lncRNA KCNQ1OT1通过靶向miR-486-5p调控舌鳞癌CAL-27细胞的增殖和凋亡  

lncRNA KCNQ1OT1 regulates the proliferation and apoptosis of tongue squamous cell carcinoma CAL-27 cells by targeting miR-486-5p

作　　者：张娟 杨晋[1] 王焱[1] 高杰[1] 靳昊[1] 马素伟 Zhang Juan;Yang Jin;Wang Yan;Gao Jie;Jin Hao;Ma Suwei(Department of Stomatology,the Second Central Hospital of Baoding,Baoding Hebei 072750,China)

机构地区：[1]保定市第二中心医院口腔科,河北保定072750

出　　处：《遵义医科大学学报》2020年第6期728-734,共7页Journal of Zunyi Medical University

基　　金：河北省保定市科学技术研究与发展指导计划项目(NO:18ZF318)。

摘　　要：目的探讨lncRNA KCNQ1OT1对舌鳞癌CAL-27细胞增殖和凋亡的影响及分子机制。方法选取我院30例舌鳞癌患者的癌组织及癌旁组织标本,用实时荧光定量PCR(RT-qPCR)检测KCNQ1OT1和miR-486-5p的表达水平;CAL-27细胞分为si-NC组、si-KCNQ1OT1组、pcDNA组、pcDNA-KCNQ1OT1组、miR-NC组、miR-486-5p组、si-KCNQ1OT1+anti-miR-NC组、si-KCNQ1OT1+anti-miR-486-5p组。四甲基偶氮唑盐比色法(MTT)检测细胞增殖;蛋白质印迹(Western blot)法检测蛋白表达;流式细胞术检测细胞凋亡;双荧光素酶报告实验和RIP实验检测KCNQ1OT1和miR-486-5p的靶向关系。结果舌鳞癌组织中KCNQ1OT1表达水平升高,miR-486-5p表达水平降低(P<0.05)。抑制KCNQ1OT1表达或过表达miR-486-5p后,CAL-27细胞OD值降低,细胞凋亡率升高,CyclinD1、Bcl-2表达水平降低,p21、Bax表达水平升高(P<0.05)。KCNQ1OT1靶向调控miR-486-5p;下调miR-486-5p表达逆转了抑制KCNQ1OT1表达对舌鳞癌CAL-27细胞增殖和凋亡的影响。结论抑制KCNQ1OT1表达可通过靶向上调miR-486-5p抑制舌鳞癌CAL-27细胞的增殖,且促进细胞凋亡。Objective To explore the effect of lncRNA KCNQ1OT1 on the proliferation and apoptosis of tongue squamous cell carcinoma CAL-27 cells and its molecular mechanism.Methods 30 cases of tongue squamous cell carcinoma in our hospital were selected,real-time fluorescent quantitative PCR(RT-qPCR)was used to detect the expressions of KCNQ1OT1 and miR-486-5p.CAL-27 cells were divided into si-NC group,si-KCNQ1OT1 group,pcDNA group,pcDNA-KCNQ1OT1 group,miR-NC group,miR-486-5p group,si-KCNQ1OT1+anti-miR-NC group,and si-KCNQ1OT1+anti-miR-486-5p group.The tetramethylazolium salt colorimetric method(MTT)was used to detect cell proliferation;Western blot was used to detect protein expression;flow cytometry was used to detect apoptosis;the dual luciferase reporter experiment and RIP assay was used to detect the targeting relationship between KCNQ1OT1 and miR-486-5p.Results The expression of KCNQ1OT1 in tongue squamous cell carcinoma tissue was increased,and the expression of miR-486-5p was decreased(P<0.05).After inhibiting the expression of KCNQ1OT1 or overexpressing miR-486-5p,the MTT OD value of CAL-27 cells was decreased,the apoptosis rate was increased,the expressions of CyclinD1 and Bcl-2 were decreased,and the expressions of p21 and Bax were increased(P<0.05).KCNQ1OT1 targets and regulates miR-486-5p;the down-regulation of miR-486-5p reversed the effect of inhibiting KCNQ1OT1 on the proliferation and apoptosis of tongue squamous cell carcinoma CAL-27 cells.Conclusion Inhibiting the expression of KCNQ1OT1 can decrease the proliferation of tongue squamous cell carcinoma CAL-27 cells by targeting the up-regulation of miR-486-5p,and promote cell apoptosis.

关 键 词：lncRNA KCNQ1OT1 miR-486-5p 舌鳞癌 增殖 凋亡 

分 类 号：R739.86[医药卫生—肿瘤]