麦冬皂苷B调节Shh/Gli1通路对肝细胞癌上皮间质转化及耐药性的影响

Effects of ophiopogonin B on regulation of sonic hedgehog/glioma associated oncogene homolog 1 pathway on epithelial mesenchymal transformation and drug resistance in hepatocellular carcinoma

目的探讨麦冬皂苷B对肝细胞癌上皮间质转化(epithelial mesenchymal transformation,EMT)和化疗耐药性及音猬因子(sonic hedgehog,Shh)/胶质瘤相关癌基因同源物(glioma associated oncogene homolog,Gli)1通路的影响。方法筛选麦冬皂苷B合适浓度。将HepG2细胞分为对照组、麦冬皂苷B低(A组)、中(B组)、高浓度组(C组)、麦冬皂苷B高浓度+Shh激动剂组(D组),将HepG2/ADM细胞分为HepG2/ADM组、ADM组、麦冬皂苷B组及ADM+麦冬皂苷B组。检测细胞增殖、凋亡、EMT、Shh/Gli1通路及相关蛋白表达;检测HepG2/ADM细胞增殖、凋亡及Shh、Gli1蛋白。结果与对照组相比,A、B及C组5-乙炔基-2'脱氧尿嘧啶核苷(5-ethynyl-2'-deoxyuridine,EdU)阳性率、Ki67、B细胞淋巴瘤(B cell lymphoma,Bcl)-2、神经性钙黏蛋白、波形蛋白、Shh及Gli1表达依次下降,细胞凋亡率、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)及上皮细胞钙黏蛋白表达依次上升(P<0.05);与C组相比,D组Edu阳性率、Ki67、Bcl-2、神经性钙黏附蛋白、波形蛋白、Shh以及Gli1表达上升,细胞凋亡率、Bax、上皮细胞钙黏蛋白表达下降(P<0.05);与HepG2/ADM组相比,ADM、麦冬皂苷B组EdU阳性率、Shh及Gli1表达下降,细胞凋亡率上升(P<0.05);与ADM及麦冬皂苷B组相比,ADM+麦冬皂苷B组Edu阳性率、Shh及Gli1表达下降,细胞凋亡率上升(P<0.05)。结论麦冬皂苷B通过抑制Shh/Gli1通路抑制HepG2细胞EMT,降低其化疗耐药性。

Objective To investigate the effects of ophiopogonin B on epithelial mesenchymal transformation(EMT),chemotherapy resistance and sonic hedgehog(Shh)/glioma associated oncogene homolog(Gli)1 pathway in hepatocellular carcinoma.Methods Screening suitable concentration of ophiopogonin B.The HepG2 cells were divided into control group,ophiopogonin B low(group A),medium(group B),high concentration group(group C)and ophiopogonin B high concentration+Shh agonist group(group D).The HepG2/adriamycin(ADM)cells were divided into HepG2/ADM group,ADM group,ophiopogonin B group and ADM+ophiopogonin B group.HepG2 cell proliferation,apoptosis,EMT,Shh/Gli1 pathways and related protein expression were detected.HepG2/ADM cells were treated to detect cell proliferation,apoptosis,Shh and Gli1 proteins.Results Compared with the control group,5-ethynyl-2'-deoxyuridine(EdU)positive rate,the expressions of Ki67,B cell lymphoma(Bcl)-2,neural cadherin,vimentin,Shh and Gli1 in groups A,B and C decreased successively,while the apoptosis rate,Bcl-2 associated X protein(Bax)and cadherin expression in epithelial cells increased successively(P<0.05).Compared with group C,Edu positive rate,Ki67,Bcl-2,neural cadherin,vimentin,Shh and Gli1 expressions in group D were increased,while apoptosis rate,Bax and cadherin expression in epithelial cells were decreased(P<0.05).Compared with HepG2/ADM group,EdU positive rate,Shh and Gli1 expressions in ADM and ophiopogonin B group decreased,and apoptosis rate increased(P<0.05).Compared with ADM and ophiopogonin B groups,Edu positive rate,Shh and Gli1 expressions were decreased in ADM+ophiopogonin B group,and apoptosis rate was increased(P<0.05).Conclusion Ophiopogonin B inhibits EMT of HepG2 cells by inhibiting Shh/Gli1 pathway and reducing chemotherapy resistance.