益气小复方通过下调lncRNA HCP5表达增强三阴性乳腺癌顺铂耐药细胞株MDA-MB-231/DDP对顺铂敏感性的机制研究  

Mechanism of Yiqi Formula enhancing sensitivity of cisplatin-resistant triple negative breast cancer cell line MDA-MB-231/DDP by down-regulating lncRNA HCP5

作　　者：吴晶晶[1] 王铮 马丽娜 王冰[1] 仲芫沅 叶媚娜[1] 陈红风[1] WU Jingjing;WANG Zheng;MA Lina;WANG Bing;ZHONG Yuanyuan;YE Meina;CHEN Hongfeng(Breast Department of Chinese Medicine,Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Comprehensive Breast Health Center,Department of General Surgery,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)

机构地区：[1]上海中医药大学附属龙华医院中医乳腺科,上海200032 [2]上海交通大学医学院附属瑞金医院普外科乳腺疾病诊治中心,上海200025

出　　处：《上海中医药杂志》2024年第1期57-65,共9页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81704074);陈红风上海市名老中医学术经验研究工作室建设项目(SHGZS-2017018)。

摘　　要：目的探讨益气小复方增强三阴性乳腺癌顺铂耐药细胞株MDA-MB-231/DDP顺铂敏感性的可能机制。方法以不同浓度的顺铂(0、2、4、8、16、32、64 mg/L)分别作用于MDA-MB-231野生、耐药细胞株24 h,噻唑蓝(MTT)比色法检测细胞增殖,鉴定耐药细胞株的顺铂耐药性。上述梯度浓度的顺铂及顺铂加固定浓度(5mg/L)益气小复方分别作用于野生、耐药细胞株及长链非编码RNA人源组织相容性白细胞抗原复合物P5(LncRNAHCP5)基因过表达、敲减的野生、耐药细胞株,MTT比色法检测细胞增殖,流式细胞仪检测细胞凋亡,实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法检测HCP5mRNA表达水平,蛋白质免疫印迹(Western blot)法检测磷酸酶张力蛋白同源物(PTEN)、磷酸化蛋白激酶B(p-Akt)蛋白表达水平。结果MDA-MB-231耐药细胞株较野生细胞株顺铂耐药指数为42.4。益气小复方同顺铂联用较单独使用顺铂,对耐药细胞株的增殖抑制率及诱导细胞凋亡比率均有提高(P<0.05)。MDA-MB-231耐药细胞株较野生细胞株,lncRNAHCP5、p-Akt蛋白表达升高,PTEN蛋白表达降低(P<0.05);益气小复方可降低MDA-MB-231耐药细胞株lncRNA HCP5、p-Akt蛋白表达,提高PTEN蛋白表达(P<0.05)。结论益气小复方可能通过下调lncRNAHCP5表达而升高PTEN蛋白表达、降低p-Akt蛋白表达,进而增强MDA-MB-231耐药细胞株对顺铂的敏感性。Objective To explore the possible mechanism of Yiqi Formula enhancing cisplatin sensitivity of Triple Negative Breast Cancer cell line MDA-MB-231/DDP.Methods With different concentrations of DDP(0,2,4,8,16,32,64 mg/L)respectively acted on MDA-MB-231 wild and drug resistant cell lines for 24 h,the cell proliferation was detected by MTT method to identify the drug resistance of the drug resistant cell lines to cisplatin.The above gradient concentration of cisplatin and cisplatin plus fixed concentration(5 mg/L)of Yiqi Formula was applied to wild and drug-resistant cell lines and LncRNA HCP5(human histocompatibility leukocyte antigen complex P5)gene overexpression,knockdown wild and drug-resistant cell lines.MTT method was used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis.RT-qPCR was used to detect the expression level of HCP5 mRNA.Western blot was used to detect the expression levels of PTEN and p-Akt protein.Results The DDP resistance index of MDA-MB-231 drug resistant cell line was 42.4 compared with that of wild cell line.Compared with DDP alone,the combination of Yiqi Formula and DDP increased the rate of proliferation inhibition and apoptosis induction of drug-resistant cell lines(P<0.05).Compared with wild cell lines,MDA-MB-231 drug resistant cell lines had higher expression of lncRNA HCP5,p-Akt protein and lower expression of PTEN protein(P<0.05),while Yiqi Formula could reduce the expression of lncRNA HCP5,p-Akt protein and increase the expression of PTEN protein in MDA-MB-231 drug resistant cell lines(P<0.05).Conclusion Yiqi Formula may increase the sensitivity of MDA-MB-231 drug-resistant cell line to DDP by down-regulating the expression of lncRNA HCP5,increasing the expression of PTEN protein and decreasing the expression of p-Akt protein.

关 键 词：三阴性乳腺癌 益气小复方 顺铂耐药 中药 黄芪多糖 作用机制 

分 类 号：R285[医药卫生—中药学]