Establishment of multidrug resistant cell line U251/VP16 of glioma and its biological characteristics

Abstract

Objective The multidrug resistant cell line U251/VP16 was established and induced by etoposide.Methods The U251 cells resistant to etoposide were make by increasing concentrations.The multidrug resistance was detected by cell counting Kit-8 (CCK-8).The morphology of U251 and U251/VP16 cells was observed after Wright-Giemsa staining.Cell cycle was tested by flow cytometry.Cell doubling time was calculated.The expression of multidrug resistance ( MDR1 ),B lymphocytes/leukemia-2(bcl-2),multidrug resistance associated protein 5 (MRP5) and low density lipoprotein receptor related protein 1 (LRP1) was assayed by using reverse transcription-polymerase chain reaction (RT-PCR).Results (1) Multidrug resistant cell line was constructed and the cell line had resistance to cyclophosphamide,etoposide,vincristine sulfate and doxorubicin hydrochloride; (2) The volume of multidrug resistance cell line was increased,showing pleomorphic under the optical microscope.Increased markedly irregular nucleus was observed and particles like substances were increased ; ( 3 ) The results of cell cycle showed that the proportion of U251/VP16 cells in G0/G1 and S phases was decreased,while that in G2/M phase was increased significantly; (4) The doubling time of U251/VP16 cells was (20.64 ± 1.98) h and that of U251 cells was (10.26 ± 1.03) h; (5) The expression of MDR1,bcl-2,MRP5 and LRP(1) was positive.Conclusion The multidrug resistance cell line was established and its drug resistance was stable.The multidrug resistance may be associated with bcl-2,MRP5,LRP1. Key words: Glioma; Etoposide; Multidrug resistance