Abstract：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（P<0.05）.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.
[1] Clarke MF, Dick JE, Dirks PB, et al. Cancer stem cells perspectives on current status and future directions: AACR workshop on cancer stem cells[J]. Cancer Res, 2006, 66 (19):9339-9344.[2] Al Hajj M, Wicha MS, Benito Hernandez A, et al. Prospective identification of tumorigenic breast cancer cells[J]. Proc Natl Acad Sci USA,2003, 100(7): 3983-3988.[3] Dean M, Fojo T, Bates S. Tumour stem cells and drug resistance[J]. Nat Rev Cancer, 2005,5(4): 275-284.[4] Eramo A, Lotti F, Sette G, et al. Identification and expansion of the tumorigenic lung cancer stem cell population[J]. Cell Death Differ, 2008, 15(3): 504-514.[5] Reya T, Morrison SJ, Clarke MF, et al. Stem cells, cancer, and cancer stem cells[J]. Nature, 2001, 414(6859）: 105-111.[6] Yang LY, Trujillo JM. Biological characterization of multidrug resistant human colon carcinoma sublines induced/selected by two methods［J］. Cancer Res, 1990， 50（11）: 3218-3225.[7] Goodell MA, Brose K, Paradis G, et al. Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo[J]. J Exp Med, 1996, 183(4) : 1797-1806.[8] 易娟，陈静，孙静，等. 白血病K562/ADM细胞耐药性与白血病干细胞及耐药蛋白表达的关系[J].中华医学杂志，2009，89（25）：1741-1744.[9] 陈元文，吴诚义，陈鑫，等. 长程无血清培养人乳腺癌细胞球的生物学特性[J]. 肿瘤，2010，30(4):283-287.[10] 安勇，姚捷，卫积书，等. 吉西他滨耐药人胰腺癌细胞株的建立及其与肿瘤干细胞的相关性研究[J].中华外科杂志，2010，48（13）：999-1003.[11] 潘泓，谢彤，茅乃权，等. A549/DDP耐药肺癌细胞系的建立及其与SP细胞的关系[J].实用医学杂志，2010，26（14）：2477-2479.
2012, Vol. 22 
