白头翁皂苷B4调控去乙酰化酶6对胃癌细胞转移和放疗敏感性的影响  

Effect of Anemoside B4 regulating deacetylase 6 on metastasis and radiosensitivity of gastric cancer cells

作　　者：刘磊 LIU Lei(Department of Splenic,Gastric and Hepatobiliary Surgery,Jinan Hospital of Integrated Traditional Chinese and Western Medicine,Jinan,Shandong 271100,China)

机构地区：[1]济南市中西医结合医院脾胃肝胆科,山东济南271100

出　　处：《安徽医药》2022年第12期2389-2394,共6页Anhui Medical and Pharmaceutical Journal

摘　　要：目的 探讨白头翁皂苷B4(AB4)对胃癌细胞增殖、迁移、侵袭及放疗敏感性的影响及其分子机制。方法 该研究起止时间为2018年4月至2019年10月。体外培养人正常胃黏膜上皮细胞GES-1与胃癌细胞HGC-27,采用不同浓度(25、50、100μmol/L)的AB4处理24 h,通过MTT法检测细胞存活率并筛选AB4适宜浓度用于后续研究。Transwell实验检测HGC-27细胞迁移及侵袭能力。细胞克隆形成实验检测AB4对HGC-27细胞放射敏感性的影响;蛋白质印迹法检测AB4对HGC-27细胞中去乙酰化酶6(SIRT6)蛋白表达的影响;干扰SIRT6表达联合AB4处理后,采用上述检测方法检测HGC-27细胞增殖、迁移、侵袭及放射敏感性;蛋白质印迹法检测DNA激活蛋白激酶催化亚基(DNA-PKcs)、DNA双链修复蛋白Rad51、DNA修复酶Ku80、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)蛋白表达水平。结果 与NC组相比,AB4处理后HGC-27细胞存活率[(100.01±9.57)%比(86.57±6.58)%、(65.45±8.45)%、(49.58±7.96)%]显著降低(P<0.05),迁移细胞数[(98.47±8.79)个比(43.57±6.53)个]与侵袭细胞数[(88.42±9.32)个比(45.56±5.13)个]显著减少(P<0.05),MMP-2、MMP-9蛋白表达水平显著降低(P<0.05),SIRT6蛋白表达水平(0.42±0.03比1.03±0.15)显著升高(P<0.05);细胞克隆形成实验显示AB4可降低HGC-27细胞存活分数(P<0.05),增加增敏比,降低Rad51、DNA-PKcs、Ku80的表达水平(P<0.05);干扰SIRT6表达联合AB4处理后,迁移细胞数[(44.25±5.52)个比(86.47±11.16)个]与侵袭细胞数[(48.56±6.29)个比(90.17±12.13)个]显著增多(P<0.05),MMP-2、MMP-9蛋白相对表达量显著升高(P<0.05),细胞存活分数显著升高(P<0.05),Rad51、DNA-PKcs、Ku80蛋白表达水平明显升高(P<0.05)。结论 白头翁皂苷B4能够通过促进SIRT6的表达进而发挥抑制胃癌细胞HGC-27增殖、迁移及侵袭的作用,并增加胃癌细胞的放射敏感性。Objective To investigate the effect of Anemoside B4 on the proliferation,migration,invasion and radio sensitivity of gastric cancer cells and its molecular mechanism.Methods The study started and ended from April 2018 to October 2019.GES-1 cells and HGC-27 cells were cultured in vitro for 24 h with different concentrations of AB4(25,50,100 μmol/L).Cell viability was determined by MTT assay and the appropriate concentration of AB4 was screened for subsequent studies.The Transwell assay was used to detect the migration and invasion of HGC-27 cells.Cell clone formation assay was used to examine the effect of AB4 on radio sensitivity of HGC-27 cells.Western blotting was used to detect the effect of AB4 on the expression of SIRT6 protein in HGC-27 cells.After interfering with SIRT6 expression in combination with AB4 treatment,the above detection methods were used to detect proliferation,migration,invasion and radiosensitivity of HGC-27 cells.The expression levels of DNA-PKcs,Rad51,DNA repair enzymes Ku80,MMP-2 and MMP-9 were detected by Western blotting.Results Compared with the NC group,the survival rate of HGC-27 cells [(100.01±9.57)% vs.(86.57±6.58)%,(65.45±8.45)%,(49.58±7.96)%] was significantly decreased after AB4 treatment(P<0.05),the number of migrated cells(98.47±8.79 vs.43.57±6.53) and the number of invasive cells(88.42±9.32 vs.45.56±5.13) were significantly decreased(P<0.05),and the expression levels of MMP-2 and MMP-9 were significantly decreased(P<0.05),the expression level of SIRT6 protein(0.42±0.03 vs.1.03±0.15) was significantly increased(P<0.05).Cell clone formation experiments showed that AB4 could decrease the survival fraction of HGC-27 cells(P<0.05),increase the sensitization ratio,and decrease the expression levels of Rad51,DNA-PKcs and Ku80(P<0.05).After interference with SIRT6 expression and AB4 treatment,the number of migrated cells(44.25±5.52 vs.86.47±11.16) and the number of invasive cells(48.56±6.29 vs.90.17±12.13) increased significantly(P<0.05),the expression levels of MMP-2

关 键 词：白头翁属 胃肿瘤 辐射耐受性 白头翁皂苷B4 基质金属蛋白酶2 基质金属蛋白酶9 去乙酰化酶6 迁移 侵袭 

分 类 号：R285[医药卫生—中药学]