Establishment of a methotrexate-resistant cell line of human osteosarcoma and its biological characteristics
- Author:
Jian-Jun WANG
1
Author Information
1. Department of Medical Oncology
- Publication Type:Journal Article
- Keywords:
Cell, Saos-2;
Drug resistance, neoplasm;
Methotrexate;
Osteosarcoma
- From:
Tumor
2007;27(8):623-627
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To establish a methotrexate (MTX)-resistant human osteosarcoma cell line (Saos-2/MTX1 and Saos-2/MTX2) Lived from the Saos-2 cell line and observe its biological characters. Methods: Saos-2 cells were pulse-exposed to MTX with gradually increased dosage. Both MTX-resistant human osteosarcoma cell lines (Saos-2/MTX1 and Saos-2/MTX2) were established after 178 days. The sensitivity of the Saos-2, Saos-2/MTX1, and Saos-2/MTX2 cell lines to MTX, cisplatin (DDP), doxorubicin (ADM), ifosfamide (IFO), epirubicine (EPI), pirarubicin (PTH), and paclitaxel (PTX) was measured by MTT assay. The morphological and ultra-structural changes of the cells were observed by optical microscopy and transmission electronic microscopy. The cell proliferation ability was calculated from the growth curve. The cell cycle distribution was analyzed by flow cytometry. Results: The MTX-resistant human osteosarcoma cell lines, Saos-2/MTX1 and Saos-2/MTX2, were established after 178 d induction. The resistance indexes to MTX were 4.87 and 12.73 times than that of Saos-2, respectively. The two cell lines had cross resistance to ADM, EPI, PTH and TAXOL and were still sensitive to DDP (P > 0.05). Optical microscopy showed the volume of Saos-2/MTX1 and Saos-2/MTX2 cells were enlarged and the number of poly-nucleus cells was increased. Transmission electronic microscopy revealed the cell process was decreased and the nucleoli were increased. Cell proliferation curve and flow cytometry demonstrated that the proliferation abilities of Saos-2/MTX 1 and Saos-2/MTX 2 cells were decreased significantly and the proportion of cells in S phase was reduced (P < 0.05). Conclusion: We establish two MTX-resistant human osteosarcoma cell lines (Saos-2/MTX1 and Saos-2/MTX2) and observe their biological characteristics. It provides a new experimental model for further studying the mechanism of MTX-resistance.
