Biological Effects And Mechanism Of Long Non-coding RNA H19 In Laryngeal Squamous Cell Cancer

Background and PurposeLaryngeal squamous cell carcinoma(LSCC)which threaten human health seriously is a highly aggressive malignant cancer.long non-coding RNAs(lncRNAs),which consist of more than 200 nucleotides and have diverse molecular functions.long non-coding RNA H19 plays key role in the progression of many human cancers.However,The regulation of LSCC progression by long non-coding RNA H19 was not well understood.We therefore investigated the function and regulatory mechanism of H19 in LSCC.MethodsH19 was knockdown in LSCC cells and cell immigration,invasion and proliferation were examined.The growth of xenografts with H19 knockdown LSCC cells was analyzed.And then we first utilized star Base v2.0 to predict H19 targets based on sequence complementarity.LSCC gene expression data was extracted from public databases and Pearson correlation analysis was performed to investigate the relationship between the expression levels of mi R-148a-3p and H19.we modulated the expression of mi R-148a-3p in Hep-2 cells by transfecting vectors overexpressing mi R-148a-3p,or vectors expressing an inhibitory sequence of mi R-148a-3p(mi R-148a-3p-in).We then performed wound healing assay,transwell cell invasion Assays and MTS assay on these transfected cells.Target Scan and mi Randa database were used to predict target genes for mi R-148a-3p.Luciferase report assay was performed to confirm the prediction.we also examined both the m RNA and the protein levels of DNMT1 when mi R-148a-3p was overexpressed or inhibited.Methylation-specific PCR analysis and q PCR were performed to investigate the relationship between H19 or mi R-148a-3p and DNA methylation.ResultsIn previous study,we found that the lncRNA H19 was upregulated in LSCC.The expression levels of H19 were inversely correlated with the survival rate of LSCC patients.We further showed that Knockdown of H19 expression inhibited LSCC cell migration,invasion and proliferation.micro RNA mi R-148a-3p as an inhibitory target for H19.Overexpression of mi R-148a-3p reduced LSCC migration,invasion and proliferation cell,while inhibition of mi R-148a-3p did the opposite.The inhibition of LSCC progression induced by H19 knockdown required the activity of mi R-148a-3p.DNA methyltransferase enzyme DNMT1 as a target of mi R-148a-3p.Cellular DNA methylation levels were inhibited by mi R-148a-3p overexpression,H19 knockdown and DNMT1 knockdown.ConclusionsThe lncRNA H19 could promote LSCC progression via mi R-148a-3p and DNMT1,and that DNA methylation was an important factor involved in the regulatory mechanism.