miR-885-3p靶向AKT1对结直肠癌细胞HT-29放射敏感性的影响  

miR-885-3p regulates radiosensitivity of colorectal cancer cell HT-29by targeting AKT1

作　　者：李全营[1] 吴大鹏 顾浩[3] 贺志宽[1] 汪洋[1] 葛政 秦长江[1] 王伟[4] Li Quanying;Wu Dapeng;Gu Hao;He Zhikuan;Wang Yang;Ge Zheng;Qin Changjiang;Wang Wei(Department of General surgery,Huaihe Hospital of Henan University,Kaifeng 475000,China;Department of Radiotherapy,Huaihe Hospital of Henan University,Kaifeng 475000,China;Department of Radiotherapy,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;Department of Radiotherapy,Henan Provincial People's Hospital,Zhengzhou 450000,China)

机构地区：[1]河南大学淮河医院普通外科,开封475000 [2]河南大学淮河医院放疗科,开封475000 [3]郑州大学第一附属医院放疗科,450000 [4]河南省人民医院放疗科,郑州450045

出　　处：《中华放射医学与防护杂志》2018年第12期899-906,共8页Chinese Journal of Radiological Medicine and Protection

基　　金：河南省医学科技攻关计划项目(201601029).

摘　　要：目的 探究miR-885-3p对结直肠癌细胞HT-29放射敏感性的影响以及作用机制。方法 荧光定量PCR检测经不同剂量(0、2、4、6、8 Gy)X射线照射后HT-29细胞中miR-885-3p的表达量;建立过表达miR-885-3p细胞株,功能试验探讨其对HT-29细胞放射敏感性的影响;生物信息学预测miR-885-3p下游调控的靶基因,双荧光素酶报告基因法进一步验证;上调和下调miR-885-3p表达量探讨miR-885-3p与靶基因丝苏氨酸蛋白激酶1(AKT1)表达量的调控关系;慢病毒转染敲减AKT1表达量,观察其对HT-29细胞放射敏感性的影响;共转染miR-885-3p模拟物,探讨过表达AKT1对miR-885-3p诱导的HT-29细胞放射敏感性的影响。结果 miR-885-3p在放射诱导的HT-29细胞中表达上调(F=46.64,P<0.05);过表达miR-885-3p和敲减AKT1可通过抑制HT-29细胞存活、促进其凋亡,从而增强HT-29细胞放射敏感性(t=12.33、12.95,P<0.05),放射增敏比(SER)分别为1.602和1.946;抑制miR-885-3p可通过促进HT-29细胞存活、抑制其凋亡从而促进HT-29细胞放射抵抗(t=11.94,P<0.05),SER为0.839;AKT1是miR-885-3p下游靶基因;过表达AKT1反转miR-885-3p增强HT-29放射敏感性的作用,SER为0.680。结论 miR-885-3p通过直接靶向AKT1增加结直肠癌HT-29细胞放射敏感性,为提高临床结直肠癌放疗敏感性提供一个靶点。Objective To investigate the effect and mechanism of miR-885-3p on the radiosensitivity of colorectal cancer cell HT-29.Methods The expression of miR-885-3p in HT-29 cells irradiated with different doses (0, 2, 4, 6, 8 Gy) of X-rays was detected by qPCR. The effect of miR-885-3p in modulating cell radiosensitivity was assessed in HT-29 cells with miR-885-3p overexpression. Bioinformatics prediction and dual luciferase reporter gene assay were employed to identify the direct target gene of miR-885-3p. Relationship between miR-885-3p and target gene tyrosine kinase 1 (AKT1) was investigated via regulation of miR-885-3p expression. The effect of AKT1 on radiosensitivity in HT-29 cells was evaluated through knockdown AKT1. The effect of AKT1 on miR-885-3p-induced radiosensitivity was detected by co-transferring miR-885-3p and AKT1 gene into HT-29 cells.Results miR-885-3p expression was up-regulated in radiation-induced HT-29 cells (F=46.64, P<0.05). Over-expression of miR-885-3p and knockdown of AKT1 enhanced cell radiosensitization by inhibiting survival and promoting apoptosis (t=12.33, 12.95, P<0.05) with SER of 1.602 and 1.946, respectively. Inhibition of miR-885-3p promoted radioresistance by increasing cell survival and inhibiting apoptosis (t=11.94, P<0.05) with a SER of 0.839. AKT1 is a target gene downstream of miR-885-3p, overexpression of AKT1 reversed the effect of miR-885-3p on cell radiosensitivity with a SER of 0.680.Conclusions miR-885-3p can enhance the radiosensitivity of colorectal cancer HT-29 cells by directly targeting AKT1, which provides a target for improving the radiosensitivity of clinical colorectal cancer.

关 键 词：结直肠癌 miR-885-3p 放疗敏感性 AKT1 

分 类 号：R735.34[医药卫生—肿瘤]