Establishment of multidrug resistant adenocarcinoma cell line A549/ADM model treated with serum free floating-culture system combined with doxorubicin

Objective: To establish a doxorubicin-resistant cell line A549/ADM and research the relationship between A549/ADM and human lung carcinoma stem cell.Methods: Doxorubicin-resistant lung adenocarcinoma cell line A549/ADM was obtained by treating lung adenocarcinoma A549 in vitro with increasing dosage of doxorubicin in serum free medium.The resistant to chemotherapeutic drugs was evaluated with MTT assay.Cell cycle,expressions of ABCB1 and ABCG2 and concentration of doxorubicin was determined by flow cytometry.CD133 positive cells explore were determined by the immunofluorescent staining and flow cytometry to examine lung carcinoma stem cell proportion.Tumorigenic potential was performed by nude mice xenograft transplant experiments.Results: A549/ADM cells were proved multidrug-resistant.The cells showed resistance to ADM,VCR,Taxol,and the resistant index of A549/ADM was 13.167,12.86,3.4.Cells in the period S was(45.76±1.41)% in A549 whereas(23.33±1.32)% in A549/ADM.The apoptosis of A549 cell was(58.23±0.82)% whereas(16.49±0.77)% in A549/ADM.Compared with the parental cell line,A549/ADM cell had a higher expression level of ABCB1 and ABCG2(P0.05).The concentration of doxorubicin in the A549/ADM is obviously lower than that in the A549 cell.The immunofluorescent staining and flow cytometry analysis showed that the expression levels of CD133 were significantly higher in A549/ADM cell.A549/ADM cell were highly tumorigenic in nude mice.Conclusion: Doxorubicin-resistant lung adenocarcinoma cell line A549/ADM was obtained by treating lung adenocarcinoma A549 in vitro with increasing dosage of doxorubicin in serum free medium.cell line A549/ADM had a higher proportion of lung adenocarcinoma stem cells compared to its parental cell line A549.