LncRNA TTN-AS1调控miR-139-5p/PDK1/AKT/Caspase-3信号通路促进鼻咽癌的生长和转移  

LncRNA TTN-AS1 regulates the miR-139-5p/PDK1/AKT/Caspase-3 signaling pathway to promote the growth and metastasis of nasopharyngeal carcinoma

作　　者：刘春江[1] 司峰志[1] LIU Chun-jiang;SI Feng-zhi(Department of Otorhinolaryngology,Second Affiliated Hospital of Xinjiang Medical University,Urumqi Xinjiang 830092,China)

机构地区：[1]新疆医科大学第二附属医院耳鼻喉科,新疆乌鲁木齐830092

出　　处：《局解手术学杂志》2022年第6期472-478,共7页Journal of Regional Anatomy and Operative Surgery

基　　金：新疆维吾尔自治区自然科学基金(2021D01C375)。

摘　　要：目的探究lncRNA TTN-AS1对鼻咽癌生长和转移的影响及机制。方法qRT-PCR检测TTN-AS1在NP69、TW03、C666-2、CNE2细胞中的表达。将CNE2细胞分为si-NC组(转染TTN-AS1 siRNA Negative Control)、si-TTN-AS1组(转染TTN-AS1 siRNA)、si-TTN-AS1+miR-139-5p inhibitor组(共转染TTN-AS1 siRNA和miR-139-5p inhibitor)、si-TTN-AS1+PDK1组(共转染TTN-AS1 siRNA和PDK1质粒)。CCK-8检测各组细胞增殖能力,流式细胞术检测细胞凋亡水平,Transwell小室检测细胞迁移和侵袭能力,双荧光素酶报告基因实验检测lncRNA NNT-AS1与miR-139-5p的靶向关系以及miR-139-5p与PDK1的靶向关系,Western blot检测各组细胞PDK1、p-AKT、AKT、Caspase-3表达。结果TTN-AS1在鼻咽癌细胞系TW03、C666-2、CNE2中的表达显著高于鼻咽上皮细胞系NP69(P<0.05)。相比于si-NC组,si-TTN-AS1组CNE2细胞增殖能力显著降低(P<0.05),细胞凋亡水平显著较高(P<0.05),细胞迁移和侵袭数显著减少(P<0.05)。miR-139-5p为TTN-AS1的靶基因,PDK1为miR-139-5p的靶基因。相比于si-TTN-AS1组,si-TTN-AS1+miR-139-5p inhibitor组和si-TTN-AS1+PDK1组CNE2细胞增殖能力显著增加(P<0.05)、细胞迁移和侵袭数显著增加(P<0.05),细胞凋亡水平显著较低(P<0.05)。相比于si-NC组,si-TTN-AS1组CNE2细胞中PDK1蛋白表达显著下调(P<0.001),Caspase-3蛋白表达显著上调(P<0.001),p-AKT水平显著下调(P<0.001);相比于si-TTN-AS1组,si-TTN-AS1+miR-139-5p inhibitor组和si-TTN-AS1+PDK1组CNE2细胞中PDK1蛋白表达显著上调(P<0.001),Caspase-3蛋白表达显著下调(P<0.001),p-AKT水平显著上调(P<0.001)。结论LncRNA TTN-AS1可抑制鼻咽癌细胞的生长和转移,其机制为调控miR-139-5p/PDK1/AKT/Caspase-3信号通路。Objective To explore the effect and mechanism of lncRNA TTN-AS1 on the growth and metastasis of nasopharyngeal carcinoma.Methods qRT-PCR was used to detect the expression of TTN-AS1 in NP69,TW03,C666-2,and CNE2 cells.CNE2 cells were divided into the si-NC group(transfected with TTN-AS1 siRNA Negative Control),the si-TTN-AS1 group(transfected with TTN-AS1 siRNA),the si-TTN-AS1+miR-139-5p inhibitor group(co-transfected with TTN-AS1 siRNA and miR-139-5p inhibitor),and the si-TTN-AS1+PDK1 group(co-transfected with TTN-AS1 siRNA and PDK1 plasmid).CCK-8 method was used to detect the cell proliferation ability in each group;flow cytometry was used to detect the cell apoptosis level;Transwell chamber method was used to detect the cell migration and invasion ability;dual luciferase reporter gene assay was used to detect the targeting relationship between lncRNA NNT-AS1 and miR-139-5p,and the targeting relationship between miR-139-5p and PDK1;Western blot was used to detect the expression of PDK1,p-AKT,AKT,and Caspase-3 in each group.Results The expression of TTN-AS1 in the nasopharyngeal carcinoma cell lines TW03,C666-2,and CNE2 was significantly higher than that of the nasopharyngeal epithelial cell line NP69(P<0.05).Compared with the si-NC group,the proliferation ability of CNE2 cell in the si-TTN-AS1 group was significantly decreased(P<0.05),the level of cell apoptosis was significantly increased(P<0.05),and the number of cell migration and invasion was significantly decreased(P<0.05).miR-139-5p was the target gene of TTN-AS1,and PDK1 was the target gene of miR-139-5p.Compared with the si-TTN-AS1 group,the cell proliferation ability of the si-TTN-AS1+miR-139-5p inhibitor group and the si-TTN-AS1+PDK1 group was significantly increased(P<0.05),the number of cell migration and invasion was significantly increased(P<0.05),and the level of cell apoptosis was significantly decreased(P<0.05).Compared with the si-NC group,the expression of PDK1 protein of the CNE2 cells in the si-TTN-AS1 group was significantly down-regulated(

关 键 词：lncRNA TTN-AS1 鼻咽癌 生长和转移 机制 

分 类 号：R394.2[医药卫生—医学遗传学]