Hedgehog信号通路介导白藜芦醇对人胆囊癌GBC-SD细胞株凋亡和侵袭的影响  

Effects of the resveratrol mediated by Hedgehog signaling pathway on the apoptosis and invasion of gallbladder cancer GBC-SD cell line

作　　者：秦洪莉 莫立军 张瞳 高明 Hong-Li QIN;Li-Jun MO;Tong ZHANG;Ming GAO(Intravenous Drug Dispensing Center,Hongxinglong Hospital,Beidahuang Group,Shuangyashan 155811,Heilongjiang Province,China;Department of Gastroenterology,Hongxinglong Hospital,Beidahuang Group,Shuangyashan 155811,Heilongjiang Province,China)

机构地区：[1]北大荒集团红兴隆医院静脉用药集中调配中心,黑龙江双鸭山155811 [2]北大荒集团红兴隆医院消化内科,黑龙江双鸭山155811

出　　处：《药物流行病学杂志》2023年第11期1250-1258,共9页Chinese Journal of Pharmacoepidemiology

摘　　要：目的 探讨白藜芦醇(RSV)对人胆囊癌GBC-SD细胞的凋亡、侵袭能力和上皮间充质转化(EMT)的影响及其发生机制。方法 体外培养GBC-SD细胞,随机分成空白组和RSV低、中、高剂量组(0,20,50,100μmol·L-1RSV)。通过Annexin V FITC/PI双染流式分析细胞凋亡情况,RT-PCR检测B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶3 (Caspase 3)mRNA表达水平,Transwell侵袭实验检测GBC-SD细胞侵袭能力,Western blotting检测EMT标志物钙黏蛋白E(E-cadherin)、钙黏蛋白N(N-cadherin)、波形蛋白(Vimentin)的表达水平和Hedgehog信号通路成员神经胶质瘤相关癌基因同源蛋白1(Gli1)、Gli2、音猬因子(Shh)的表达。再将GBC-SD细胞随机分为空质粒组、Gli1过表达质粒组、Gli1过表达质粒+RSV组,通过Western blotting检测Gli1蛋白过表达情况,Annexin V-FITC/PI双染流式和Transwell侵袭实验分别检测上述各组的凋亡水平和侵袭能力。结果 与空白组相比,RSV各剂量组细胞凋亡率、Bax和Caspase 3 mRNA表达显著升高(P <0.05或P <0.01),细胞侵袭能力显著降低(P <0.01);RSV中、高剂量组Bcl-2 mRNA表达显著降低(P <0.01)。与空白组相比,RSV高剂量组E-cadherin蛋白表达水平显著升高,N-cadherin、Vimentin、Gli1、Gli2和Shh蛋白表达水平显著降低(P <0.01)。与转染空质粒+RSV组相比,Gli1过表达质粒+RSV组细胞的凋亡率降低,侵袭能力增加(P<0.001)。结论 RSV促进GBC-SD细胞凋亡、抑制GBC-SD细胞侵袭及EMT,其作用机制可能与抑制Hedgehog信号相关。Objective To explore the effects of resveratrol(RSV)on regulating the apoptosis,invasion ability and epithelial-meshenchymal transition(EMT)of GBC-SD cells and the underlying mechanisms.Methods GBC-SD cells were cultured in vitro,and randomly divided into blank group,RSV low,medium and high dose groups(0,20,50,100μmol·L-1 RSV).Annexin V-FITC/PI double stain flow cytometry was used to analyze the cellular apoptosis,RTPCR assay was used to test the mRNA expression level of Bcl-2,Bax and Caspase 3,Transwell invasion assay was used to test the invasion ability of GBC-SD cells,and Western blotting assay was used to detect the protein expression of EMT markers(E-cadherin,N-cadherin and Vimentin)and Hedgehog signaling members(Gli1,Gli2 and Shh).GBC-SD cells were then randomly divided into blank plasmid group,Gli1 overexpression plasmid group,and Gli1 overexpression plasmid+RSV group.■e overexpression of Gli1 protein was detected by Western blotting,and the apoptosis rate and invasion ability were detected in the above group by Annexin V-FITC/PI double stain flow cytometry and transwell invasion assay respectively.Results Compared with the blank group,the apoptosis rate,Bax and Caspase 3 mRNA expression in each RSV dose group were significantly increased(P<0.05 or P<0.01),and the cell invasion ability was significantly reduced(P<0.01);the expression of Bcl-2 mRNA in the middle and high dose groups of RSV significantly decreased(P<0.01).Compared to the blank group,the protein expression level of E-cadherin was significantly increased,while the protein expression level of N-cadherin,Vimentin,Gli1,Gli2 and Shh were significantly decreased in the high-dose group of RSV(P<0.01).Compared with the transfected empty plasmid+RSV group,the apoptosis rate of cells in the Gli1 overexpression plasmid+RSV group was reduced and the invasion ability was increased(P<0.001).Conclusion RSV promotes GBC-SD cell apoptosis,inhibits GBC-SD cell invasion and EMT,and its mechanism of action may be related to the inhibition of Hedgehog signal

关 键 词：白藜芦醇 胆囊癌 GBC-SD细胞株 Hedgehog信号通路 上皮间充质转化 凋亡 侵袭 

分 类 号：R285[医药卫生—中药学]