Objective: To establish a multidrug-resistant human lung adenocarcinoma cell line and to investigate its biologic characteristics.
Methods: NBV of the terminal concentration of 0.02 mg/L was co-cultured with the human lung adenocarcinoma cells of the line Anip973. When the cells got a stable growth and generation the concentration of NVB was increased gradually till the 65 th generation. Thus a drug-resistant line Anip973/NVB that could grow under the NVB of the concentration of 2.0 mg/L was established. Anip973 and Anip973/NVB cells were co-cultured with 10 anti-cancer drugs: NVB, cisplatin, fluorouracil, gemcitabine, paclitaxel, etoposide, irinotecan, dacarbazine, ifosfamide, and pharmorubicin of different concentrations respectively. Forty-eight hours later MTT method was used to detect the 50% inhibition concentration (IC50) values of different drugs. The doubling times of the Anip973 and Anip973/NVB cells were calculated. Inverted microscopy and electron microscopy were used to observe the morphology of the cells. Flow cytometry was conducted to observe the cell cycle. The cells were cultured in the medium with NVB of the concentration of 2.0 mg/L. High efficiency fluid chromatography was used to observe the drug concentration in the cells.
Results: Anip973/NVB cells showed resistance at different degrees to 8 drugs (all P<0.05), except to gemcitabine and irinotecan. The doubling time of the Anip973 cells was 23.45 h, not significantly different from that of the Anip973/NVB cells The percentage of the cells in the phase G0 approximately G1 was 62.90% in the Anip973 cells, significantly higher than in the Anip973 cells (53.73%, P=0.014), and the proportion of the cells in the phase S 28.32% in the Anip973/NVB cells, significantly lower than that in the Anip793 cells (38.25%, P=0.035) NVB could be detected in the Anip973 cells with a concentration of 0.22 mg/L+/-0.05 mg/L, however, could not be detected in the Anip973/NVB cells. MTT assay showed that the IC50 of Anip973/NVB cells was 21.81 times higher than Anip973 cells. Microscopy showed that the structure, especially the ultramicrostructure, of the Anip973/NVB cells was more irregular than that of the Anip973 cells, and there were significant difference in ultramicrostructure.
Conclusion: A reliable multi-drug resistant human lung adenocarcinoma cell line Anip973/NVB has been successfully established.