鳄胆素联合阿霉素对人肝癌细胞SMMC-7721的抑制作用

Effect of Crocodile Choline Combined with Doxorubicin on Proliferation of Human Hepatocellular Carcinoma Cells SMMC-7721

研究了鳄胆素(crocodile choline)联合阿霉素(doxorubicin)对人肝癌细胞SMMC-7721的体外抑制作用.以不同浓度鳄胆素、阿霉素单用药组和两药联合用药组作用SMMC-7721细胞,应用噻唑蓝(MTT)比色法和集落形成实验检测细胞增殖抑制、流式细胞仪分析细胞周期分布、免疫印记检测凋亡相关蛋白表达.结果显示鳄胆素、阿霉素单药与两药联合均能抑制SMMC-7721细胞的增殖,且呈剂量依赖效应,两药联合有协同效应.集落形成实验中对照组、鳄胆素单用药组、阿霉素单用药组和两药联合组的克隆形成率分别为88.0%,30.8%,28.5%和1.0%,表明两药联合使克隆形成率显著降低.细胞周期分析显示药物处理组出现S期阻滞,联合组与对照组相比,S期细胞含量由12%上升到59%;各组均能诱导SMMC-7721细胞凋亡,并以两药联合组效果更佳,细胞核经Hoechst染色出现浓染致密的固缩形态和强蓝色荧光;细胞中促凋亡蛋白Bax表达上调诱导细胞凋亡.研究结果表明鳄胆素与阿霉素联合对人肝癌细胞SMMC-7721有显著的抑制作用,作用效果具有协同效应.

In this paper, we studied the effect of crocodile choline combined with doxorubicin on proliferation of human hepatocellular carcinoma cells SMMC-7721 in vitro. SMMC-7721 cells were treated with different concentrations of crocodile choline alone or com- bined with doxorubicin, and then MTT assay, colony formation assay, flow cytometry analysis, Hoechst 33258 staining, and Western blot were performed to detect relevant indicators. The results showed that crocodile choline with or without doxorubicin had inhibito- ry effect on proliferation of SMMC-7721 cells. The inhibiting rates increased with the increasing concentrations of crocodile choline and doxorubicin. The combination of the two drugs showed synergistic effect. Moreover, the results showed the colony forming rates of control group,crocodile choline group,doxorubicin group and combination group were 88.0% ,30.8%,28.5% and 1.0% ,respec- tively,indicating the combination of the two drugs could decrease the colony forming rate apparently. The results of flow cytometry analysis showed that SMMC-7721 cell cycles were blocked in S phase,the cell population of S phase increased from 12% in the con- trol group to 59% in the combination group. We found that all treatment groups could induce cell apoptosis and the combination group showed better talent. Cell nucleus concentrated and appeared granular fluorescence were observed by Hoechst 33258 staining. In the combination group, the expression of Bax protein in SMMC-7721 cells was up-regulated,inducing apoptosis of SMMC-7721 cells. This study suggests that a synergistic effect is observed between crocodile choline and doxorubicin in their inhibition of SMMC- 7721 cell proliferation and inducing apoptosis,which may give theoretical support for the treatment of human hepatocellular carcino- ma on drug combination.