Activation Of Mice HCC Cell Proliferation By Hepatic Stellate Cells Though AKT Signaling

Background and aims Hepatocellular carcinoma (HCC) is known as a treatment-resistant malignancy which rising incidence in many regions on a global scale.lt is believed that HCC has a close connection with tumor microenvironment. Suppressing tumor progress through changing the tumor microenvironment provided a new approach of thinking for treament of HCC.The activation of hepatic stellate cells(HSCs) is an important part of the progression of hepatocarcinogenesis,at the same time,it also is regarded as an significant element of microenvironment in liver tumor.Now we aimed at analyzing the effects and mechanism of activated HSCs on the HCC cells proliferation both in vitro and in vivo.Methods In this study, to establish an orthotopic HCC and HSCs co-transplant model in mice, H22cells or H22cells plus HSCs were injected intrahepatically in BALB/c mice. After10days,we sacrificed the mice and took tumor samples, protein was isolated from tumor samples and detected by western blot for PERK/ERK and PI3K/AKT pathway gap-associated proteins’expression.What’s more,we used HSCs-CM (conditioned media),treated H22cells with CM and PI3K/AKT pathway specific inhibitor LY294002in vitro to investigate the effect of HSCs on H22cells and the mechanism between them. We put NIH3T3cells as negative control cells.At last,we injected H22cells which were treated by HSC-CM and NIH3T3-CM into BALB/c mice’s under the skin of oxter.Results The experiment successfully build orthotopic liver tumour and HSCs co-transplantation model in mice; The expression of p-AKT and its downstream protein s were significantly increased in the tumor sample of H22and HSCs co-transplanted mice. The expression of p-AKT and its downstream protein β-catenin-Cyclin D2and GSK3β-Cyclin D1in HSC-CM treated H22cells was significantly higher than the control group.Our date demonstrated that LY294002decreased cell proliferation both in control group and CM group.As well as the protein expression of p-AKT and its downsream protein. Conclusions,The results of our current study clearly suggest that cytokines secred by HSCs can obviously activate the PI3K/AKT signaling pathway both in vivo and in vitro.Our finding demonstrate that HSCs can promote H22cell proliferation through AKT signaling pathway and this ability is relatively independent,which provides a new way of thinking for the treatment of liver cancer.