人参皂苷在治疗因CLDN18-ARHGAP26表达引起的耐化疗药治疗过程中的抗肿瘤作用  

Anti-tumor effct of ginsenosides in the treatment of chemotherapeutic drugs resistance induced by the expression of CLDN18-ARHGAP26 fusion

作　　者：李靖[1] 谢波[1] 汪虎[1] 张晨嵩 贾建光[1] 潘成武[1] 马家驰 Li Jing;Xie Bo;Wang Hu;Zhang Chensong;Jia Jianguang;Pan Chengwu;Ma Jiachi(Dept of Oncology, The First Affiliated Hospital of Bengbu Medical College,Bengbu 233004)

机构地区：[1]蚌埠医学院第一附属医院肿瘤外科,蚌埠233004

出　　处：《安徽医科大学学报》2022年第1期111-116,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽省高校自然科学研究项目(编号:KJ2019A0368)。

摘　　要：目的探究胃癌细胞中CLDN18-ARHGAP26融合突变基因引起的耐化疗药的作用,并探究人参皂苷在治疗因CLDN18-ARHGAP26融合突变基因表达引起的耐化疗药治疗过程中的抗肿瘤作用。方法采用免疫磁珠抗体标记胃癌细胞系BGC-823的侧群(SP)细胞和非侧群(NSP)细胞,选出NSP细胞转染过表达CLDN18-ARHGAP26融合突变基因的慢病毒载体。用qPCR检测细胞中CLDN18-ARHGAP26融合突变基因和三磷酸腺苷结合转运蛋白G超家族成员2(ABCG2)mRNA水平的表达。用Western blot检测转染上皮-间质转化(EMT)相关蛋白E-Cadherin、Vimentin的表达。用CCK-8检测转染细胞对化疗药奥沙利铂的敏感性。用CCK-8检测人参皂苷对转染细胞耐药性的影响。人参皂苷处理转染细胞后Western blot检测转染细胞的钙黏蛋白E(E-Cadherin)、波形蛋白(Vimentin)的表达。结果qPCR检测显示转染过表达CLDN18-ARHGAP26融合突变基因慢病毒载体的NSP细胞中CLDN18-ARHGAP26融合突变基因表达高于未转染组,ABCG2 mRNA表达高于未转染组(P<0.001)。Western bolt显示过表达CLDN18-ARHGAP26融合突变基因的NSP细胞中E-Cadherin蛋白表达低于未转染组(P<0.05),Vimentin蛋白表达高于未转染组(P<0.01),转染细胞对奥沙利铂的敏感性低于未转染组(P<0.05)。人参皂苷和奥沙利铂同时处理转染细胞,细胞存活率低于单纯奥沙利铂处理(P<0.05)。人参皂苷处理转染细胞后E-Cadherin蛋白表达高于未处理组(P<0.01),Vimentin蛋白表达低于未处理组(P<0.05)。结论人参皂苷能逆转胃癌细胞中CLDN18-ARHGAP26融合突变基因表达诱导的细胞EMT转化和奥沙利铂的耐药性。Objective To investigate the effect of chemotherapeutic drug resistance induced by CLDN18-ARHGAP26 fusion mutation gene in gastric cancer cells,and to investigate the sensitization effect of ginsenoside in the treatment of chemotherapeutic drug resistance caused by the expression of CLDN18-ARHGAP26 fusion mutation gene.Methods The side population(SP)cells and non-side population(NSP)cells of gastric cancer cell line BGC-823 were labeled with immunomagnetic bead antibody,and the lentiviral vector with overexpression of CLDN18-ARHGAP26 fusion mutation gene was selected for transfection with NSP cells.qPCR was used to detect the mRNA levels of CLDN18-ARHGAP26 fusion mutation and ATP Binding Cassette Subfamily G Member 2(ABCG2).The expression of EMT-related proteins E-Cadherin and Vimentin was detected by Western blot.The sensitivity of transfected cells to oxaliplatin was detected by CCK-8.The effect of ginsenoside on drug resistance of transfected cells was detected by CCK-8.The expression of E-Cadherin and Vimentin in transfected cells was detected byWestern blot after ginsenoside treatment.Results qPCR detection showed that the expression of CLDN18-ARHGAP26 fusion mutant gene in NSP cells transfected with overexpressed CLDN18-ARHGAP26 fusion mutant gene was significantly higher than that of the non-transfected group,and the expression of ABCG2 mRNA was significantly higher than that of the non-transfected group.Western bolt showed that the expression of E-Cadherin protein in NSP cells with over-expressed CLDN18-ARHGAP26 fusion mutation gene was lower than that in the non-transfected group,the expression of Vimentin protein was higher than that in the non-transfected group,and the sensitivity of transfected cells to oxaliplatin was lower than that in the non-transfected group.The survival rate of transfected cells treated with ginsenoside and oxaliplatin was significantly lower than that treated with oxaliplatin alone.The expression of E-Cadherin protein in the transfected cells treated with ginsenoside was higher

关 键 词：人参皂苷 CLDN18-ARHGAP26融合突变基因 上皮-间质转化 耐药性 

分 类 号：R573.9[医药卫生—消化系统]