Establishment of new multidrug-resistant human osteosarcoma cell lines

Oncol Rep. Jul-Aug 2000;7(4):859-66. doi: 10.3892/or.7.4.859.

Abstract

Multidrug-resistant clones of human osteosarcoma MNNG/HOS and MG63 cells were isolated by stepwise selection on exposure to increasing doses of doxorubicin (DXR). The final clones MNNG/HOS/DXR1000 and MG63/DXR1000, established after ethylmethane sulfonate mutagenesis, showed 96-fold and 121-fold higer resistance to DXR than their parental cell lines. They were also cross-resistant to vincristine, but not to cisplatinum or methotrexate. The levels of multidrug-resistance-1 (MDR1) mRNA expression increased gradually according to the concentration of DXR in both cell lines. Although the parental MNNG/HOS cells expressed a low level of MDR1 mRNA, the parental MG63 cells showed no MDR1 expression. The IC50 values of MNNG/HOS and its resistant variant to DXR were higher than those of MG63 and its resistant clone. Multidrug-resistant associated protein (MRP) mRNA expression was detected in MNNG/HOS or MG63 parental cell lines, and in their resistant variants. MG63 and its resistant variants revealed stable expression of MRP, whereas the resistant phenotype of MNNG/HOS showed decreased MRP expression, compared to its parental cell line. No alteration in the levels of hepatocyte growth factor (HGF) or its receptor c-MET was recognized between parental lines and their resistant variants. The results indicate that our DXR-resistant variants of MNNG/HOS and MG63 reveal a classical MDR phenotype and can offer a model with which to investigate the mechanisms of multidrug resistance in osteosarcoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B, Member 1 / genetics*
  • Antineoplastic Agents / toxicity*
  • Bone Neoplasms / genetics
  • Bone Neoplasms / pathology*
  • Cell Culture Techniques / methods
  • Cell Survival / drug effects
  • Doxorubicin / toxicity
  • Drug Resistance, Multiple*
  • Humans
  • Methotrexate / toxicity
  • Methylnitronitrosoguanidine / toxicity
  • Osteosarcoma / genetics
  • Osteosarcoma / pathology*
  • Polymerase Chain Reaction
  • RNA, Messenger / genetics
  • Transcription, Genetic
  • Tumor Cells, Cultured
  • Vincristine / toxicity

Substances

  • ATP Binding Cassette Transporter, Subfamily B, Member 1
  • Antineoplastic Agents
  • RNA, Messenger
  • Methylnitronitrosoguanidine
  • Vincristine
  • Doxorubicin
  • Methotrexate