Open Access

Effects of 2-methoxyestradiol on apoptosis and HIF-1α and HIF‑2α expression in lung cancer cells under normoxia and hypoxia

  • Authors:
    • Arnoldo Aquino-Gálvez
    • Georgina González-Ávila
    • Javier Delgado-Tello
    • Manuel Castillejos-López
    • Criselda Mendoza-Milla
    • Joaquín Zúñiga
    • Marco Checa
    • Héctor Aquiles Maldonado-Martínez
    • Axel Trinidad-López
    • José Cisneros
    • Luz María Torres-Espíndola
    • Claudia Hernández-Jiménez
    • Bettina Sommer
    • Carlos Cabello-Gutiérrez
    • Luis H. Gutiérrez-González
  • View Affiliations

  • Published online on: November 4, 2015     https://doi.org/10.3892/or.2015.4399
  • Pages: 577-583
  • Copyright: © Aquino-Gálvez et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hypoxic tumor cells are known to be more resistant to conventional chemotherapy and radiation than normoxic cells. However, the effects of 2-methoxyestradiol (2-ME), an anti-angiogenic, antiproliferative and pro-apoptotic drug, on hypoxic lung cancer cells are unknown. The aim of the present study was to compare the effects of 2-ME on cell growth, apoptosis, hypoxia-inducible factor 1α (HIF-1α) and HIF-2α gene and protein expression in A549 cells under normoxic and hypoxic conditions. To establish the optimal 2-ME concentration with which to carry out the apoptosis assay and to examine mRNA and protein expression of HIFs, cell growth analysis was carried out through N-hexa-methylpararosaniline staining assays in A549 cell cultures treated with one of five different 2-ME concentrations at different times under normoxic or hypoxic growth conditions. The 2-ME concentration of 10 mM at 72 h was selected to perform all further experiments. Apoptotic cells were analyzed by flow cytometry. Western blotting was used to determine HIF-1α and HIF-2α protein expression in total cell extracts. Cellular localization of HIF-1α and HIF-2α was assessed by immunocytochemistry. HIF-1α and HIF-2α gene expression was determined by real‑time PCR. A significant increase in the percentage of apoptosis was observed when cells were treated with 2-ME under a normoxic but not under hypoxic conditions (p=0.006). HIF-1α and HIF-2α protein expression levels were significantly decreased in cells cultured under hypoxic conditions and treated with 2-ME (p<0.001). Furthermore, 2-ME decreased the HIF-1α and HIF-2α nuclear staining in cells cultured under hypoxia. The HIF-1α and HIF-2α mRNA levels were significantly lower when cells were exposed to 2-ME under normoxia and hypoxia. Our results suggest that 2-ME could have beneficial results when used with conventional chemotherapy in an attempt to lower the invasive and metastatic processes during cancer development due to its effects on the gene expression and protein synthesis of HIFs.
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January-2016
Volume 35 Issue 1

Print ISSN: 1021-335X
Online ISSN:1791-2431

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Spandidos Publications style
Aquino-Gálvez A, González-Ávila G, Delgado-Tello J, Castillejos-López M, Mendoza-Milla C, Zúñiga J, Checa M, Maldonado-Martínez HA, Trinidad-López A, Cisneros J, Cisneros J, et al: Effects of 2-methoxyestradiol on apoptosis and HIF-1α and HIF‑2α expression in lung cancer cells under normoxia and hypoxia. Oncol Rep 35: 577-583, 2016.
APA
Aquino-Gálvez, A., González-Ávila, G., Delgado-Tello, J., Castillejos-López, M., Mendoza-Milla, C., Zúñiga, J. ... Gutiérrez-González, L.H. (2016). Effects of 2-methoxyestradiol on apoptosis and HIF-1α and HIF‑2α expression in lung cancer cells under normoxia and hypoxia. Oncology Reports, 35, 577-583. https://doi.org/10.3892/or.2015.4399
MLA
Aquino-Gálvez, A., González-Ávila, G., Delgado-Tello, J., Castillejos-López, M., Mendoza-Milla, C., Zúñiga, J., Checa, M., Maldonado-Martínez, H. A., Trinidad-López, A., Cisneros, J., Torres-Espíndola, L. M., Hernández-Jiménez, C., Sommer, B., Cabello-Gutiérrez, C., Gutiérrez-González, L. H."Effects of 2-methoxyestradiol on apoptosis and HIF-1α and HIF‑2α expression in lung cancer cells under normoxia and hypoxia". Oncology Reports 35.1 (2016): 577-583.
Chicago
Aquino-Gálvez, A., González-Ávila, G., Delgado-Tello, J., Castillejos-López, M., Mendoza-Milla, C., Zúñiga, J., Checa, M., Maldonado-Martínez, H. A., Trinidad-López, A., Cisneros, J., Torres-Espíndola, L. M., Hernández-Jiménez, C., Sommer, B., Cabello-Gutiérrez, C., Gutiérrez-González, L. H."Effects of 2-methoxyestradiol on apoptosis and HIF-1α and HIF‑2α expression in lung cancer cells under normoxia and hypoxia". Oncology Reports 35, no. 1 (2016): 577-583. https://doi.org/10.3892/or.2015.4399