LncRNA MiR143HG’s Role And Mechanism In The Development Of Lung Squamous Carcinoma

ObjectiveBy establishing stable overexpression cell lines of miRNA143HG,the effect of overexpression of miRNA143HG on the proliferation and migration of lung squamous cell(LUSC)cells was studied,and the mechanism of its influence on the specific molecular action of cells was also discussed.MethodsGene expression data and patient survival information of LUSC group(n=502)and healthy control group(n=49)were downloaded from the TCGA database(https://tcga-data.nci.nih.gov/tcga/).By using UALCAN miR143HG website(http://ualcan.path.uab.edu)analysis in the TCGA carcinoma analysis,and the corresponding expression of cancer health organization.According to the expression levels of miR143HG in TCGA LUSC samples and healthy control samples,differential expression analysis was conducted.The rank sum test was used to analyze the difference significance.The expression level of miR143HG and the survival data of LUSC patients were combined,and the samples were divided into two groups(divided by optimal separation values)according to the expression level of miR143HG,and the survival curve was estimated by Kaplan-Meier method.Finally,Log-rank test was used to analyze its significance.The diagnostic prediction effect of miR143HG in LUSC patients was evaluated using ROC(receiver operating characteristic curve).the R package ’pROC’ is used to analyze and plot ROC curves and calculate the area under the curve(AUC).The predictive effect of miR143HG on the prognosis of LUSC patients was evaluated by time ROC.R package ’timeROC’ is used to analyze and plot timeROC curves for 1,3,5 and 10 years and calculate the area under the curve(AUC)value.Gene Set Enrichment Analysis(GSEA)was used to analyze enrichment level of pathway genes in miR143HG high expression group.First,the miR143HG gene expression median was used to divide LUSC samples into two groups(high expression group and low expression group).The LUSC gene expression matrix was input into GSEA software(version:4.2.3),and the pathway gene set in the Kyoto Encyclopedia of Genes and Genomes(KEGG)was selected for concentration analysis.The prediction starbase database(http://starbase.sysu.edu.cn/index.php)found LncRNA miR143HG binding sites with miR-155.The expression of miR143HG in NCI-H520 cells and BEAS-2B cells was detected by qRT-PCR.The expression of miR-155 in NCI-H520 cells and BEAS-2B cells was detected by qRT-PCR.Overexpression was stably constructed by liposome transient transfection technique.NCI-H520 cells were transfected with LncRNA miR143HG,simultaneously transfected with LncRNA miR143HG and miR-155 overexpressed plasmid,transfected with blank plasmid(NC group),and untreated NCI-H520 cells(blank group),and divided into four groups.The overexpression level of miR143HG was detected by qRT-PCR.The effects of miR143HG on cell proliferation and migration were detected by CCK-8 and scratch assay.The Wnt signaling pathway was detected by Western blot.ResultsIn this study,bioinformatics analysis showed the expression of miR143HG in healthy tissues of corresponding cancers.We know that the expression of healthy tissue is higher than that of cancer tissue in BLCA,BRCA,CESC,COAD,LUAD,LUSC,PAAD,PRAD,READ,SARC,STAD,THYM,UCEC,etc.However,the expression of healthy tissue in KIRC is lower than that in cancer tissue.And then we did an experimental studyIt was found that miR143HG was differentially expressed in lung squamous cell carcinoma cells and normal bronchial epithelial cells,and the expression of miR143HG in lung squamous cell carcinoma cells was higher than that in normal bronchial epithelial cells,suggesting that miR143HG may be related to tumor progression.Subsequently,we found that the proliferation and migration ability of lung squamous cell cells were significantly reduced after miR143HG was overexpressed.Overexpression of miR143HG inhibits the Wnt signaling pathway.This study provides a novel gene target for the treatment of lung squamous cell carcinoma.ConclusionIn this study,bioinformatics analysis showed the expression of miR143HG in healthy tissues of corresponding cancers.We know that the expression of healthy tissue is higher than that of cancer tissue in BLCA,BRCA,CESC,COAD,LUAD,LUSC,PAAD,PRAD,READ,SARC,STAD,THYM,UCEC,etc.However,the expression of healthy tissue in KIRC is lower than that in cancer tissue.And then we did an experimental study.It was found that miR143HG was differentially expressed in lung squamous cell carcinoma cells and normal bronchial epithelial cells,and the expression of miR143HG in lung squamous cell carcinoma cells was higher than that in normal bronchial epithelial cells,suggesting that miR143HG may be related to tumor progression.Subsequently,we found that the proliferation and migration ability of lung squamous cell cells were significantly reduced after miR143HG was overexpressed.Overexpression of miR143HG inhibits the Wnt signaling pathway.This study provides a novel gene target for the treatment of lung squamous cell carcinoma.