Characterization of a stem cell population in lung cancer A549 cells

• Published in: Biochemical and biophysical research communications Vol. 371; no. 1; pp. 163 - 167
• Main Authors: Sung, Ji-Min, Cho, Hee-Jung, Yi, Hee, Lee, Chi-Ho, Kim, Hye-Sun, Kim, Dong-Ku, Abd El-Aty, A.M., Kim, Jin-Suk, Landowski, Christopher P., Hediger, Matthias A., Shin, Ho-Chul
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20-06-2008
• Subjects: A549 cells; ABCG2; Antineoplastic Agents - pharmacology; Apoptosis; ATP Binding Cassette Transporter, Sub-Family G, Member 2; ATP-Binding Cassette Transporters - genetics; ATP-Binding Cassette Transporters - metabolism; Cancer stem cells; Cell Cycle - drug effects; Cell Line, Tumor; Cell Proliferation - drug effects; Doxorubicin - pharmacology; Doxorubicin/methotrexate; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Gene Expression; Humans; Lung Neoplasms - metabolism; Lung Neoplasms - pathology; Membrane Transport Proteins - genetics; Membrane Transport Proteins - metabolism; Methotrexate - pharmacology; Multidrug Resistance-Associated Proteins - genetics; Multidrug Resistance-Associated Proteins - metabolism; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; Neoplastic Stem Cells - drug effects; Neoplastic Stem Cells - metabolism; Neoplastic Stem Cells - pathology; Side population; Doxorubicin/methotrexate; A549 cells; Cancer stem cells; ABCG2; Side population
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2008.04.038

We isolated a stem cell subpopulation from human lung cancer A549 cells using FACS/Hoechst 33342. This side population (SP), which comprised 24% of the total cell population, totally disappeared after treatment with the selective ABCG 2 inhibitor fumitremorgin C. In a repopulation study, isolated SP and non-SP cells were each able to generate a heterogeneous population of SP and non-SP cells, but this repopulation occurred more rapidly in SP cells than non-SP. An MTT assay and cell cycle distribution analysis reveal a similar profile between SP and non-SP groups. However, in the presence of doxorubicin (DOX) and methotrexate (MTX), SP cells showed significantly lower Annexin V staining when compared to non-SP cells. Taken together, these results demonstrate that SP cells have an active regeneration capacity and high anti-apoptotic activity compared with non-SP cells. Furthermore, our GeneChip
® data revealed a heightened mRNA expression of ABCG2 and ABCC2 in SP cells. Overall these data explain why the SP of A549 has a unique ability to resist DOX and MTX treatments. Therefore, we suggest that the expression of the ABCG2 transporter plays an important role in the multidrug resistance phenotype of A549 SP cells.