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-myc,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(P0.05 or P0.01).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(P0.05 or P0.01).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.