Publicado

2018-10-01

Hypoxia-inducible factor HIF-1α modulates drugs resistance in colon cancer cells

Factor inducible por hipoxia HIF-1α modula la resistencia a drogas en células de cáncer de colon

Palabras clave:

Apoptosis, Cell Hypoxia, Colon Cancer, Doxorubicin (en)
Apoptosis, Hipoxia celular, Cáncer del colon, Doxorrubicina (es)

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Autores/as

  • Martha Leonor Pinzón-Daza Universidad del Rosario - School of Medicine and Health Sciences - Clinical Research Group - Bogotá D.C. - Colombia
  • Yenith Cuellar Universidad Nacional de Colombia - Bogotá Campus - Faculty of Science - Master’s Degree program in Biochemistry - Bogotá D.C. - Colombia
  • Alejandro Ondo Universidad del Rosario - School of Medicine and Health Sciences - Clinical Research Group - Bogotá D.C. - Colombia
  • Luisa Matheus Universidad del Rosario - School of Medicine and Health Sciences - Neuroscience Research Group - Bogotá D.C. - Colombia
  • Lilia Del Riesgo Universidad del Rosario - School of Medicine and Health Sciences - Neuroscience Research Group - Bogotá D.C. - Colombia
  • Fabio Castillo Universidad del Rosario - School of Medicine and Health Sciences - Doctoral program in Biomedical and Biological Sciences - Bogotá D.C. - Colombia
  • Ruth Garzón Universidad del Rosario - School of Natural Sciences and Mathematics - Biochemistry and Biotechnology Research Group (Bio Bio) - Bogotá D.C. - Colombia

Introduction: Drug resistance mechanisms may be associated with decreased cell death and its induction may depend on the response to oxidative stress caused by hypoxia. The correlation between hypoxia-inducible factor HIF-1α, the number of reactive oxygen species and their effect on cell survival has not yet been evaluated.

Objective: The purpose of this study was to evaluate the effect of HIF-1α activity and reactive oxygen species (ROS) accumulation in apoptosis of colon cancer cells.

Materials and methods: HT29 colon cancer cells were treated with CoCl2 or doxorubicin and the activity of HIF-1α was determined by ELISA assay. ROS were determined using fluorescence probe carboxy-H2DFFDA. Apoptosis was assessed by caspase-3 activation analysis, and PUMA and BAX mRNA levels by qRT-PCR. The reduction of the antiapoptotic effect due to hypoxia was attenuated by use of the endonuclease APE-1 (E3330) inhibitor. The endonuclease E3330 APE-1 inhibitor allowed evaluating the effect of ROS generated by doxorubicin and CoCl2 on apoptosis.

Results: Chemical hypoxia in combination with doxorubicin is an oxidative stressor in HT29 cells and induces a reduction in the apoptotic process in a time-dependent manner.

Conclusion: Resistance to hypoxia and doxorubicin-mediated cell death could be controlled by a mechanism related to the activity of HIF-1α and the amount of reactive oxygen species generated.

Introducción. Los mecanismos de resistencia a drogas podrían asociarse con disminución en la muerte celular y su inducción podría depender de la respuesta al estrés oxidativo que origina la hipoxia. La correlación entre factor inducible por hipoxia HIF-1α, cantidad de especies reactivas de oxígeno y su efecto sobre la supervivencia celular aún no ha sido evaluada.

Objetivo. Evaluar el efecto de la inducción de la actividad de HIF-1α y la cantidad de especies reactivas de oxígeno sobre la apoptosis en células de cáncer de colon.

Materiales y métodos. Células de cáncer de colon HT29 fueron tratadas con cloruro de cobalto (CoCl2) o doxorrubicina, luego se evaluó la actividad de HIF-1α por ELISA. Las especies reactivas de oxígeno fueron determinadas con sonda fluorescente carboxi-H2DFFDA. La apoptosis fue evaluada por la actividad de caspasa-3 y los niveles de mRNA de los genes proapoptóticos PUMA y BAX por qRT-PCR. El inhibidor de la endonucleasa APE-1 E3330 permitió evaluar el efecto de las especies reactivas de oxígeno generadas por doxorubicina y CoCl2 sobre la apoptosis.

Resultados. La hipoxia química combinada + doxorubicina es estresor oxidativos en células HT29 e induce una reducción en el proceso apoptótico de manera tiempo dependiente.

Conclusión. La resistencia a la muerte celular mediada por hipoxia y doxorubicina podría estar controlada por un mecanismo relacionado con la actividad de HIF-1α y la cantidad de especies reactivas de oxígeno generadas.


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