氯化两面针碱对肝癌细胞Bel-7402自噬的影响  

Effects of nitidine chloride on autophagy of human hepatoma carcinoma bel-7402 cells

作　　者：廖艺楠 冯洁[1] Liao Yinan;Feng Jie(Pharmaceutical College,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学药学院,南宁530021

出　　处：《广西医科大学学报》2021年第5期989-993,共5页Journal of Guangxi Medical University

基　　金：广西自然科学基金资助项目(No.2017GXNSFAA198304);国家自然科学基金资助项目(No.81960706)。

摘　　要：目的:探讨氯化两面针碱(NC)对肝癌Bel-7402细胞系自噬的影响。方法:选取肝癌Bel-7402细胞株,在透射电子显微镜下观察NC处理Bel-7402细胞24 h后吞噬泡、自噬小体、自噬溶酶体等结构;Western bloting检测微管相关蛋白轻链3(LC3)和P62蛋白表达水平的改变;采用CCK-8法检测不同浓度NC(0μmol/L、1.5μmol/L、3μmol/L、6μmol/L、12μmol/L、24μmol/L)在加入或不加入自噬抑制剂氯喹(CQ)处理细胞24 h后对细胞增殖能力的影响。结果:与阴性对照组比较,Bel-7402细胞经过12μmol/L NC处理24 h后,在透射电镜显微镜下观察到细胞产生大量空泡,出现自噬小体和自噬溶酶体等结构;随着药物剂量增大,自噬相关蛋白LC3Ⅱ/LC3Ⅰ比值增加(P<0.05),P62表达降低(P<0.05);CCK-8法检测结果表明,随着NC给药剂量增加,细胞死亡增多(P<0.05),加入CQ能减轻NC导致的细胞死亡(P<0.05)。结论:NC能激活肝癌Bel-7402细胞自噬,引起Bel-7402细胞自噬性死亡。Objective:To investigate the effects of nitidine chloride(NC)on autophagy of human hepatoma carcinoma cell line Bel-7402.Methods:Bel-7402 cells were selected and the structures of phagocytes,autophagosomes and autophagosomes were observed under transmission electron microscopy(TEM)after 24 h treatment with NC.Western blot was used to assess the protein expression levels of LC3 and P62;the CCK-8 method was used to detect the cell viability of Bel-7402 cells treated with different concentration of NC(0μmol/L,1.5μmol/L,3μmol/L,6μmol/L,12μmol/L,24μmol/L)in combination with or without autophagy inhibitorchloroquine(CQ).Results:After the Bel-7402 cells were treated with 12μmol/L NC for 24 h,it was observed that a large number of vacuoles,phagocytic vesicles,autophagosomes and autophagolysosomes appeared under the TEM in Bel-7402 cells.With the increase of drug dose,the ratio of autophagy related proteins LC3 II/LC3I increased and the expression of P62 decreased(P<0.05).The CCK-8 assay indicated that the cell deathincreased as the dose of NC increased(P<0.05).CQ couldattenuate cell death caused by NC(P<0.05).Conclusion:NC can promote autophagy and cause the autophagic death in Bel-7402 cells.

关 键 词：肝癌 BEL-7402细胞 氯化两面针碱 自噬 

分 类 号：R735.7[医药卫生—肿瘤]