LncRNA KCNQ1OT1靶向miR-9-5p对口腔癌细胞黏附性及活性的影响  

Study on the adhesion and activity mechanism of LncRNA KCNQ1OT1 targeting miR-9-5p on oral cancer cells

作　　者：王敏 庄志征 周胜男 王旭欣 王静璇 刘红悦 李松[2] WANG Min;ZHUANG Zhizheng;ZHOU Shengnan;WANG Xuxin;WANG Jingxuan;LIU Hongyue;LI Song(Department of Science and Education,Hebei Province Third Veterans Preferential Hospital,Baoding 071000,Hebei,China;Department of Stomatology,Affiliated Hospital of Hebei University,Baoding 071000,Hebei,China;Department III of Rehabilitation,Hebei Province Third Veterans Preferential,Baoding 071000,Hebei,China;Department of Stomatology,Baoding First Central Hospital,Baoding 071000 Hebei,China;Department of Stomatology,Hengshui Second People's Hospital,Hengshui 053099,Hebei,China;不详)

机构地区：[1]河北省第三荣军优抚医院科教科,河北保定071000 [2]河北大学附属医院口腔科,河北保定071000 [3]河北省第三荣军优抚医院荣康三科,河北保定071000 [4]河北省第三荣军优抚医院质控科,河北保定071000 [5]保定市第一中心医院口腔科,河北保定071000 [6]衡水市第二人民医院口腔二科,河北衡水053099

出　　处：《贵州医科大学学报》2024年第7期1011-1017,共7页Journal of Guizhou Medical University

基　　金：河北省卫生健康委员会项目资助(20210409)。

摘　　要：目的探究长链非编码RNA钾电压门控通道亚家族Q成员1反向转录物1(LncRNA KCNQ1OT1)靶向miR-9-5p对口腔鳞状细胞癌细胞黏附性及活性的影响。方法体外培养口腔癌SCC25细胞,分别转染LncRNA KCNQ1OT1阴性对照、LncRNA KCNQ1OT1 siRNA、miR-9-5p阴性对照、miR-9-5p siRNA、LncRNA KCNQ1OT1 siRNA和miR-9-5p siRNA,分别记为KCNQ1OT1-NC组、KCNQ1OT1组、miR-9-5p-NC组、miR-9-5p组,KCNQ1OT1+miR-9-5p组及口腔癌组;采用RT-PCR检测各组细胞的LncRNA KCNQ1OT1及miR-9-5p水平,MTT法检测细胞吸光度(OD)值,流式细胞仪检测细胞凋亡率,Western blot检测细胞中CD44S及CD44V5蛋白表达;通过双荧光素酶报告证实LncRNA KCNQ1OT1与miR-9-5p靶向关系。结果KCNQ1OT1组LncRNA KCNQ1OT1水平低于口腔癌组(P<0.05),miR-9-5p组miR-9-5p水平低于口腔癌组(P<0.05);与口腔癌组相比,KCNQ1OT1组、miR-9-5p组、KCNQ1OT1+miR-9-5p组细胞OD值、CD44S及CD44V5蛋白降低,细胞凋亡率升高(P<0.05),且以KCNQ1OT1+miR-9-5p组效果最为显著(P<0.05);双荧光素酶报告结果显示,miR-9-5p是LncRNA KCNQ1OT1的靶基因。结论LncRNA KCNQ1OT1可通过调控miR-9-5p表达从而降低口腔鳞状细胞癌细胞黏附性,抑制口腔鳞状细胞癌细胞增殖,诱导其凋亡。Objective To explore the mechanism of long non-coding RNA potassium voltage-gated channel subfamily Q member 1 opposite strand/antisense transcript 1(LncRNA KCNQ1OT1)targeting miR-9-5p on oral squamous cell carcinoma cell adhesion and activity.Methods SCC25 cells of oral cancer were transfected with LncRNA KCNQ1OT1 negative control,LncRNA KCNQ1OT1 siRNA,miR-9-5p negative control,miR-9-5p siRNA,and LncRNA KCNQ1OT1 siRNA and miR-9-5p siRNA,respectively.They were divided as KCNQ1OT1-NC(K1-NC)group,KCNQ1OT1(K1)group,miR-9-5p-NC(m-NC)group,miR-9-5p(m9-5p)group,KCNQ1OT1+miR-9-5p(K1+m9-5p)group,and untransfected cells were divided as oral cancer group.After transfection,the levels of LncRNA KCNQ1OT1 and miR-9-5p were determined by RT-PCR.Absorbance(OD)and apoptosis rate were measured by MTT and flow cytometry,respectively;and the expressions of CD44S and CD44V5 in cells were measured by Western blot,and the targeting relationship between LncRNA KCNQ1OT1 and miR-9-5p was detected by dual lucifluciferase reports.GraphPadPrism 8.0 software was used to analyze the statistical differences.Results The LncRNA KCNQ1OT1 level in K1 group was lower than that in oral cancer group(P<0.05),and the miR-9-5p level in miR-9-5p group was lower than that in oral cancer group(P<0.05).Compared with oral cancer group,OD value,CD44S,and CD44V5 in K1,miR-9-5p,and K1+miR-9-5p groups decreased,and apoptosis rate increased(P<0.05),and the effect of the treatment for K1+miR-9-5p group was the most significant(P<0.05).Dual luciferase reports showed that miR-9-5p was the target gene of LncRNA KCNQ1OT1.Conclusions LncRNA KCNQ1OT1 may regulate the expression of miR-9-5p to reduce the adhesion,inhibit the proliferation of oral squamous cell carcinoma cells and induce the apoptosis of oral squamous cell carcinoma cells.

关 键 词：口腔肿瘤 长链非编码RNA钾电压门控通道亚家族Q成员1反向转录物1 微小RNA-9-5p 细胞黏附性 细胞活性 

分 类 号：R739.8[医药卫生—肿瘤] R361.3[医药卫生—临床医学]