雷公藤甲素对GSDME依赖性肝癌细胞焦亡的影响

目的:探究雷公藤甲素对肝癌细胞Bel-7402焦亡的影响及其分子机制。方法:体外培养肝癌细胞Bel-7402,CCK-8检测不同浓度的雷公藤甲素(2.5、5.0、10.0μg/mL)对肝癌细胞活力影响;PI/DAPI染色观察细胞焦亡形态变化;ELISA试剂盒检测IL-1β和IL-18释放量;Western blotting检测Caspase-3、GSDME、IL-1β和IL-18表达;转染GSDME siRNA 24 h后PI/DAPI染色检测细胞焦亡形态变化。结果:CCK-8检测发现,随着雷公藤甲素浓度增加,Bel-7402细胞活力明显降低(P<0.01);给予雷公藤甲素(10.0μg/mL)能够明显促进细胞焦亡,升高细胞上清液中IL-1β和IL-18含量,促进Caspase-3、GSDME-N、IL-1β和IL-18表达(P<0.05或P<0.01);siGSDME能够明显抑制细胞焦亡形态变化,降低细胞焦亡数目。结论:雷公藤甲素能够促进GSDME依赖性细胞焦亡,抑制肝癌细胞增殖。

Effect of Triptolide on GSDME-Dependent Hepatocarcinoma Pyroptosis

Objective:To explore the effect of triptolide on pyrocytosis of liver cancer cells Bel-7402 and its molecular mechanism.Methods:BEL-7402 cells were cultured in vitro.CCK-8 was used to detect the effects of different concentrations of triptolide(2.5,5.0,10.0μg/mL)on the viability of BEL-7402 cells.PI/DAPI staining was used to observe the morphological changes of pyroptosis.ELISA kit was used to detect the release of IL-1βand IL-18.Western blotting was used to detect the expression of Caspase-3,GSDME,IL-1βand IL-18.PI/DAPI staining was used to detect the morphological changes of cell pyrolysis 24 h after transfection of GSDME siRNA.Results:CCK-8 assay showed that with the increase of triptolide concentration,the viability of Bel-7402 cells decreased significantly(P<0.01).Triptolide(10.0μg/mL)could significantly promote cell apoptosis,increase the contents of IL-1βand IL-18 in cell supernatant,and promote the expression of Caspase-3,GSDME-N,IL-1βand IL-18(P<0.05 or P<0.01).siGSDME could significantly inhibit the morphological changes and decrease the number of pyroptosis cells.Conclusion:Triptolide can promote GSDME-dependent pyroptosis and inhibit the proliferation of liver cancer cells.