miR-302a对结直肠癌细胞奥沙利铂化疗敏感性的影响及机制  

Role of miR-302a in oxaliplatin-resistance of colorectal cancer

作　　者：孙丽娜[1] 孙利慧[1] 杜风[1] 汪鑫[1] 陈萍[1] 赵晓迪[1] 卢瑗瑗[1] 王新[1] Sun Lina;Sun Lihui;Du Feng;Wang Xin;Chen Ping;Zhao Xiaodi;Lu Yuanyuan(State Key Laboratory of Cancer Biology,Xijing Hospital of Digestive Disease, Forth Military Medical University, Shaanxi Xi＇an 710032, China.)

机构地区：[1]肿瘤生物学国家重点实验室第四军医大学西京消化病医院,陕西西安710032

出　　处：《现代肿瘤医学》2018年第9期1319-1322,共4页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:81472701;81673038;81572929;81602641)

摘　　要：目的:探究miR-302a对结直肠癌细胞奥沙利铂化疗敏感性的影响及机制。方法:在四株结直肠癌细胞系HT29、HCT8、SW480、SW1463中,通过转染miR-302a mimic构建miR-302a过表达模型,RT-PCR检测过表达效果;CCK-8法检测转染48 h后高表达miR-302a细胞的增殖能力及对奥沙利铂的敏感性;蛋白质印迹法检测转染后P-gp蛋白及Wnt/β-catenin通路相关蛋白MMP-7、c-Jun、c-myc、β-catenin、LEF1的变化。结果:相比正常小肠上皮细胞HIEC,miR-302a在结直肠癌细胞系HT29、HCT8、SW480、SW1463中的表达较低。转染mimic后,miR-302a的表达明显上调(P<0.001)。增殖实验发现miR-302a的上调并不影响结直肠癌细胞的增殖,而在转染miR-302a的细胞中加入奥沙利铂,miR-302a组HT29、HCT8、SW480和SW1463细胞存活率相比miR-NC组分别降低2.46、1.89、2.39、2.86倍。进一步探究miR-302a增加奥沙利铂敏感性的机制,发现miR-302a可抑制P-gp蛋白的表达,并且抑制Wnt/β-catenin通路相关蛋白MMP-7、c-Jun、c-myc、β-catenin、LEF1的表达。结论:miR-302a可增加结直肠癌细胞奥沙利铂化疗敏感性,其机制可能是通过抑制P-gp的蛋白表达,并抑制Wnt/β-catenin信号通路相关蛋白表达而实现。Objective： To investigate the role of miR-302a in oxaliplatin-resistance of colorectal cancer（ CRC）and primarily search the mechanism. Methods： HT29,HCT8,SW480,SW1463 cells were transfected with mimic to increase miR-302a expression,and RT-PCR identified transfection efficiency. CCK-8 detected CRC cells proliferation and sensitivity to oxaliplatin therapy. Western blot detected protein level of P-gp and Wnt/β-catenin pathway related protein. Results： Compared with human normal epithetial cells HIEC,CRC cells HT29,HCT8,SW480,SW1463 had a lower level of miR-302a. Expression of miR-302a could be increased siginificantly after transfecting miR-302a mimic（ P＜0. 001）. miR-302a could not influence CRC cells proliferation by CCK-8 assay,and miR-302a increased oxaliplatin-sensitivity in HT29,HCT8,SW480 and SW1463 cells. After treating HT29,HCT8,SW480 and SW1463 cells with oxaliplatin,survival rate of miR-302a group was 2. 46,1. 89,2. 39,2. 86 fold lower than that in miR-NC group. We further found miR-302a could suppress protein expression of P-gp and Wnt/β-catenin pathway related protein（ MMP-7,c-Jun,c-myc,active β-catenin,LEF1）. Conclusion： miR-302a increased oxaliplatin-sensitivity of CRC,and the possible mechanism was that miR-302a could suppress P-gp and Wnt/β-catenin pathway related protein expression.

关 键 词：miR-302a 结直肠癌 耐药 奥沙利铂 

分 类 号：R735.3[医药卫生—肿瘤]