槐耳菌质干预TWIST介导乳腺癌紫杉醇化疗耐药蛋白及信号转导通路的机制研究  

Study on the mechanism of Huaier intervention on TWIST-mediated paclitaxel resistant-related proteins in breast cancer and its signal transduction pathway

作　　者：董岩[1] 韩雨杉 高平[1] DONG Yan;HAN Yushan;GAO Ping(Department of Medical Oncology,First Affiliated Hospital of Dalian Medical University,Dalian 116000,Liaoning,China;Clinical College,Dalian Medical University,Dalian 116000,Liaoning,China)

机构地区：[1]大连医科大学附属第一医院肿瘤内科,辽宁大连116000 [2]大连医科大学临床学院,辽宁大连1160000

出　　处：《癌症进展》2019年第20期2384-2387,2444,共5页Oncology Progress

基　　金：辽宁省教育厅项目(L2016019)

摘　　要：目的探讨槐耳菌质干预TWIST介导乳腺癌紫杉醇化疗耐药蛋白及相关信号转导通路的机制。方法采用不同浓度紫杉醇处理乳腺癌MCF-7细胞,建立紫杉醇耐药乳腺癌细胞株MCF-7/Taxol,将MCF-7/Taxol细胞分为对照组、紫杉醇组、合用组和槐耳清膏组,采用台盼蓝拒染法检测MCF-7/Taxol细胞的存活率,采用蛋白质印迹法(Western blot)检测MCF-7/Taxol细胞中TWIST、多药耐药相关蛋白(MRP)、P-糖蛋白(P-gp)的表达水平。对MCF-7/Taxol细胞进行基因瞬时转染,将转染后的MCF-7/Taxol细胞分为TWIST-shRNA(+)组和TWISTshRNA(-)组,逆转录聚合酶链反应(RT-PCR)检测转染后MCF-7/Taxol细胞中TWIST mRNA的表达水平。采用终浓度为3 mg/L的紫杉醇处理转染后的MCF-7/Taxol细胞,随后采用Western blot检测MCF-7/Taxol细胞中磷酸化蛋白激酶B(p-AKT)的表达水平。结果药物处理24、72、120 h时,对照组、紫杉醇组、合用组和槐耳清膏组MCF-7/Taxol细胞的存活率比较,差异均有统计学意义(P﹤0.01);槐耳清膏组MCF-7/Taxol细胞的存活率低于对照组、紫杉醇组和合用组(P﹤0.05);合用组MCF-7/Taxol细胞的存活率低于对照组和紫杉醇组(P﹤0.05)。药物处理120 h时,紫杉醇组MCF-7/Taxol细胞中TWIST、MRP、P-gp蛋白的相对表达量均高于对照组,槐耳清膏组MCF-7/Taxol细胞中TWIST、MRP、P-gp蛋白的相对表达量均低于紫杉醇组和合用组,槐耳清膏组MCF-7/Taxol细胞中TWIST、P-gp蛋白的相对表达量均低于对照组。TWIST-shRNA(-)紫杉醇组MCF-7/Taxol细胞中p-AKT的相对表达量为TWIST-shRNA(-)对照组的(3.29±0.15)倍,TWIST-shRNA(+)紫杉醇组MCF-7/Taxol细胞中pAKT的相对表达量为TWIST-shRNA(+)对照组的(0.85±0.21)倍。结论槐耳菌质可以抑制紫杉醇耐药乳腺癌细胞株MCF-7/Taxol的增殖,逆转其化疗耐药性,且该作用可能与TWIST介导的AKT信号转导通路有关。Objective To investigate the mechanism of Huaier intervention on TWIST-mediated paclitaxel chemotherapeutic resistance-related proteins in breast cancer and its signal transduction pathways.Method Different concentrations of paclitaxel were used to treated breast cancer MCF-7 cells,to established a paclitaxel-resistant breast cancer cell line MCF-7/Taxol.MCF-7/Taxol cells was divided into control group,paclitaxel group,combination group and Huaier group.Cell viability of paclitaxel-resistant breast cancer cell line MCF-7/Taxol was detected by trypan blue staining.Western blot was employed to detect the expression levels of TWIST,multidrug resistance-associated protein(MRP)and P-glycoprotein(P-gp)in MCF-7/Taxol cells.MCF-7/Taxol cells were transiently transfected with genes,and the MCF-7/Taxol cells after transfection were divided into TWIST-shRNA(+)group and TWIST-shRNA(-)group,the level of TWIST mRNA in MCF-7/Taxol cells after transfection was measured by reverse transcriptase polymerase chain reaction(RT-PCR).After transfection,the MCF-7/Taxol cells were treated with paclitaxel with the final concentration of 3 mg/L,and the expression level of phosphorylation protein kinase B(p-AKT)in MCF-7/Taxol cells was detected by Western blot.Result The survival rates of MCF-7/Taxol cells in control group,paclitaxel group,combination group and Huaier group after 24,72 and 120 hours of drug treatment were statistically significant different(P<0.01).The survival rate of MCF-7/Taxol cells in Huaier group was lower than that in control group,paclitaxel group and combination group(P<0.05),and the survival rate of MCF-7/Taxol cells in the combination group was lower than that in the control group and paclitaxel group(P<0.05).After 120 hours of drug treatment,the relative expression of TWIST,MRP and P-gp protein in MCF-7/Taxol cells from paclitaxel group were higher than those in control group,and the relative expressions of TWIST,MRP and P-gp in Huaier group were all lower than those in paclitaxel group and the combination group

关 键 词：槐耳菌质 紫杉醇 耐药乳腺癌 TWIST 信号转导通路 

分 类 号：R737.9[医药卫生—肿瘤]