LINC01116靶向miR-9-5p调控结直肠癌细胞对奥沙利铂的耐药性  

LINC01116 regulates resistance of colorectal cancer cells to oxaliplatin by targeting miR-9-5p

作　　者：王一飞[1] 安永铸[1] 王韬[1] 李磊[1] 郑亚男 李萍 任晓东[1] WANG Yi-Fei;AN Yong-Zhu;WANG Tao;LI Lei;ZHENG Ya-Nan;LI Ping;REN Xiao-Dong(Department of General Surgery,the First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,China)

机构地区：[1]河北北方学院附属第一医院普外科,张家口075000

出　　处：《中国免疫学杂志》2021年第19期2370-2375,共6页Chinese Journal of Immunology

基　　金：2016年河北省张家口市科技局指导性计划项目(1621090D);河北省卫健委2020年度医学科学研究课题计划项目(20200520)。

摘　　要：目的:研究LINC01116对结直肠癌细胞奥沙利铂耐药性的调控作用,并探讨其分子机制。方法:用2.5、5、10、20、40、80µmol/L浓度奥沙利铂处理亲本HCT-8细胞及HCT-8/L-OHP细胞。细胞计数试剂盒(CCK-8)检测细胞活力,计算半数抑制浓度(IC50)。实时荧光定量PCR(RT-qPCR)检测HCT-8、HCT-8/L-OHP细胞中LINC01116和miR-9-5p的表达水平。将HCT-8/L-OHP细胞分为L-OHP+si-NC、L-OHP+si-LINC01116、L-OHP+miR-NC、L-OHP+miR-9-5p、L-OHP+si-LINC01116+antimiR-NC和L-OHP+si-LINC01116+anti-miR-9-5p组,CCK-8法和流式细胞术分别检测细胞活力和凋亡。双荧光素酶报告基因实验和RT-qPCR确定LINC01116与miR-9-5p之间调控关系。结果:与亲本HCT-8细胞比较,HCT-8/L-OHP细胞显示出较强的奥沙利铂耐受性(P<0.05)。与L-OHP+si-NC组比较,抑制LINC01116表达可显著提高奥沙利铂作用下HCT-8/L-OHP细胞凋亡率和增殖抑制率(P<0.05)。与L-OHP+miR-NC组比较,miR-9-5p过表达可显著提高奥沙利铂作用下HCT-8/L-OHP细胞凋亡率和增殖抑制率(P<0.05)。LINC01116靶向负性调控miR-9-5p表达。抑制miR-9-5p表达可逆转抑制LINC01116对奥沙利铂作用下HCT-8/L-OHP细胞增殖和凋亡的影响(P<0.05)。结论:抑制LINC01116表达可降低HCT-8/L-OHP细胞对奥沙利的耐药性,其机制与靶向调控miR-9-5p表达有关。Objective:To study the effect of LINC01116 on oxaliplatin resistance in colorectal cancer cells,further to explore its molecular mechanism.Methods:HCT-8 cells and HCT-8/L-OHP cells were treated with oxaliplatin at 2.5,5,10,20,40,80µmol/L concentration.Cell counting kit(CCK-8)detected cell viability to calculate the IC50.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression levels of LINC01116 and miR-9-5p in HCT-8 and HCT-8/L-OHP cells.HCT-8/L-OHP cells were divided into L-OHP+si-NC,L-OHP+si-LINC01116,L-OHP+miR-NC,L-OHP+miR-9-5p,L-OHP+si-LINC01116+anti-miR-NC and L-OHP+siLINC01116+anti-miR-9-5p group.Cell viability and apoptosis were detected by CCK-8 method and flow cytometry,respectively.The dual luciferase reporter gene assays and RT-qPCR determined the regulatory relationship between LINC01116 and miR-9-5p.Results:Compared with HCT-8 cells,HCT-8/L-OHP cells showed stronger oxaliplatin resistance(P<0.05).Compared with the L-OHP+si-NC group,the inhibition of LINC01116 could significantly increase the apoptosis rate and proliferation inhibition rate of HCT-8/L-OHP cells treated with oxaliplatin(P<0.05).Compared with the L-OHP+miR-NC group,miR-9-5p overexpression could significantly increase the apoptosis rate and proliferation inhibition rate of HCT-8/L-OHP cells treated with oxaliplatin(P<0.05).LINC01116 targeted and negatively regulated miR-9-5p expression.Inhibiting miR-9-5p could reverse the effect of LINC01116 inhibition on the proliferation and apoptosis of HCT-8/L-OHP cells treated with oxaliplatin(P<0.05).Conclusion:Inhibiting the expression of LINC01116 can reduce the resistance of HCT8/L-OHP cells to oxaliplatin,and its mechanism is related to the targeted regulation of miR-9-5p expression.

关 键 词：LINC01116 结直肠癌 奥沙利铂耐药 miR-9-5p 

分 类 号：R735.3[医药卫生—肿瘤]