Mechanism Study Of METTL5 Regulating TGFβ2 To Affect Proliferation And Migration In Hepatocellular Carcinoma Cells

Objective:To analyze the relationship between the expression of methyltransferase 5(RRNA N6-Adenosine-Methyltransferase METTL5)in the TGCA and HCCDB databases and survival prognosis,clinical stage and tumor grade by bioinformatics technology.To verify the expression of METTL5 by using fresh liver cancer tissue from hepatocellular carcinoma(HCC)patients and liver cancer cells.To clarify the effects of METTL5 expression changes on the proliferation and migration of hepatoma cells was determined by the function test in vitro.Then to explore the mechanism of METTL5 regulating the expression of Transforming growth factor-beta2(TGFβ2)on proliferation and migration of HCC cell.Methods:1.The relationship between the expression of METTL5 in HCC and survival prognosis,clinical stage and tumor grade of HCC patients were analyzed by using the public database in TGCA and HCCDB;2.The m RNA expression of METTL5 was detected using Real time PCR in liver cancer tissues,and the protein expression of METTL5 was detected using Western blot and immunohistochemistry(IHC);In cell lines,the m RNA expression level of METTL5 was detected using Real-time PCR and the protein expression level of METTL5 was detected using Western blot in human normal liver cells HL7702 and hepatoma cell lines HCCLM3,SMMC7721,Hep3 B,and MHCC97 H,respectively;3.The ability of cell proliferation changed by interferencing or overexpressing the expression of METTL5 were detected by CCK-8、EDU assay,The ability of cell migration changed by interferencing or overexpressing the expression of METTL5 were detected by wound assay、Transwell assay;4.Using HCC date in TCGA database to predict the possible related pathway or function of METTL5 in liver cancer,The correlation between the expression of METTL5 and TGFβ2 in hepatocellular carcinoma was analyzed in TCGA database;The changes of TGFβ2 m RNA and protein expression after the change of METTL5 expression in hepatoma cells were observed by Quantitative Real-time PCR and Western blotting;5.TGFβ2 was overexpressed in HCCLM3,a liver cancer cell line that stably interferes with METTL5 expression,and TGFβ2 was interfered in Hep3 B,a liver cancer cell line that overexpresses METTL5.then observed the protein expression level of METTL5 and TGFβ2 and the changes of cell proliferation and migration capacity.Results:1.To analysis TGCA database and HCCDB database showed that the expression of METTL5 in HCC was noticeably higher than that in normal liver tissue,and the high expression of METTL5 was an independent risk factor for poor prognosis of liver cancer patients;2.The expression of METTL5 in HCC tissues was high,and the expression of METTL5 in adjacent tissues was noticeably lower than it in HCC,and the expression of METTL5 in HCC cells was higher than it in normal liver cell;3.After interfering with METTL5,the capacity of proliferation and migration in hepatoma cells decreased。4.KEGG enrichment analysis suggests that METTL5 may through KEGG＿CELL＿CYCLE,KEGG＿NOTCH＿SIGNALING＿PATHWAY,KEGG＿P53＿SIGNALING＿PATHWAY,KEGG＿PATHWAYS＿IN＿CANCER,KEGG＿TGF＿BETA＿SIGNALING＿PATHWAY and KEGG＿WNT＿SIGNALING＿PATHWAY and other pathways to affect the progression of liver cancer,and correlation analysis suggests that METTL5 and TGFβ2 are closely related in liver cancer.5.After interfering METTL5 in hepatoma cells,the m RNA and protein expression levels of TGFβ2 were also down-regulated;TGFβ2 is a key effector molecule of METTL5 in regulating the proliferation and migration of hepatoma cells.Conclusion:METTL5 is up-regulated in liver cancer tissue and liver cancer cells and is associated with poor prognosis;interfering METTL5 can suppression the expression of TGFβ2,thus weakening the proliferation and migration of hepatoma cells;METTL5 is expected to become a new molecular marker for the treatment of liver cancer and provide treatment strategies for the diagnosis and treatment of liver cancer.