miR-155-TP53INP1调节轴在结直肠癌化疗药物敏感性中的作用机制  

Mechanism of miR-155-TP53INP1 Axis in Chemosensitivity of Colorectal Cancer

作　　者：杨乐[1] 李传昌 刘小双 黄建平[1] 刘兆龙[1] YANG Le;LI Chuan-chang;LIU Xiao-shuang;HUANG Jian-ping;LIU Zhao-long(Department of general surgery,Shuguang Hospital Affiliated to Shanghai University of traditional Chinese medicine,Shanghai,201210,China)

机构地区：[1]上海中医药大学附属曙光医院普外科,上海201210

出　　处：《现代生物医学进展》2022年第15期2818-2821,2874,共5页Progress in Modern Biomedicine

基　　金：海军军医大学医学免疫学国家重点实验室开放课题(NKLMI-2017K11)。

摘　　要：目的:初步揭示miR-155通过靶向调节TP53INP1表达水平影响结直肠癌细胞对5-FU化疗敏感性。方法:将人结肠直肠癌细胞系HCT116进行培养,提取细胞总RNA后,采用miR-155逆转录特异性引物构建反转录体系进行PCR扩增,通过qRT-PCR检测miR-155在5-FU耐药细胞HCT116/FU及敏感细胞株HCT116中的表达情况;取对数生长期细胞,分别转染miR-155mim-ics、miR-155抑制剂、miR-155阴性对照后,采用CCK-8法检测miR-155对细胞5-FU药物敏感性的影响,双荧光素酶报告基因系统验证miR-155与TP53INP1的靶基因关系,Western blot检测miR-155对TP53INP1表达的影响。结果:miR-155在HCT116/Fu细胞中的表达量是HCT116细胞的7.25倍;在相同5-FU浓度时,HCT116+阴性对照的细胞生长抑制率均高于HCT116+mimics、半数抑制浓度显著低于HCT116+mimics,差异均具有统计学意义(P<0.05);TP53INP1是miR-155的靶基因,能显著降低野生型TP53INP13'-UTR的荧光素酶活性;转染miR-155 mimics后,TP53INP1的相对表达量显著下降,转染miR-155抑制剂后,TP53INP1的相对表达量显著升高,差异均具有统计学意义(P<0.05)。结论:miR-155水平升高使HCT116细胞对5-FU的敏感性降低,miR-155可能通过靶向调节TP53INP1的表达水平,从而影响结直肠癌细胞对5-FU的敏感性。Objective:To preliminarily reveal the mechanism of miR-155 affects the sensitivity of colorectal cancer cells to 5-FU chemotherapy by targeting the expression level of TP53INP1.Methods:The human colorectal cancer cell line HCT116 was cultured,and the total RNA was extracted,then the reverse transcription system was constructed with miR-155 reverse transcription specific primer for PCR amplification,the expression of miR-155 in HCT116/Fu and HCT116 was detected by qRT PCR;Logarithmic growth cells were transfected with miR-155 mimics,miR-155 inhibitor and miR-155 negative control respectively,the effect of miR-155 on 5-FU drug sensitivity was detected by CCK-8 method,and the dual-luciferase reporter assay was used to verify the relationship between miR-155 and TP53INP1,the effect of miR-155 on the expression of TP53INP1 was detected by Western blot.Results:The expression of miR-155 in HCT116/Fu cells was 7.25 times higher than that in HCT116 cells;At the same 5-FU Concentration,the rate of cell growth inhibition of HCT116+negative control was higher than that of HCT116+mimics,and the half inhibitory concentration was significantly lower than that of HCT116+mimics(P<0.05);TP53INP1 was the target gene of miR-155,which could significantly reduce the luciferase activity of wild-type TP53INP13'-UTR;After transfection of miR-155 mimics,the relative expression of TP53INP1 decreased significantly,and after transfection of miR-155 inhibitor,the relative expression of TP53INP1 increased significantly(all P<0.05).Conclusion:The increased level of miR-155 decreased the sensitivity of HCT116 cells to 5-FU,miR-155 may affect the sensitivity of colorectal cancer cells to 5-FU by targeted regulating the expression level of TP53INP1.

关 键 词：MIR-155 TP53INP1 结直肠癌 耐药 

分 类 号：R-33[医药卫生] R735.3