复方苦参注射液联合奥沙利铂对人肝癌细胞株SMMC-7721增殖与凋亡的影响

The effect of Fufangkushen injection alone and the combined effect with oxaliplatin for antiproliferative and apoptotic action on human hepatoma cell line SMMC-7721

目的通过观察复方苦参注射液、奥沙利铂(OXA)及其两药联合对人肝癌细胞株SMMC-7721增殖与凋亡的影响,探讨中药与化疗药物联合应用的协同增效作用。方法采用MTT法观察复方苦参注射液、OXA及其两药联合对SMMC-7721细胞增殖的影响;应用流式细胞术分析对细胞周期的影响;采用原位末端标记法(TUNEL)和荧光染色法观察对细胞凋亡的作用。结果复方苦参注射液和OXA对SMMC-7721细胞均有明显的抑制增殖作用,该作用随着药物作用时间的延长、剂量的增加而逐渐增强(P<0.01);两药联合表现为相加或协同作用,与单药比较,有显著性差异(P<0.05)。TUNEL法检测结果表明,复方苦参注射液(25μl/ml)、OXA(1μg/ml)以及两药联合作用于SMMC-7721细胞后均可诱导肝癌细胞凋亡,凋亡指数(AI)分别为25.2%、16.2%和36.3%,联合组明显优于单药组(P<0.05)。流式细胞术分析显示,复方苦参注射液、OXA均可使细胞被阻滞于G0/G1期,两药联合较单药更为显著(P<0.05)。结论复方苦参注射液、OXA均能抑制人肝癌细胞株SMMC-7721的增殖和诱导细胞凋亡,将细胞阻滞于G0/G1期,两药联合后作用加强,提示复方苦参注射液和OXA联合应用治疗肝癌可能会起到协同增效的作用。

Objective By observing the effects of Fufangkushen injection and oxaliplatin alone and their combined effects for cell proliferation and apoptosis on human hepatoma cell lines SMMC-7721 in vitro, the effect combination of chemotherapy with traditional medicine to cure hepatocarcinoma is investigated. Methods MTT-test was used to estimate the inhibition of proliferation on SMMC-7721; flow cytometry was applied to analyze the distribution of cell cycle; fluorescence microscope and TUNEL method were employed to observe the cell apoptosis. Results Fufangkushen injection and oxaliplatin could markedly inhibit the cell proliferation, and the effect depended on exposure time and dose （P 〈 0.01 ） ; when they worked combinedly, the effect was additive and synergistic, which had statistical significance comparing with Fufangkushen injection and oxaliplatin alone. TUNEL method showed that Fufangkushen injection （25μl/ml）, oxaliplatin （1μg/ml）, as well as the two combined, could induce the apoptosis of SMMC-7721 ; the apoptosis index （AI） were 25. 2%, 16. 2% and 36. 3% respectively（P 〈0. 05）, which were obvious that the combined group significantly excelled the single work. Flow cytometry showed the proportion of cells increased at G0/G1 phase while it decreased at S phase, indicating that drugs blocked cells at G0/G1 phase. The effect of pair work was clearly superior to the single drug （ P 〈 0. 05 ）. Conclusion Fufangkushen injection and oxaliplatin can inhibit the proliferation, induce the apoptosis of SMMC-7721 cell lines, and block the cell cycle at G0/G1 phase. It is suggested that when those two drugs combined, a synergistic effect may be gotten for the treatment of liver cancer on the liver cells.