Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (37): 6913-6918.doi: 10.3969/j.issn.1673-8225.2011.37.018

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Establishment of adriamycin-resistant human osteosarcoma cell line and research on its biological characteristics

Chen Yu1, Wang Da-yong1, Weng Zheng1, Deng Zhong-liang2   

  1. 1Department of Orthopedics, Ninth People’s Hospital of Chongqing, Chongqing  400700, China
    2Department of Orthopedics, Second Affiliated Hospital of Chongqing Medical University, Chongqing  40010, China
  • Received:2011-06-10 Revised:2011-07-30 Online:2011-09-10 Published:2011-09-10
  • Contact: Deng Zhong-liang, Doctor, Professor, Department of Orthopedics, Second Affiliated Hospital of Chongqing Medical University, Chongqing 40010, China deng7586@gmail. com
  • About author:Chen Yu☆, Doctor, Attending physician, Department of Orthopedics, Ninth People’s Hospital of Chongqing, Chongqing 400700, China chenyu@vip.163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30772211*

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Abstract:

BACKGROUND: Multidrug resistance is a major factor leading to the failure of chemotherapy for human osteosarcoma. However, the precise mechanism remains poorly understood.
OBJECTIVE: To establish adriamycin-resistant human osteosarcoma cell line 143B/ADM and to analyze its biological characteristics.
METHODS: Increasing concentrations of adriamycin (ADM) were applied for 45 days to establish 143B/ADM resistant strain. The half of inhibiting concentrations (IC50) and resistance indexs (RI) of different antitumor drugs were measured by CCK assay, and the cell cycle was analyzed by flow cytometry. The cellular efflux capacity was estimated by rhodamine test. After the cell lines were treated by ADM, intracellular ADM concentration was detected by fluorospectrophotometer, and the apoptosis was observed by laser scanning confocal microscope and flow cytometry. Meanwhile, the expression of multidrug resistance 1 (MDR-1), multidrug resistance-associated protein 1 (MRP-1) and lung resistance protein (LRP) were detected by western blot analysis.
RESULTS AND CONCLUSION: After induction for 45 days, the RI of 143B/ADM cells to ADM was 15.7, and the 143B/ADM cells showed various resistances to cisplatin, methotrexate, isophosphamide, vincristine and taxinol. Compared with 143B/WT cell line, there were less cells in G2/M phase and more cells in G1 and S phases, markedly decreased intracellular rho123 and ADM in 143B/ADM cell line (P < 0.01). Flow cytometry and confocal laser microscopy analysis showed that at 72 hours after treatment with ADM (10 mg/L), cell apoptosis in 143B/ADM cells was less than that in 143B/WT cells. Furthermore, compared with 143B/WT, the MDR-1 expression of 143B/ADM was increased significantly with no difference of MRP-1 and LRP expression among both cell lines. Multidrug resistance of 143B/ADM cells is related to MDR-1, and has no relationship with MRP-1 and LRP. 

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