Preliminary Mechanism Study On The (20S)G-Rh2-induced Inhibitory Effect Of SK-HEP-1 Cell Migration

(20S)G-Rh2,a trace ingredient of ginseng,significantly inhibits tumor cell growth and triggers both mitochondrial cytochrome c release mediated endogenous apoptosis and Fas-mediated extrinsic apoptosis pathway to cause extensive tumor cell death.This thesis is aimed at identifying cellular target of(20S)G-Rh2 via biochemical and cellular biological method,and revealing underlying molecular mechanism.Liver cancer is known as a type of fatal malignancy with a second highest lethality rate throughout the world and China present an extremely higher level than the average.With a rapid development,most liver cancer cells exhibit superior migration and metastasis.So far,there is no effective treatment for liver cancer.Therefore,it is very important to find small molecule drugs that can effectively:inhibit the migration of liver cancer cell.The migration is a hallmark of cancer.Annexin A2 is abnormally overexpressed in most of cancers and plays multiple roles in the regulation of tumor cell physiology.AIIt,a heterotetramer formed by Annexin A2 and S100A10,is considered as a key player in cancer cell migration.In this thesis we carried out the following research:i)cell scratch healing assay showed that(20S)G-Rh2 significantly inhibited the migration of human hepatoma SK-HEP-1 cells;ii)we constructed prokaryotic expression plasmids of Annexin A2 and S100A10;iii)using E.coli expression system,we successfully expressed and purified Annexin A2 and S100A10 proteins;iv)we demonstrated the interaction between Annexin A2 and(20S)G-Rh2 by Isothermal titration calorimetry(ITC),and the binding constant is 1.13 × 105 ± 6.71 × 104M-1;v)Co-IP experiments demonstrated that(20S)G-Rh2 can inhibit the interaction of S100A10 and Annexin A2 effectively.Taken together,we suggest that(20S)G-Rh2 may intervened the formation of AIIt by interact with Annexin A2,and finally inhibit the migration of liver cancer cells.This thesis provided a preliminary molecular basis for further study on anti-cancer effect and mechanisms of(20S)G-Rh2.