Effect Of Long Non-coding RNA MUC5B-AS1 In Lung Adenocarcinoma

Objective:This study explored the clinical significance and biological significance of MUC5B-AS1 in lung cancer from clinical samples,cells and animals,and provided a new theoretical basis for further understanding the role and mechanism of lncRNA in lung cancer.Methods:1.The expression of lncRNA MUC5B-AS1 and MUC5 B in lung adenocarcinoma tissues and paired paracancer tissues was analyzed by RT-PCR and real-time quantitative RT-PCR(qRT-PCR)and their association with clinical characteristics was analyzed.And the expression correlation between lncRNA MUC5B-AS1 and MUC5 B.Combined with TCGA public database to verify the differential expression of MUC5 B gene in lung adenocarcinoma and use Kaplan-Meier to analyze the association of MUC5 B with patient prognosis.2.We synthesized MUC5B-AS1 sequence and silence sequence fragment siRNA.Overexpression vector and siRNA of the MUC5B-AS1 were transfected into the lung adenocarcinoma cell lines to construct overexpression and silencing cell models.Patients with a later TNM stage show higher MUC5B-AS1 expression than patients with an earlier TNM stage.3.The effect of MUC5B-AS1 on lung adenocarcinoma cell growth was observed by CCK-8 method.The migration and invasion activities of lung adenocarcinoma cells were observed by Transwell chamber system.4.The effect of MUC5B-AS1 on the migration of lung adenocarcinoma cells in vivo was observed by injecting the lncRNA MUC5B-AS1 overexpression cell lines into nude mice.Results:1.The expression of MUC5B-AS1 in lung adenocarcinoma tissues was significantly higher than that in normal lung tissues(t=7.532,P<0.001);Association of MUC5B-AS1 expression with related clinical features in 72 lung adenocarcinomas,and patients with a later TNM staging(phase II/III/IV)showed higher MUC5B-than patients with an earlier TNM staging(phase I)AS1 expression level(t=1.755,P=0.084).2.The expression of MUC5 B mRNA in 72 lung adenocarcinoma samples was highly positively correlated with MUC5 B mRNA(r = 0.983,P < 0.001).MUC5B-AS1 and MUC5 B mRNA also showed high correlation in the six cell lines(A549,H1299,SPCA1,H1975,H460 and HBE)(r = 0.922,P < 0.001).3.MUC5 B mRNA expression in lung adenocarcinoma tissue was higher than that in normal lung tissue(P < 0.001)in the TCGA database.The expression level of MUC5 B mRNA in stage II/III/IV patients was also higher than that in stage I patients(P < 0.05).Meta-analysis of MUC5 B mRNA expression levels in 866 lung cancer patients using Kaplan-Meier Plotter software showed that up-regulation of MUC5 B mRNA was significantly associated with poor overall survival(hazard ratio(HR)= 1.48,95 %,confidence interval(CI)= 1.11-1.97,P = 0.007)and progression-free survival(HR = 1.74,95% CI = 1.19-2.56,P = 0.004)in patients with lung adenocarcinoma.After adjusting for gender,smoking status and tumor stage of adenocarcinoma,cox proportional hazards regression analysis showed that the correlation between MUC5 B mRNA up-regulation and overall survival remained significant(HR = 1.89,95% CI = 1.06-3.38,P = 0.031).4.There was no difference in the growth between H1299,A549 overexpressing pcDNA3.1-MUC5B-AS1 cell lines and empty plasmid group(negative control group).P>0.05);Silencing MUC5B-AS1 also had no effect on the growth in A549 cells.Compared with the empty plasmid control group,the number of migration and invasion cells significantly increased in the pcDNA3.1-MUC5B-AS1 overexpression group.Silencing MUC5B-AS1 significantly reduced the migration and invasion ability in A549.5.Upregulation of MUC5B-AS1 increased the colonization of H1299 cells in the lungs in nude mice 3,4 and 5 weeks after injecting overexpressing H1299 cells with MUC5B-AS1.The number of lung metastatic nodules was significantly increased in mice injected with H1299 cells stably overexpressing MUC5B-AS1 compared to control mice(P < 0.01).Conclusions: 1.The expression of lncRNA MUC5B-AS1 in lung adenocarcinoma tissues was higher than that in matched normal lung tissues.2.MUC5B-AS1 are highly positively correlated with its antisense MUC5 B mRNA in lung adenocarcinoma tissues.The up-regulation of MUC5 B mRNA expression in lung adenocarcinoma patients is significantly associated with poor prognosis.3.LncRNA MUC5B-AS1 can promote the migration and invasion of lung adenocarcinoma cells.