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目的:探讨维纳托克(Venetoclax)在HepG2、Hep3B 2个肝癌细胞株的抗癌活性。方法:对人肝癌细胞株HepG2、Hep3B细胞株进行体外培养，采用不同浓度的Venetoclax(0、3、10、30 μmol/L)处理肝癌细胞，以细胞计数试剂盒(CCK-8)检测细胞活性，0，3, 10和30 μmol/L Venetoclax作用细胞48 h，锥虫蓝拒染法检测细胞的死亡，0，5 μmol/L Venetoclax，作用24 h，流式细胞学检测细胞凋亡率，蛋白质印迹法(Western blot)检测Venetoclax诱导半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3活化，聚腺苷二磷酸核糖聚合酶(PARP)裂解。组间比较采用n t检验。n 结果:Venetoclax对HepG2、Hep3B细胞株均有较强活性抑制作用。不同浓度的Venetoclax处理肝癌细胞48、72 h，Venetoclax在HepG2细胞株中取得的半数细胞活性抑制率(ICn 50)值分别为46.5 μmol/L和14.2 μmol/L；在Hep3B细胞的ICn 50值分别为24.6 μmol/L和11.2 μmol/L。处理24 h，30.0 μmol/L的Venetoclax在HepG2和Hep3B细胞株中诱导57%[对照组、实验组凋亡率分别为(4.00±1.00)%、(56.67±8.51)%，n t＝-10.650，n P<0.01]和54% [对照组、实验组凋亡率分别为(5.67±1.53)%、(54.33±9.87)%，n t＝-8.440，n P<0.01]的肝癌细胞发生凋亡，并能诱导肝癌细胞的Caspase-3活化，PARP裂解，差异均有统计学意义。用0、3、10和30 μmol/L的Venetoclax处理48 h，在HepG2细胞株中可诱导3%(3.26±1.71)%，12%[(12.36±1.99)%，(n t＝-12.36，n P<0.05)]，32%[(32.38±4.57)%，(n t＝-10.350，n P<0.01)]和67%[(66.71±6.29)%，(n t＝-16.86，n P<0.01)]的细胞死亡，差异均有统计学意义；在Hep3B细胞株诱导2%(1.91±0.68)%，16%[(15.72±3.40)%，(n t＝-6.890，n P<0.05)]，42%[(42.39±6.86)%，(n t＝-10.180，n P<0.01)]和73%[(73.32±2.69)%，n t＝-44.490，n P<0.01]的细胞死亡，差异均有统计学意义。应用半胱氨酸蛋白酶(Caspase)抑制剂预处理，Venetoclax对HepG2和Hep3B细胞的死亡率分别从56%(56.71±6.29)%降低至11%(10.70±1.43)%，(n t＝12.350，n P<0.01)和68%(68.05±10.16)%至12%(12.7±6.12)%，(n t＝8.080，n P<0.01)，差异有统计学意义。n 结论:Venetoclax能够通过诱导Caspase的活化，诱导肝癌细胞发生凋亡，并对肝癌细胞进行杀伤。“,”Objective:To investigate biological activity of Venetoclax in two hepatocellular carcinoma (HCC) cell lines of HepG2 and Hep3B and the potential usefulness of this new drug in HCC.Methods:Liver cancer cell lines of HepG2 and Hep3B were treated with Venetoclax. The cell counting kit-8 (CCK-8) assay was used to examine the cell viability inhibition. The Trypan blue exclusion assay was used to examine the cell death induction. Annexin V staining was used to examine apoptosis, and Western blotting was used to examine cysteinyl aspartate-specific protease (Caspase) activation and poly ADP-ribose polymerase (PARP) cleavage. A Caspase inhibitor was used to investigate the Caspase-dependency of Venetoclax-induced apoptotic cell death.Results:Venetoclax potently inhibited cell viability in HCC HepG2 and Hep3B cell lines in a dose-dependent manner. Venetoclax achieved half maximal inhibitory concentration (ICn 50) values of 46.5 μmol/L and 14.2 μmol/L, respectively, in HepG2 cell line for 48- and 72-h treatment, and achieved IC n 50 values of 24.6 μmol/L and 11.2 μmol/L, respectively, in Hep3B cell line. Venetoclax at 30 μmol/L induces 57% [(4.00±1.00)% vs. (56.67±8.51)%, n t=-10.650, n P<0.01] and 54% [(5.67±1.53)% vs. (54.33±9.87)%,n t=-8.440, n P<0.01] cells positively stained with Annexin-V-Fluorescein Isothiocyanate (FTIC) in HepG2 and Hep3B cell lines, respectively after 24 h-treatment. Venetoclax also triggers activation of caspase-3 and PARP cleavage in both cell lines. Venetoclax at 0, 3, 10 and 30 μmol/L triggers 3% (3.26±1.71)%, 12% [(12.36±1.99)%,n t=-12.360, n P<0.05], 32% [(32.38±4.57)%,n t=-10.350, n P<0.01] and 67% [(66.71±6.29)%,n t=-16.860, n P<0.01] cell death, respectively, in HepG2 cell line, and 2% (1.91±0.68)%, 16% [(15.72±3.40)%,n t=-6.890, n P<0.05], 42% [(42.39±6.86)%,n t=-10.180, n P<0.01] and 73% [(73.32±2.69)%,n t=-44.490, n P<0.01] in Hep3B cell lines. Pretreatment with a caspase inhibitor largely attenuated Venetoclax-induced cell death in both cell lines, the dead cell rate on HepG2 and Hep3B cells decreased from 56% (56.71±6.29)% to 11% (10.70±1.43)%, (n t=12.350, n P<0.01) and 68% (68.05±10.16)% to 12% (12.7±6.12)%, (n t=8.080, n P<0.01) respectively.n Conclusion:Venetoclax displays potent anticancer activity by inducing apoptosis in HCC cells.