沉默FOXC2基因对乳腺癌耐阿霉素MCF-7/ADR细胞耐药性的逆转作用及机制  

Reversal effect and mechanism of silencing FOXC2 gene on chemotherapy resistance of doxorubicin-resistant MCF-7/ADR

作　　者：董云 邓军吉[1] 王先明[2] 易石坚[2] 屈洪波 DONG Yun;DENG Junji;WANG Xianming;YI Shijian;QU Hongbo(Department of Radiation,Weifang Hospital of Traditional Chinese Medicine,Weifang 261000,China;Department of Thyroid and Breast Surgery,South China Hospital of Shenzhen University)

机构地区：[1]潍坊市中医院放疗科,潍坊261000 [2]深圳大学附属华南医院甲乳外科

出　　处：《山西医科大学学报》2022年第5期526-531,共6页Journal of Shanxi Medical University

基　　金：湖南省科技厅临床医疗技术创新引导项目(2020SK50308)。

摘　　要：目的探讨沉默FOXC2基因对乳腺癌耐阿霉素MCF-7/ADR细胞耐药性的逆转作用及机制。方法MCF-7/ADR细胞分为未转染组、control-shRNA组和FOXC2-shRNA组,RT-PCR检测三组细胞FOXC2 mRNA表达情况,Western blot检测三组细胞FOXC2蛋白表达情况。实验分为未转染组、空脂质体组、control-shRNA组及FOXC2-shRNA组,MTT法检测吡喃阿霉素(THP-ADM)作用下MCF-7/ADR细胞的增殖抑制率及半数致死浓度(IC;);流式细胞术检测细胞凋亡率及细胞周期分布;Western blot检测FOXC2蛋白及乳腺癌耐药蛋白(BCRP)及EMT相关标志物E-钙黏蛋白(E-cadherin)及波形蛋白(Vimentin)表达变化。结果与未转染组及control-shRNA组相比,FOXC2-shRNA组中FOXC2 mRNA及其蛋白表达明显降低(P<0.05)。MTT显示FOXC2-shRNA组细胞对THP-ADM化疗敏感性增加,IC;由(6.08±1.23)μg/ml降至(0.65±0.01)μg/ml(P<0.05);同时该组细胞早期凋亡率为(20.35±0.12)%,明显高于未转染组、空脂质体组及control-shRNA组(P<0.05),且S期及G0/G1期细胞比例减少及G2/M期细胞比例增多(P<0.05);Western blot结果显示FOXC2-shRNA组细胞FOXC2、BCRP及间质性标志物Vimentin蛋白表达下降(P<0.05),而上皮性标志物E-cadherin蛋白表达增加(P<0.05)。结论通过RNA干扰沉默FOXC2基因表达可以诱导MCF-7/ADR细胞发生凋亡,能有效逆转MCF-7/ADR细胞对THP-ADM的耐药性,其可能通过调控EMT机制发挥作用。Objective To explore the reversal effect and mechanism of silencing FOXC2 gene on doxorubicin-resistant MCF-7/ADR cells.Methods MCF-7/ADR cells were divided into non-transfected group,control-shRNA group and FOXC2-shRNA group,and then FOXC2 mRNA and protein expression levels were determined by RT-PCR and Western blot,respectively.The experiments were divided into untransfected group,empty liposome group,control-shRNA group and FOXC2-shRNA group.The proliferation and the half lethal concentration(IC;)of MCF-7/ADR cells were detected under pyranoxorubicin(THP-ADM)by MTT,the apoptosis rate and the cell cycle distribution were determined by flow cytometry,and the expression of FOXC2 protein,breast cancer drug resistance protein(BCRP)and EMT-related markers E-cadherin and Vimentin was detected by Western blot.Results Compared with non-transfected group and control-shRNA group,FOXC2 mRNA and protein expression was significantly reduced in FOXC2-shRNA group(P<0.05).MTT showed that the MCF-7/ADR sensitivity to THP-ADM chemotherapy in FOXC2-shRNA group was increased,and IC;decreased from(6.08±1.23)μg/ml to(0.65±0.01)μg/ml(P<0.05).The early apoptosis rate was increased in FOXC2-shRNA group(20.35%±0.12%)compared with untransfected group,empty liposome group and control-shRNA group(P<0.05),the proportion of cells in S and G0/G1 was decreased,and the proportion of cells in G2/M phase was increased(P<0.05).Compared with untransfected group,empty liposome group and control-shRNA group,the protein expression of FOXC2,BCRP and Vimentin was decreased in FOXC2-shRNA group(P<0.05),while the expression of E-cadherin protein was increased(P<0.05).Conclusion Silencing of FOXC2 gene expression by RNA interference can induce the apoptosis of MCF-7/ADR cells and effectively reverse THP-ADM resistance in MCF-7/ADR cells by regulating the EMT.

关 键 词：乳腺肿瘤 MCF-7/ADR细胞 叉头框蛋白C2 吡喃阿霉素 化疗耐药性 

分 类 号：R737.9[医药卫生—肿瘤]