Establishment of an adriamycin resistance cell line of human osteosarcoma and study of its resistant mechanisms / 中国药理学通报

ABSTRACT

Aim To establish a resistant human osteosarcoma cell line(Saos-2/ADM1and Saos-2/ADM4)from the Saos-2 cell line and study its resistant mechanisms.Methods Saos-2 cells were pulse exposed in gradually increased dose of ADM culturemedium.The sensitivity of the resistance drug from the Saos-2、Saos-2/ADM1 and Saos-2/ADM4 cell lines to ADM、DDP、EPI、MTX、THP and PTX was measured by MTT assay.The morphology and ultramicro structure of the cell lines were observed by optical microscopy and transmission electronic microscopy.The expression level of MDR1 mRNA、MRP mRNA and their proteins P-gp、MRP was examined by reverse transcription polymerase chain reaction and immunohistochemistry.Results Resistance cell lines were established after 167 days.The resistance index to methotrexate of the Saos-2/ADM1and Saos-2/ADM4 cell lines to ADM was 49.8 and 74.6 times than that of Saos-2 respectively.The two cell lines had resistance to MTX、IFO、EPI、THP and PTX(P0.05).Disordered structure of the Saos-2/ADM1and Saos-2/ADM4 cells was observed through microscopy.The cells appeared in coenocytic.The decrease of cell villus and the increase of nucleoli were observed through transmission electronic microscopy.The proliferation ability of Saos-2/ADM1and Saos-2/ADM4 cells decreased significantly.MDR1 mRNA、MRP mRNA、P-gp and MRP showed positive staining in resistance cell lines.Conclusion The genes of MDR1 mRNA、MRP mRNA and their corresponding proteins participated in the formation of multidrug resistance in resistant adriamycin cell line.These newly-described resistant osteosarcoma cell lines were useful models for further characterization of drug resistance in osteosarcoma and for the development of treatment protocol.