Effects Of Interleukin-17A On Stemness Of Hepatocellular Carcinoma Cell Lines

Objective To investigate the effects of IL-17A on sternness of hepatocellular carcinoma cells.Methods The supernatants of hepatocellular carcinoma cell lines Hep 33,HepG2,MHCC97H and MHCC97L were detected by ELISA.Then the cell lines were added into the soft fibrinogen and IL-17A was incorporated in the culture medium.The rate of sphere's formation and the sphere's diameters were measured after the cells were cultured for 7 days.Cells proliferation and migration were determined by Real Time Cellular Analysis(RTCA).The mRNA expressions of IL-17RA,IL-17RC and SOX2,NANOG,OCT4,BMI1,E-Cadherin,N-Cadherin,vimentin in cells were quantified by using SYBR Green real time polymerase chain reaction(qRT-PCR).The expression of E-cadherin,N-cadherin,vimentin were also detected by qRT-PCR and Western blot.Results Hepatocellular carcinoma cell lines Hep 3B,HepG2,MHCC97H and MHCC97L could not secrete interleukin-17A.The sphere's formation rate of Hep 3B cells increased significantly after the cells were cultured with 50ng/ml IL-17A for 7 days(t=5.533,P<0.001).The Hep 3B cells proliferation and migration were also enhanced in the presence of 50ng/ml IL-17A.In Hep 3B cells the mRNA expressions of IL-17RA,IL-17RC,SOX2,NANOG,OCT4 and BMII were significantly upregulated after the stimulation with IL-17A.E-cadherin,which related to epithelial to mesenchymal transition(EMT)was downregulated firstly and then upregulated after IL-17A treatment,N-cadherin was upregulated,vimentin was upregulated firstly and then downregulated.Conclusions IL-17A can enhance the sternness of some hepatoceullular carcinoma cells.