长链非编码RNA HHIP-AS1调节MAPK信号活性抑制非小细胞肺癌增殖、迁移和侵袭

LncRNA HHIP-AS1 regulates MAPK signaling pathway and suppresses proliferation,migration and invasion of non small cell lung cancer

目的探究长链非编码RNA(lncRNA)HHIP-AS1调控MAPK信号以及对非小细胞肺癌(NSCLC)增殖、迁移和侵袭的影响。方法收集2019年1月~2020年6月间NSCLC病人癌组织及癌旁组织40例,采用实时荧光定量PCR(RT-qPCR)检测NSCLC组织和细胞中HHIP-AS1的表达水平。H1299细胞分为空白对照组、阴性对照组(转染pcDNA3.1)、HHIP-AS1过表达组(转染pcDNA3.1-HHIP-AS1)。CCK-8法、伤口愈合实验和Transwell实验评估细胞增殖、迁移和侵袭能力。RT-qPCR和Western blotting检测p-MEK1/2和p-p38以及MMP-2的表达。结果肺腺癌(P<0.01)、鳞状细胞癌(P<0.01)和腺鳞癌组织(P<0.05)中HHIP-AS1的表达水平低于癌旁组织。肺癌细胞A549、H1975、HCC827和H1299中HHIP-AS1表达水平低于BEAS-2B细胞(P<0.01)。HHIP-AS1过表达组细胞增殖(48小时,P<0.05;72小时,P<0.01),迁移(P<0.01)和侵袭(P<0.01)能力均低于阴性对照组。另外,HHIP-AS1过表达组H1299细胞中p-MEK1/2(P<0.01)、p-p38(P<0.01)和MMP-2(P<0.01)的表达水平低于阴性对照组。结论过表达HHIP-AS1表达抑制肺癌细胞增殖、侵袭和迁移,同时抑制MAPK/MEK信号通路活性,在肺癌中发挥积极保护性作用。

Objective To elucidate the functions of long noncoding RNA HHIP-AS1 in proliferation,migration and invasion of non small cell lung cancer(NSCLC)and the effects on mitogen-activated protein kinase(MAPK)signaling pathway.Methods A total of 40 paired NSCLC tissues and adjacent normal tissues were collected from January 2019 to June 2020.The expression levels of HHIP-AS1 in NSCLC tissues and cell lines were detected by real time-quantitative polymerase chain reaction(RT-qPCR).H1299 cell were divided into blank control group,negative control group(transfected with pcDNA3.1)and overexpression-HHIP-AS1 group(transfected with pcDNA3.1-HHIP-AS1).CCK-8,Scratch and Transwell assays were used to determine the cell proliferation,migration and invasion.RT-qPCR and Western blotting were applied to detect the expressions of p-MEK1/2,p-p38,and MMP-2.Results The expressions of HHIP-AS1 in adenocarcinoma(P<0.01),squamous cell carcinoma(P<0.01)and adenosquamous carcinoma(P<0.05)tissues were decreased compared with adjacent tissues.The expressions of HHIP-AS1 in lung cancer cells A549,H1975,HCC827 and H1299 were decreased compared with BEAS-2 B cell(P<0.01).Compared with Negative control group,the proliferation(48 h,P<0.05;72 h,P<0.01),migration(P<0.01)and invasion(P<0.01)abilities of H1299 cell were reduced in overexpression-HHIP-AS1 group.Besides,the expression levels of p-MEK1/2(P<0.01),p-p38(P<0.01)and MMP-2(P<0.01)of H1299 cells in overexpression-HHIP-AS1 group were decreased as compared with negative control group.Conclusions Replenishing of HHIP-AS1 suppressed proliferation,migration and invasion of lung cancer cell,inhibited the activation of MAPK/MEK signaling pathway,and exerted promoting effects on lung cancer.