目的 研究大黄素对人肝癌Huh7细胞的凋亡作用及分子机制。方法 培养肝癌Huh7细胞,1、3、10、30、100 μmol/L的大黄素处理24 h,5-氟尿嘧啶(5-FU)作为阳性药,MTT法检测细胞存活率;30 μmol/L大黄素处理细胞0、3、6、24 h,光学显微镜观察细胞形态变化;Annexin V-FITC/PI双染法与流式细胞术检测细胞凋亡,Western blotting检测细胞凋亡相关蛋白的表达变化。结果 大黄素显著抑制Huh7细胞生长且呈浓度依赖性,其IC₅₀值为11.55 μmol/L。30 μmol/L大黄素处理细胞,随着药物作用时间的增加,细胞明显固缩和凝聚,大量细胞脱离培养皿底部;细胞凋亡明显;p-Akt、Bcl-2蛋白表达量减少,cleaved caspase-3蛋白表达量增加。结论 大黄素通过Akt信号途径诱导肝癌Huh7细胞凋亡。

Objective To elucidate the effect of emodin on inducing apoptosis of humam hepatoma Huh7 cells and its molecular mechanism. Methods Huh7 cells were treated with 1, 3, 10, 30 and 100 mol/L emodin for 24 h, 5-FU as positive drug. The viability of Huh7 cells was detected by MTT assay. Cells were treated with 30 μmol/L emodin for 0, 3, 6 and 24 h, and the morphological changes of cells were observed by optical microscope. The apoptosis of Huh7 cells were determined by Annexin V-FITC/PI double staining and flow cytometer. The expression levels of Akt signalling pathway proteins were determined by Western blotting. Result Emodin could significantly inhibit the proliferation of Huh7 cells in a dose-dependent manner. The IC₅₀ values were determined as 11.55 μmol/L. Cells were treated with 30 mol/L emodin, with the increasing of the time, cells significantly reduced and condensed, a large number of cells droped from the bottom of the culture dish; Emodin could induce apoptosis; emodin could significantly down-regulated p-Akt, Bcl-2 protein expression levels while up-regulated the cleaved caspase-3 expression levels in Huh7 cells. Conclusion These findings suggested that emodin can induce mitochondrial-related apoptosis on Huh7 cells by inhibiting Akt activity.

黑龙江省博士后科研启动项目(LBH-Q13132);黑龙江省自然科学基金资助项目(LC2015036);学成、引进人才科研启动计划(XYB2013-24)