氟苯达唑对肝癌Huh7细胞活力、迁移、侵袭、凋亡及自噬的影响

Effects of flubendazole on viability,migration,invasion,apoptosis and autophagy of hepatocellular carcinoma Huh7 cells

目的:探讨氟苯达唑对肝癌Huh7细胞活力、迁移、侵袭、凋亡和自噬的影响。方法:Huh7细胞分为空白对照组和氟苯达唑低、中、高浓度组,分别给予0、2、4、8μmol/L的氟苯达唑处理。采用CCK-8法观察处理24、48、72 h后的细胞活力,采用划痕实验观察处理24 h后的细胞迁移能力,采用Transwell小室检测处理48 h后的侵袭能力,同时用Annexin V-FITC/PI双染法检测细胞凋亡,并采用Western blot法检测自噬和凋亡标志蛋白的表达情况。结果:氟苯达唑可显著抑制Huh7细胞活力,且呈浓度和时间依赖性(P<0.05)。随氟苯达唑给药浓度的增加,细胞迁移率降低,侵袭细胞数减少,而细胞凋亡率升高,LC3-Ⅱ/LC3-Ⅰ比值升高,P62蛋白表达水平下降,Bax/Bcl-2比值升高(P均<0.05)。结论:氟苯达唑可抑制肝癌Huh7细胞的活力、迁移和侵袭,促进其凋亡和自噬。

Aim:To investigate the effects of flubendazole on viability,migration,invasion,apoptosis and autophagy of hepatocellular carcinoma Huh7 cells.Methods:Huh7 cells were divided into 4 groups and were treated with different concentrations(0,2,4,8μmol/L)of flubendazole.CCK-8 assay was used to detect cell viability after being treated for 24,48,and 72 hours,scratch assay was used to detect cell migration after being treated for 24 hours.After 48-hour treatment,Transwell chamber was used to detect cell invasion,Annexin V-FITC/PI double staining was used to detect cell apoptosis,and Western blot was used to detect the expressions of autophagy and apoptosis marker proteins.Results:Flubendazole significantly inhibited the viability of Huh7 cells in a concentration-and time-dependent manner(P<0.05).With the increase of flubendazole concentration,the migration rate of cells decreased,the number of invasive cells decreased,and the apoptosis rate increased(P<0.05).Western blot showed that with the increase of flubendazole concentration,the ratio of LC3-Ⅱ/LC3-Ⅰincreased,the expression level of P62 protein decreased,and the ratio of Bax/Bcl-2 increased(P<0.05).Conclusion:Flubendazole can inhibit the viability,migration and invasion of Huh7 cells,and promote their apoptosis and autophagy.