Characterization of a stem cell population in lung cancer A549 cells

Ji-Min Sung, Hee-Jung Cho, Hee Yi, Chi-Ho Lee, Hye-Sun Kim, Dong-Ku Kim, AM Adb El-Aty, Jin-Suk Kim, Christopher P. Landowski, Matthias Hediger, Ho-Chul Shin (Corresponding Author)

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Abstract

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.
Original languageEnglish
Pages (from-to)163-167
JournalBiochemical and Biophysical Research Communications
Volume371
Issue number1
DOIs
Publication statusPublished - 20 Jun 2008
MoE publication typeNot Eligible

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Stem cells
Methotrexate
Doxorubicin
Lung Neoplasms
Stem Cells
Cells
Side-Population Cells
Annexin A5
Population
Assays
Messenger RNA
A549 Cells
Multiple Drug Resistance
HOE 33342
tryptoquivaline
Regeneration
Cell Cycle
Staining and Labeling
Phenotype

Cite this

Sung, J-M., Cho, H-J., Yi, H., Lee, C-H., Kim, H-S., Kim, D-K., ... Shin, H-C. (2008). Characterization of a stem cell population in lung cancer A549 cells. Biochemical and Biophysical Research Communications, 371(1), 163-167. https://doi.org/10.1016/j.bbrc.2008.04.038
Sung, Ji-Min ; Cho, Hee-Jung ; Yi, Hee ; Lee, Chi-Ho ; Kim, Hye-Sun ; Kim, Dong-Ku ; Adb El-Aty, AM ; Kim, Jin-Suk ; Landowski, Christopher P. ; Hediger, Matthias ; Shin, Ho-Chul. / Characterization of a stem cell population in lung cancer A549 cells. In: Biochemical and Biophysical Research Communications. 2008 ; Vol. 371, No. 1. pp. 163-167.
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abstract = "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.",
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Sung, J-M, Cho, H-J, Yi, H, Lee, C-H, Kim, H-S, Kim, D-K, Adb El-Aty, AM, Kim, J-S, Landowski, CP, Hediger, M & Shin, H-C 2008, 'Characterization of a stem cell population in lung cancer A549 cells', Biochemical and Biophysical Research Communications, vol. 371, no. 1, pp. 163-167. https://doi.org/10.1016/j.bbrc.2008.04.038

Characterization of a stem cell population in lung cancer A549 cells. / Sung, Ji-Min; Cho, Hee-Jung; Yi, Hee; Lee, Chi-Ho; Kim, Hye-Sun; Kim, Dong-Ku; Adb El-Aty, AM; Kim, Jin-Suk; Landowski, Christopher P. ; Hediger, Matthias; Shin, Ho-Chul (Corresponding Author).

In: Biochemical and Biophysical Research Communications, Vol. 371, No. 1, 20.06.2008, p. 163-167.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Characterization of a stem cell population in lung cancer A549 cells

AU - Sung, Ji-Min

AU - Cho, Hee-Jung

AU - Yi, Hee

AU - Lee, Chi-Ho

AU - Kim, Hye-Sun

AU - Kim, Dong-Ku

AU - Adb El-Aty, AM

AU - Kim, Jin-Suk

AU - Landowski, Christopher P.

AU - Hediger, Matthias

AU - Shin, Ho-Chul

PY - 2008/6/20

Y1 - 2008/6/20

N2 - 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.

AB - 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.

U2 - 10.1016/j.bbrc.2008.04.038

DO - 10.1016/j.bbrc.2008.04.038

M3 - Article

VL - 371

SP - 163

EP - 167

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -