苦参素注射液联合顺铂对人肝癌SMMC-7721细胞凋亡相关基因c-myc、bcl-2和bax表达的影响

摘要

目的 探讨苦参素注射液(MI)联合顺铂(DDP)对人肝癌SMMC-7721细胞凋亡相关基因c-myc、bcl-2和bax表达的影响.方法 分别采用MI、顺铂及MI联合顺铂干预SMMC-7721细胞.TUNEL法检测细胞凋亡率,半定量RT-PCR法检测c-myc、bcl-2和bax mRNA表达,二步法免疫组化检测c-myc、bcl-2和bax蛋白表达.结果 苦参素组、顺铂组和联合用药组细胞凋亡率显著上升,与对照组(不干预)比较差异有统计学意义(P ＜0.05或P＜0.01),联合用药具有单纯相加或协同作用；联合用药组的细胞凋亡率显著增加,bax mRNA和蛋白表达显著升高,c-myc、bcl-2 mRNA和蛋白表达显著减少,与DDP单药组比较差异有统计学意义(P＜0.05或P＜0.01).结论 苦参素注射液联合顺铂具有单纯相加或协同诱导肝癌SMMC-7721细胞凋亡作用,其机制可能与bax基因表达上调和c-myc、bcl-2基因表达下调有关.%Objective To investigate the influence of matrine injection combined with cisplatin on the expression of apoptosis associated genes c-rayc, bcl-2 and bax in human hepatocarcinoma cell line SMMC-7721. Methods Apoptotic rate was determined by TUNEL method. The expression of c-myc, bcl-2 and bax mRNA were detected by semi-quantitive RT-PCR, and the expression of c-myc, bcl-2 and bax proteins were detected by two-step immunhistochemical staining. Results The apoptotic rate was increased significantly in matrine injection, cisplatin and combined agents group respectively, compared with the control group, the difference was statistically significant ( P < 0. 05 or P < 0. 01) . The effects of combined agents were simple addition or synergistic. The apoptotic rate was increased significantly, the expression of bax mRNA and protein was increased significantly, while c-myc, bcl-2 mRNA and proteins were decreased markedly in combined agents group, compared with cisplatin group, the difference was statistically significant ( P <0. 05 or P <0.01). Conclusion Matrine injection combined with cisplatin can obviously induce apoptosis of SMMC-7721 cells with simple addition or synergistic effects, and its mechanism may be involved in up-regulating expression of bax gene and down-regulating expression of c-myc and bcl-2 genes.