The difference of ADM binding protein in human breast cancer cell line MCF-7/Wild and MCF-7/ADM Cell / 中国药理学通报

ABSTRACT

Aim To study the mutidrugresistance mechanism of tumor cell by comparing the binding pro-tein with Adriamycin(ADM)in the MCF-7/Wild cell and MCF-7/ADM resistance cell.Methods The biotin-ADM compound was synthesized with the esterification-dehydration method,purified with HPLC and analyzed with mass spectrograph.The binding proteins against Adriamycin from MCF-7/ADM or MCF-7/Wild cell whose resistance index is above 200 were obtained respectively by the affinity screening magnetism bead method with biotin-ADM compound as a specific ligand.Some binding proteins with Adriamycin were identified with the MALDI-TOF-MS after isolating them with SDS-PAGE.Cell motor ability was compared in the migration experiment.Results The variability of biotin-ADM and ADM was negligible,while that IC50 of the two compounds to MCF-7/Wild was 0.796 ?mol?L-1 and 0.547 ?mol?L-1 respectively.The 20 types binding proteins with ADM were found from MCF-7/Wild cell,and 17 types binding proteins from MCF-7/ADM resistance cell.There was one protein band difference from the two cells on SDS-PAGE with silver staining.The same binding proteins on the two cells that identified with MALDI-TOF-MS were myosin,beta-actin,alpha-actin,keratin.MCF-7/W and MCF-7/ADM both migrated after being cultivated 72 h,with a faster migration of MCF-7/ADM.Conclusion There are different binding protein with ADM on the MCF-7/Wild cell and MCF-7/ADM resistance cell.The same binding protein with ADM on the two cells belongs to the cystoskeleton protein.The binding of the ADM with the cell cystoskeleton protein has some effect to the migration of the MCF-7/ADM.