Influence of cyclophosphamide and its metabolites on PTEN gene after acting on ovarian cancer cell line SKOV3 / 重庆医学

ABSTRACT

Objective To study the effect of cyclophosphamide(CP) and its metabolites acrolein(ACR) on PTEN gene deleted on chromosome 10 after acting on ovarian cancer cellsSKOV3.Methods Different concentrations of CP and ACR were selected to act on recombinant PTEN protein.The phosphorylation activity of PTEN was detected by PNPP.The expression of PTEN protein was detected by Western blot.The binding mode of drug with protein was detected by the biotin combined with protein;meanwhile the expression change of P53/TP53 in PTEN gene pathway was analyzed.The target protein was obtained by immunoprecipitation(IP) after different drug concentrations acting on the cells.The phosphorylation activity of the target protein was detected by high performance liquid chromatography(HPLC).Results After the drug metabolites acting on recombinant PTEN protein,the phosphorylation activity was decreased with the increase of drug concentration,while the expression of ACR antibody action was increased with the drug concentration elevation.The expression of protein and biotin in different experimental groups was increased with the increase of drug concentration.The PTEN phosphorylation activity was decreased with the drug concentration increased in cells,and so did the expression of TP53 protein.Conclusion CP metabolite ACR induces the cytotoxicity by inhibiting PTEN protein phosphorylation activity.