Effect Of AFP Inhibiting GATA5 Function Activation Of PI3K/AKT Pathway On Malignant Behavior Of Hepatocellular Carcinoma

Objective: To analyze the effect of GATA5 on the malignant behavior of cancer cell growth,migration and invasion after interference expression in liver cancer HLE cell line and GATA5 overexpression in liver cancer Hep G2 cell line;To analyze the regulation of GATA5 on PI3K/AKT pathway and the expression of oncogene Ras and Src;The effects of GATA5 on the regulation of Beta-catenin pathway and the expression of cancer stem cell factors C-myc and Sox2 were analyzed.Analyze the impact of AFP on GATA5 function.Methods: 1.Construction of cmv-GATA5 lentiviral expression vector;pc DNA3.1-AFP,p TT5-PTEN expression vector;construction of si RNA-GATA5,si RNA-AFP interference expression vector.2.MTT assay for the effect of GATA5 on the proliferation of hepatoma cells HLE and Hep G2.3.Transwell assay to detect cell migration.4.Cell scratch test to detect cell scratch repair ability.5.Western blot was used to detect the expression changes of the target protein.6.Immunofluorescence experiments,observation of target protein expression and positional changes by laser confocal microscopy.Results: cmv-GATA5 expresses GATA5 in Hep G2 cells,si RNA-GATA5 interferes with GATA5 expression in HLE cells,pc DNA3.1-AFP expresses AFP in HLE cells,si RNA-AFP interferes with AFP expression in Hep G2 cells,p TT5-PTEN is in HLE,PTEN is expressed in Hep G2 cells.GATA5 reduces the proliferation rate of cells and inhibits cell migration.GATA5 inhibits PI3K/AKT pathway and inhibits Ras and Src oncogene expression;GATA5 inhibits Beta-catenin pathway and inhibits stem cell factor Sox2 and C-myc expression;AFP inhibits GATA5 function and activates PI3K/AKT pathway.Conclusions: 1.GATA5 can inhibit the malignant behaviors such as proliferation,migration and scratch repair of HLE and Hep G2 cells.2.GATA5 can inhibit the AKT pathway and inhibit the expression of oncogene Ras and Src.GATA5 may inhibit the expression of stem cell factors C-myc and Sox2 by inhibiting the Beta-catenin pathway.3.AFP is able to inhibit GATA5 function activation of the PI3K/AKT pathway.