Experimental Study On KMT2B Promoting Proliferation,Migration And Invasion Of Hepatocellular Carcinoma Cells By Up-regulating SNHG12

Objective: The purpose of this study is to clarify the expression level of KMT2B(histone lysine methyltransferase 2B)protein in human hepatocellular carcinoma(HCC)and liver cancer cells,and promote liver cancer cell invasion and migration of preliminary inquiry influence and potential molecular mechanisms.The impact and potential molecular mechanism may be related to its up-regulation of long non-coding RNA(lnc RNA)SNHG12 to recruit E2F1,which provides a new molecular target for the comprehensive treatment of hepatocellular carcinoma.Methods:Use TCGA database,GEPIA and Xiantao Academic to query the expression of KMT2 B protein in hepatocellular carcinoma,survival and prognosis;Using immunohistochemical staining experiment and q RT-PCR experiment to detect the expression level of KMT2 B in liver cancer tissues and adjacent normal tissues of 14 patients with primary liver cancer;based on bioinformatics analysis method using R software and multiple online data The platform conducts statistical analysis and visualization of data related to hepatocellular carcinoma in the TCGA database,studies and analyzes the expression of SNHG12 and E2F1 in hepatocellular carcinoma,survival and prognosis,and the correlation between KMT2 B protein and SNHG12 and E2F1;Western blot(WB)Detection of KMT2 B protein expression in human liver cancer cell lines Hu H7,BEL-7404,Hep-3B,Hep-G2 cells and normal liver cells HL-7702cells;construction of KMT2 B lentivirus knockdown cell model and SNHG12 plasmid knockdown,Overexpression cell model,observe the cell transfection by fluorescence microscope and use puromycin or geneticin to screen,and then check the KMT2 B protein and corresponding m RNA expression changes by Western blot and q RT-PCR to verify whether the infection is successful;use CCK8 cell proliferation Counting experiments,cell wound healing inspections,and Transwell chamber migration and invasion researchs were used to study the possible effects of knocking down KMT2 B,overexpressing SNHG12,and knocking down SNHG12 on the multiplication,migration,and invasion of liver cancer cell lines Hu H7 cells and BEL-7404 cells;construct KMT2 B After knocking down the lentivirus and then overexpressing the SNHG12 plasmid cell model,the expression of KMT2 B,SNHG12,and E2F1 in each cell model was detected by q RT-PCR experiments,and the influence and potential of KMT2 B protein in the multiplication,migration,and invasion of liver cancer cells were initially explored.molecular mechanism.Results: GEPIA showed that KMT2 B protein was highly expressed in human hepatocellular carcinoma tissues relative to paracancerous tissues,and the results were the same as immunohistochemical staining and q RT-PCR detection.In addition,KMT2 B was highly expressed in hepatocellular carcinoma cell lines Hu H7,BEL-7404,Hep-3B and Hep-G2 compared with normal hepatocytes HL-7702.The study based on bioinformatics analysis showed that SNHG12 and E2F1 were highly expressed in hepatocellular carcinoma and there might be a correlation between KMT2 B,SNHG12 and E2F1.The expression of KMT2 B,SNHG12 and E2F1 decreased significantly after lentivirus knockdown of KMT2 B gene,and inhibited the ability of proliferation,invasion and migration of hepatocellular carcinoma cells.After plasmid knockout and overexpression of SNHG12,the expression of E2F1 in hepatocellular carcinoma cells changed synchronously with SNHG12.Lentivirus knocks down KMT2 B and then plasmids overexpress SNHG12 and raised E2F1 expression,which promotes the multiplication and invasion of hepatocellular carcinoma cells.Conclusion: The evidence of this study shows that KMT2 B protein is highly expressed in live cancer tissues and hepatocellular carcinoma cells.KMT2 B protein may promote the proliferation,migration and invasion of hepatocellular carcinoma cells by raising the recruitment of E2F1 by SNHG12.