ABSTRACT
Objective To investigate induction of apoptosis of human ovarian cancer CoC1 cells by 5-allyl-7-gen-difluoromethylenechrysin(ADFMChR)in vitro,and its molecular mechanism. Methods The proliferative and growth inhibition of CoC1 cells treated with ADFMChR was respectively measured using(3,4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide(MTT)colorimetric assay and clone-formation in soft agar assay. The apoptosis of CoC1 cells induced by ADFMChR was determined by DNA agarose gel electrophoresis assay. Effect of ADFMChR on PPAR-γ, Bcl-2. Bax protein expression level of CoC1 cells was detected by western blotting. Results The proliferation of CoC1 cells could be significantly inhibited by ADFMChR in a dose-dependent manner. The IC50 was 7. 76 μmol/L The ladder-shaped band could be shown in DNA agarose gel electrophoresis after treatment with ADFMChR at 30. 0 μmol/L for 48 h and the ladder-shape band disappeared with GW9662. Western blot analysis showed that expression of PPAR-γ and Bax proteins were up-regulated and protein levels of Bcl-2 were depressed after treatment with ADFMChR in a concentration-dependent fashion. However,the effects of ADFMChR on Bcl-2 and Bax proteins were blocked or diminished in the presence of GW9662. Conclusions The effect of ADFMChR on induction of apoptosis in CoC1 cells may be mediated by activation of PPAR-γ, sequentially accompanied by reducing of protein levels of Bcl-2 and increasing of Bax expression.
ABSTRACT
Yeast surface display involves that the exogenous protein, which was fused with the yeast outer shell cell wall protein, was genetically anchored on the yeast cell surface. It has been widely used in expression and screening of functional protein. Here, we focused on the construction of lipase-displaying systems and its application in enzymatic biosynthesis, such as fatty acid methyl esters, short-chain flavour esters and sugar esters applications, and so on.