Effect Of CNOT7 Gene Knockdown On The Immune Microenvironment By Reducing The Secretion Of TGF-β In HepG2 Cells

Objective:To study the effect of human CCR4-NOT transcription complex subunit7(CN OT7)gene knockdown on the immune microenvironment of HepG2 cells.Methods:We designed a cell transfection protocol,and performed the experimentwith t hree groups:CNOT7-targeted knockdown group,control group,and CNOT7-over expression group.The transfection efficiency was assessed using inverted fluorescen ce microscopy.The expression level of CNOT7,transforming growth factor β(TGF-β),an d nuclear factor-kappa B(NF-κB)p65 proteins was determined by Western blotti ng.The concentration of TGF-β secreted in the cell culture supernatant was meas uredby ELISA.The sensitivity of tumor cells to the killing function of natural killer(NK)c ells wasdetected by flow cytometry.The effects of three cell culture supernatants on the secretion of human perip heral blood T cells was detected flow cytometry.Results:Compared with control group,the expression level of TGF-β and NF-κBp65proteins was significantly decreased in the CNOT7-targeted knockdown group,and the TGF-β concentration in the culture supernatant was also significantly reduced.However,in CNOT7 overexpression group,the expression level of two proteins a nd TGF-β concentration were significantly increased.NK cells were co-cultured wit h tumor cells,and the apoptosis rate of HepG2 cells transfected with CNOT7-spe cific shRNA was significantly increased.However,in CNOT7 overexpression group,the apoptosis rate was significantly decreased.The ability of T cells secreted by C NOT7-targeted knockdown group cell culture supernatants to secrete INF-γ and TNF-α was enhanced.Conclusion:CNOT7 forms the immune microenvironment of hepatocellular carcinoma.Targ eted knockdown of CNOT7 can reduce TGF-β secretion and enhance killing funct ion of NK cells toward HepG2 cells.Targeted knockdown of CNOT7 can reduce TGF-β levels and improve T cell secretion.CNOT7 gene knockdown can improve the immune microenvironment of hepatocellular carcinoma by reducing TGF-β secr etion in HepG2 cells.