miR-17-5p通过调控PTEN/Akt通路介导乳腺癌细胞顺铂耐药  

MiR-17-5p mediating cisplatin resistance in breast cancer cells by regulating PTEN/Akt pathway

作　　者：赵妍 吉柳 孙成鹏 赵心宇[3] 武文慧 ZHAO Yan;JI Liu;SUN Cheng-peng;ZHAO Xin-yu;WU Wen-hui(Pharmacy Department of Chunliu Obstetrics and Gynecology Hospital,Dalian Municipal Women and Children's Medical Center,Dalian LIAONING 116000,China;College of Pharmacy,Dalian Medical University,Dalian LIAONING 116000,China;College of Laboratory Medicine,Dalian Medical University,Dalian LIAONING 116000,China)

机构地区：[1]大连市妇女儿童医疗中心(集团)春柳妇产院区药剂科,辽宁大连116000 [2]大连医科大学药学院,辽宁大连116000 [3]大连医科大学检验医学院,辽宁大连116000

出　　处：《中国新药与临床杂志》2024年第11期852-859,共8页Chinese Journal of New Drugs and Clinical Remedies

基　　金：辽宁省科学技术基金(2020-MS-256)。

摘　　要：目的探讨miR-17-5p通过调控PTEN/Akt通路介导乳腺癌细胞顺铂(DDP)耐药的作用。方法采用DDP浓度梯度递增培养MCF-7/DDP耐药细胞株,通过RT-qPCR检测MCF-7及MCF-7/DDP耐药细胞中miR-17-5p含量。将MCF-7细胞分为miR-NC组和miR-17-5p组,分别转染miR-NC及miR-17-5p mimics质粒;将MCF-7/DDP耐药细胞分为anti-miR-NC组和anti-miR-17-5p组,分别转染anti-miR-NC及anti-miR-17-5p质粒。通过RT-qPCR检测转染效率,MTT法评估转染后各组细胞对DDP的敏感性,Transwell法用于获取miR-17-5p对乳腺癌细胞侵袭能力的直接作用,流式细胞术检测细胞凋亡率。双荧光素酶报告基因实验验证miR-17-5p和PTEN的靶向关系,Western blot法检测miR-17-5p调控下细胞凋亡和PTEN/Akt通路关键蛋白的变化。结果相较MCF-7细胞,MCF-7/DDP耐药细胞中miR-17-5p表达量异常升高,PTEN表达量降低(P<0.01),PTEN作为靶基因受miR-17-5p调控。与miR-NC组相比,miR-17-5p组增殖抑制率显著降低,侵袭细胞数增加,凋亡率下降(均P<0.05),PTEN/Akt通路中抑癌蛋白PTEN、p21、p27表达降低,p-Akt308、p-Akt473、cyclin D1表达升高(P<0.01)。与anti-miR-NC组相比,anti-miR-17-5p组增殖抑制率提高,侵袭细胞数减少,凋亡率上升(均P<0.05),抑癌蛋白PTEN、p21、p27表达升高,p-Akt308、p-Akt473、cyclin D1表达降低(P<0.01)。结论敲低miR-17-5p可有效提高乳腺癌细胞的DDP敏感性,减弱其侵袭能力,诱导其凋亡,这可能与miR-17-5p对PTEN/Akt通路的调控作用有关。AIM To investigate the role of miR-17-5p in mediating cisplatin(DDP)resistance in breast cancer cells by regulating the PTEN/Akt pathway.METHODS MCF-7/DDP-resistant cell line was cultured with a gradient of increasing DDP concentrations.The content of miR-17-5p was detected in MCF-7 and MCF-7/DDP cells by RT-qPCR.MCF-7cells were divided into miR-NC and miR-17-5p groups,and transfected with miR-NC and miR-17-5p mimics plasmids,respectively.MCF-7/DDP-resistant cells were divided into anti-miR-NC and anti-miR-17-5p groups,and transfected with anti-miR-NC and anti-miR-17-5p plasmids,respectively.Transfection efficiency was defined by RT-qPCR.The drug sensitivity of DDP in each group of transfected cells was evaluated by MTT.The direct effect of miR-17-5p on the invasive ability was obtained by Transwell assay.DDP-induced apoptosis of MCF-7 and MCF-7/DDP cells after transfection was analyzed by flow cytometry.The targeting relationship between miR-17-5p and PTEN was verified by double luciferase reporter gene assay.The changes of apoptosis and key proteins of PTEN/Akt pathway under the regulation of miR-17-5p were detected by Western blot.RESULTS Compared with MCF-7 cells,the miR-17-5p expression in MCF-7/DDP-resistant cells was abnormally increased,while the PTEN expression was reduced(P<0.01).PTEN was regulated by miR-17-5p as a target gene.Compared with the miR-NC group,the proliferation inhibition rate in the miR-17-5p mimics group was significantly declined,the number of invaded cells was enhanced,and the apoptosis rate was also decreased(P<0.05),and the expressions of tumor suppressor proteins PTEN,p21 and p27 in the PTEN/Akt pathway were decreased,and the expressions of p-Akt308,p-Akt473 and cyclin D1 were increased(P<0.01).Compared with the anti-miR-NC group,the proliferation inhibition rate was increased in the anti-miR-17-5p group,the number of invaded cells was decreased,and the apoptosis rate was also increased(P<0.05),and the expression of tumor suppressor proteins PTEN,p21 and p27 was increased,and the e

关 键 词：顺铂 乳腺肿瘤 药物敏感性 细胞凋亡 PTEN/Akt通路 

分 类 号：R96[医药卫生—药理学]