The Mechanism Of LncRNA H19 Promoting Cell Proliferation And Invasion In Colorectal Cancer

Background and Purpose LncRNAs,belong to non-coding RNAs(nc RNAs),without protein-coding capacity.LncRNA H19 has been found to be overexpressed in various cancers,including colorectal cancer(CRC),and may act as an oncogene.However,the mechanism by which H19 regulates CRC progression remains poorly understood.Therefore,in this study,we aimed to assess lncRNA H19 expression levels in CRC tissues,determine the effect of H19 on CRC proliferation,migration and invasion,then explore the mechanism by which lncRNA H19 regulates the proliferation,migration and invasion of CRC.Materials and Methods Lentiviral vectors were employed to overexpress lncRNA H19 in HCT116 and SW480 cells to explore the effect of lncRNA H19 in colorectal cancer cells.Then,these si RNA-H19 were transfected in HCT116 and SW480 cells.The effects of lncRNA H19 on colorectal cancer cell proliferation were analyzed via colony formation assays,cell growth curve and cell viability assay.The colorectal cancer cell migration and invasion were surveyed via Transwell migration and invasion assays.In order to understand the molecular mechanism of of lncRNA H19 on the proliferation,migration and invasion of colorectal cancer cells,as well as possible target of lncRNA H19.The Lnc Base Predicted v.2 of DIANA tools software was used and mi R-200 a was found to be the possible target combined with lncRNA H19 in colorectal cancer cells.To test this viewpoint,a series of molecular biological techniques were used to confirm that interaction relation of mi R-200 a and lncRNA H19.Then the cell growth curve and cell viability assay were performed to explore the impact of mi R-200 a on proliferation of HCT116 and SW480;Then Transwell migration and invasion experiments were used to probe the effect of mi R-200 a on colon cancer cell migration and invasion,and estimate the opposite effect of mi R-200 a and lncRNA H19 in CRC.To further study the role of mi R-200 a in colorectal cancer cells,the Tar Base v7.0 of DIANA TOOLS database were used to search possible target of mi R-200 a,and β-catenin was found to be suppressed by mi R-200 a.The Western blot experiment were performed to probe the relationship of mi R-200 a and its target gen.The Western blot experiment and TOP-Flash/FOP-Flash luciferase reporter assay were used to detect β-catenin expression and activity influenced by lncRNA H19.In order to verify the effects of lncRNA H19 on CRC,we measured lncRNA H19 expression in CRC tissues and adjacent tissues using quantitative real-time PCR(q RT-PCR),and compared lncRNA H19 expression levels with the dates of clinicopathologic characteristics of CRC patients.Results LncRNA H19 overexpression facilitated colon cancer cell proliferation,migration and invasion,whereas lncRNA H19 knockdown inhibited cell proliferation,migration and invasion.mi R-200 a bound to lncRNA H19 and inhibited its expression,thereby decreasing CRC cell proliferation,migration and invasion.β-catenin was identified as a target gene of mi R-200 a.LncRNA H19 up-regulatedβ-catenin expression and activity by competitively binding to mi R-200 a.LncRNA H19 expression was upregulated in CRC tissues compared with adjacent non-cancerous tissues,and increased with the tumor size and lymph node metastasis.Conclusion LncRNA H19 is up-regulated in CRC tissues and promotes cell proliferation,migration and invasion by competitively binding to mi R-200 a and derepressingβ-catenin in CRC.