目的 考察塔斯品碱衍生物TasD1-6对肝癌细胞株Hep3B、HepG2和BEL7402的生长抑制作用及其作用机制。方法 应用MTT法和实时细胞分析（RTCA）技术检测TasD1-6对Hep3B、HepG2和BEL7402细胞增殖的影响，筛选敏感株后采用结晶紫染色进行细胞形态学观察，根据Annexin V FITC/PI双染法和Hoechst 33258染色法考察TasD1-6对细胞凋亡的影响，采用Western blotting法检测凋亡相关蛋白表达，流式细胞仪检测细胞周期变化。结果 MTT法和RTCA结果均表明TasD1-6能明显抑制Hep3B细胞的增殖，MTT检测其半数抑制浓度（IC₅₀）值为（11.3±1.6）μmol/L，确定后续给药浓度分别为3、6、12 μmol/L；形态学观察结果表明TasD1-6高浓度下能明显抑制Hep3B细胞增殖，且细胞皱缩，形态发生明显变化；Annexin V FITC/PI双染法和Hoechst 33258染色法均表明TasD1-6可诱导细胞发生凋亡，且呈剂量依赖性；Western blotting结果表明TasD1-6可上调p53和促凋亡蛋白Bax表达，并下调抑凋亡蛋白Mcl-1表达；流式细胞仪检测细胞周期表明TasD1-6可将Hep3B细胞阻滞于G₁期。结论 TasD1-6可抑制肝癌细胞增殖，其对Hep3B细胞作用最为显著，其作用机制可能为通过上调p53和促凋亡蛋白Bax，下调抑凋亡蛋白Mcl-1诱导细胞凋亡的产生，同时将细胞周期阻滞于G₁期，抑制细胞增殖。

Objective To analyze inhibitory effect of taspine derivative TasD1-6 on human liver cancer cells of Hep3B, HepG2, and BEL7402, and to study their preliminary mechanism of anticancer action. Methods Effect of TasD1-6 on cell growth of Hep3B, HepG2, and BEL7402 were determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and real-time cellular analysis (RTCA) assay. Effect of TasD1-6 on Hep3B cell morphology was investigated with crystal violet staining. Next, Hep3B cell apoptosis was analyzed by Annexin V FITC/PI and Hoechst 33258 staining. Flow cytometry was used to determine Hep3B cell cycle. Effect of TasD1-6 on cell apoptosis protein expression in Hep3B cell was determined by Western blotting. Results MTT and RTCA assay demonstrated that TasD1-6 significantly inhibited the growth of Hep3B cell and the IC₅₀ was (11.3 ±1.6) μmol/L. Hep3B cell morphology indicated the shrinkage and obvious morphology changes. And TasD1-6 induced the Hep3B cell apoptosis in dose-dependent manner. Cell was stopped as G₁ phase by TasD1-6. Western blotting analysis showed TasD1-6 could upregulate the protein expression of p53 and Bax and downregulate Mcl-1 protein expression (P<0.05). Conclusion Taspine derivative TasD1-6 shows better inhibitory action on the growth of human liver cancer Hep3B cell than HepG2 and BEL7402 cells. At the same time, TasD1-6 can change cell cycle and induce cell apoptosis, which probably related to upregulation of p53 and Bax protein expression and downregulation of Mcl-1 protein expression.

国家自然科学基金青年项目（21305106）