The Impact Of Immune Suppressors On Liver Cancer Cell Line SK-hep-1Growth Is Medicated By Yap Gene

1Background and Specific AimsImmunosuppressive therapy is standard procedure against graft rejection afterliver transplantation. However, it is not well documented whether immunosuppressivetherapy impacts on liver cancer development, the mechanism underlying is unclear.Recent reports revealed that Yap gene may involves in liver cancer cell cycle, alongwith our initial finding, we hypothesis that Yap gene may involve in the impact ofimmune suppressor on liver cancer development. In current studies, liver cancerSK-hep-1cell line was utilized as cell model, tacrolimus, cyclosporine A, andrampamycin were used as immune suppressors, MTT, Cell flow cytometry,Yap-siRNA gene silence, Western-blot, Real-time PCR techniques were applied, wetry to study the impact of Yap gene on liver cancer cell development via immunesuppressors. The current studies will mainly focus on how we select immunesuppressors against both graft rejection and liver cancer development, we believe ourstudies may offer therapeutically stratagem initially.2ObjectsHuman Liver cancer cell line SK-hep-1, Ordered from American Type culturecollection (ATCC). 3Methods3.1The impact of immune suppressors on SK-hep-1was studied; MTT methodand flow cytometry was applied under three immune suppressors (FK5061000ng/ml,cyclosporine A and sirolimus100ng/ml) in SK-hep-1, respectively.3.2Yap gene was silenced by its specific siRNA, multiple techniques wereapplied, we studied the role of yap gene in liver cancer cell development.3.3Yap gene knock-down combined with immune suppressors treatment wasapplied, cancer cell cycle and immune factor genes were studied by its appropriatetechniques, we evaluated the impact of immune suppressors on SK-hep-1growthdevelopment. Statistic analysis was performed by Student’ t-test, p value＜0.05wasconsidered as statistical significance.4Results4.1FK5061000ng/ml and CsA600ng/ml trigger liver cancer cell lines SK-hep-1growth; however, Rapamycin100ng/ml inhibits SK-hep-1cell proliferation.4.2Yap gene knock-down experiments illustrate that yap gene promotesSK-hep-1cell growth.4.3Yap siRNA suppresses immune factor genes expression (IL2, B7H1,GM-CSF-1and IL8), it illustrates that yap gene impacts on the regulation of cellimmune factor.4.4Yap siRNA combined with FK506treatment in SK-hep-1, we illustrate thatYap gene might not involve in the promotion of SK-hep-1growth mediated byFK506.4.5Yap gene upregulates the SK-hep-1growth mediated by CsA treatment.4.6Yap gene attenuates RAP inhibition of SK-hep-1growth.5ConclusionImmune suppressors impact on liver cancer cell SK-hep-1differently.Tacrolimus and cyclosporine A trigger SK-hep-1growth, while sirolimus inhibits. Yapgene impacts the effect of SK-hep-1growth differently during immune suppressorstreatment, Yap gene promotes SK-hep-1growth mediated by CsA, but attenuates RAPinhibition of SK-hep-1growth, while it does not impact on SK-hep-1growthmedicated by FK506.