lncRNA HOTAIR靶向miR-126激活RhoA/ROCK通路促进喉癌Hep-2细胞增殖、侵袭与EMT  

lncRNA HOTAIR activates RhoA/ROCK pathway by targeting miR-126 and promotes the proliferation,invasion and EMT of Hep-2 cells in laryngeal cancer

作　　者：李登辉[1] 蔡润茁[1] 朱丽明[1] 陈冬 LI Denghui;CAI Runzhuo;ZHU Liming;CHEN Dong(Department of Otolaryngology,Jinzhou Central Hospital,Liaoning Jinzhou 121000,China)

机构地区：[1]锦州市中心医院耳鼻咽喉科,辽宁锦州121000

出　　处：《现代肿瘤医学》2022年第3期365-370,共6页Journal of Modern Oncology

基　　金：辽宁省自然科学基金(编号:20170540559)。

摘　　要：目的:探讨长链非编码RNA-Hox转录反义RNA(long non-coding RNA Hox antisense intergenic RNA,lncRNA HOTAIR)对喉癌Hep-2细胞增殖、侵袭及其上皮间质转化(epithelial-mesenchymal transition,EMT)的影响及机制。方法:利用实时荧光定量PCR(quantitative real-time PCR,RT-qPCR)检测喉癌组织、癌旁组织和Hep-2细胞中lncRNA HOTAIR、微小RNA-126(microRNA-126,miR-126)的表达量;下调lncRNA HOTAIR表达,分别通过细胞计数试剂盒(cell counting kit-8,CCK-8)、细胞侵袭实验和蛋白质印迹法(Western blot)检测Hep-2细胞增殖、侵袭和EMT能力。利用miRcode分析HOTAIR的靶基因,通过双荧光素酶报告基因实验分析lncRNA HOTAIR和miR-126之间的关系。同时上调lncRNA HOTAIR和miR-126的表达,检测lncRNA HOTAIR通过miR-126对Hep-2细胞增殖、侵袭和EMT的影响。下调miR-126表达后,Western blot检测RhoA、ROCK蛋白的表达量。Y-27632作用细胞后,检测下调miR-126对Hep-2细胞增殖、侵袭和EMT的影响。结果:在Hep-2细胞和喉癌组织中lncRNA HOTAIR的表达明显上调(P<0.01),miR-126表达明显下调(P<0.01)。下调lncRNA HOTAIR表达后,抑制了Hep-2细胞增殖、侵袭和EMT;lncRNA HOTAIR与miR-126具有靶向和负调控关系;下调miR-126表达后,激活了Hep-2细胞内RhoA/ROCK通路,并且促进了Hep-2细胞增殖、侵袭和EMT。结论:上调lncRNA HOTAIR通过靶向miR-126激活RhoA/ROCK通路促进喉癌Hep-2细胞增殖、侵袭与EMT。Objective:To investigate the effects and mechanism of lncRNA HOTAIR on the proliferation,invasion and EMT of Hep-2 cells in laryngeal carcinoma.Methods:The expressions of lncRNA HOTAIR and miR-126 in laryngeal cancer tissues,adjacent tissues and Hep-2 cells were detected by RT-qPCR.After downregulating the expression of lncRNA HOTAIR,the proliferation,invasion and EMT of Hep-2 cells were detected by CCK-8,Transwell and Western blot,respectively.The target genes of HOTAIR were analyzed by miRcode,and the relationship between lncRNA HOTAIR and miR-126 was analyzed by double luciferase reporter gene assay.The expressions of lncRNA HOTAIR and miR-126 were up-regulated,and the effects of lncRNA HOTAIR through miR-126 on the proliferation,invasion and EMT of Hep-2 cells were detected.After the expression of miR-126 was down-regulated,the expressions of RhoA and ROCK proteins were detected by Western blot.The effects of down-regulation of miR-126 on the proliferation,invasion and EMT of Hep-2 cells were detected in Y-27632 treated-cells.Results:The expression of lncRNA HOTAIR was significantly up-regulated in Hep-2 cells and laryngeal carcinoma tissues(P<0.01),the miR-126 expression was significantly down-regulated in Hep-2 cells and laryngeal carcinoma tissues(P<0.01).Down-regulation of lncRNA HOTAIR inhibited the proliferation,invasion and EMT of Hep-2 cells.lncRNA HOTAIR and miR-126 had a targeted and negative regulatory relationship.RhoA/ROCK pathway in Hep-2 cells was activated after down-regulation of miR-126 expression,and the proliferation,invasion and EMT of Hep-2 cells were promoted.Conclusion:Up-regulation of lncRNA HOTAIR actives RhoA/ROCK pathway by targeting miR-126 to promote proliferation,invasion and EMT of Hep-2 cells in laryngeal cancer.

关 键 词：lncRNA HOTAIR MIR-126 RhoA/ROCK通路 HEP-2细胞 增殖 侵袭 EMT 

分 类 号：R739.6[医药卫生—肿瘤]