The Study About Established Human Multidrug-resistant Osteosarcoma Cell Line MG63/ADR. And The Gene Expression Profiles Of Osteosarcoma Cells By Microarray Analysis

1,The study about established human multidrug-resistant osteosarcoma cell line MG63/ADR.Objective: In order lo reverse tumor MDR in vitro,we designed this study to establish human multidrug-resistant osteosarcoma cell line MG63/ADR.Methods: An adriamycin-resistant human osteosarcoma cell subline (MG63/ADR) was established in vitro using gradually increased concentration of adriamycin (ADM)in culture. CelI growth was measured and multidrug-resistance to multianticancer agents was evaluated by MTT assay. Flow cytometry(FCM) was performed to determine cell cycle.Western blot determine drug-resistance associated mdr,bcl-2,cyclinD1 and survivin gene. mRNA expression related to these proteins were determined with reverse transcription polymerase chain reaction (RT—PCR).Results: Compared to MG63,The number of MG63/ADR cells in G0/G1-phase was significantly decreased(10.61%) while those in S-phase increased 15.06%. Furthermore, MG63/ADR was resistant to many anti-tumor agents,and its IC50 of ADM was 9.09 times higher than that of parent cell line MG63. Significant overexpression of mdr,bcl-2,cyclinD1 and survivin protein were detecte. RT-PCR showed that mRNA expression of mdr,bcl-2,cyclinD1 and survivin were significantly increased in MG63/ADR cell.Conclusion: MG63/ADR is human multidrug-resistant,and its resistance may be closely related lo the overexpression of mdr,bcl-2,cyclinD1 and surviving. Objective: To study the gene expression profiles of osteosarcoma cells by microarray analysis.Methods:Total RNA was extracted from osteosarcoma MG63 and MG63/ADR cell. Illumina oligonucleotide microarrav was used to characterize 48 000 gene expression profile of each sample.Results: 1194 up-regulated genes and 1356 down-regulated genes were found in MG63/R cell compared to those in MG63 cell.Molecular and biological function analyses showed that the majority of the upregulated genes were associated with protein binding,translation control, transcriptional factors,metabolism,signal transduction,cell cycle,cell apoptosis,etal.Conclusion: These findings are expected to improve the understanding of the molecular mechanisms and the identification of therapeutic targets.