODN作用后的人肝癌耐药细胞SMMC-7721/ADM的细胞凋亡率明显增高(P<0.05)。survivinmRNA和survivin蛋白表达:脂质体转染对照组、阿霉素组、正义寡核苷酸转染对照组、正义寡核苷酸转染对照 阿霉素组之间survivin蛋白表达无明显差异(P>0.05)。400ng/mlsurvivingASODN组和400ng/mlASODN 阿霉素组survivinmRNA较其他4组显著降低(P<0.05),但其两组之间表达无明显差异(P>0.05)。结论survivin反义寡核苷酸能降低肝癌耐药细胞survivin表达,增强人肝癌耐药细胞对阿霉素的化疗敏感性。

Antisense oligonucleotide against survivin induces apoptosis and enhances adriamycin sensitivity of SMMC-7721/ADM cells

Objective To explore the effects of survivin-specific antisense oligonucleotide(ASODN) delivered via liposome on the growth and apoptosis of drug-resistant human hepatic cancer cell line SMMC-7721/ADM and the sensitivity of the cells to adriamycin(ADM).Methods SMMC-7721/ADM cells were divided into 6 groups and treated with liposome,ADM,sense oligonucleotide(SODN),SODN ADM,400 ng/ml ASODN,and 400 ng/ml ASODN ADM group,respectively.MTT assay was used to calculate the relative survival rates of the cells,and the changes in cell apoptosis and cycle were detected with flow cytometry.RT-PCR and Western blotting were performed to detect the expressions of survivin mRNA and protein,respectively.Results The apoptotic rate of ASODN-treated cells was much higher than that of the control cells.survivin protein expression showed no significant variation between cells treated with liposome,ADM,SODN,and SODN ADM(P>0.05),whereas compared with these 4 groups,cells treated with 400 ng/ml ASODN and 400 ng/ml ASODN ADM had significantly lowered survivin mRNA expression(P<0.05),without significant differences between the latter two groups(P>0.05).SMMC-7721/ADM cells cultured in the presence of ASODN and adriamycin showed significant growth inhibition in comparison with ASODN group and ADM group.Conclusion survivin-specific ASODN can enhance the sensitivity of SMMC-7721/ADM cells to ADM by depressing survivin expression in the cells,thus improves the effect of ADM chemotherapy for liver cancer.