ROS production by endogenously generated protoporphyrin ix in murine leukemia cells

B Diez; R Cordo Russo; M J Teijo; S Hajos; A Batlle; H Fukuda

Issue

Vol. 55 No. 2: Porphyrias and associated pathologies. Biochemistry and molecular biology Part 2

Issue Published : July 1, 2009

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ROS production by endogenously generated protoporphyrin ix in murine leukemia cells

B Diez

Universidad de Buenos Aires Departamento de Quimica Biologica, FCEN Argentina

R Cordo Russo

Universidad de Buenos Aires Catedra de Inmunologia, IDEHU, Facultad de Farmacia y Bioquimica Argentina

M J Teijo

Universidad de Buenos Aires Departamento de Quimica Biologica, FCEN Argentina

S Hajos

Universidad de Buenos Aires Catedra de Inmunologia, IDEHU, Facultad de Farmacia y Bioquimica Argentina

A Batlle

Universidad de Buenos Aires Centro de Investigaciones sobre Porfirinas y Porfirias (CIPYP), CONICET Argentina

H Fukuda

Universidad de Buenos Aires Departamento de Quimica Biologica, FCEN Argentina

Corresponding Author(s) : B Diez

Cellular and Molecular Biology, Vol. 55 No. 2: Porphyrias and associated pathologies. Biochemistry and molecular biology Part 2
Article Published : July 1, 2009

Abstract

Endogenous production of Protoporphyrin IX (PpIX) is successfully exploited for photodynamic therapy (PDT) on malignant cells, following 5-aminolevulinic acid (ALA) administration and light irradiation. This treatment kills cancer cells by damaging organelles and impairing metabolic pathways via cellular reactive oxygen species (ROS) generation. We studied the efficiency of PpIX synthetized from ALA on ROS generation, in the Vincristine resistant (LBR-V160), Doxorubicin resistant (LBR-D160) and sensitive (LBR-) murine leukemia cell lines. Cells were incubated 4 hr with 1 mM ALA and then irradiated during different times with fluorescent light. One hour later, production of ROS was analyzed by flow cytometry using different fluorescent probes: Hydroethidine (HE) for superoxide anion, 2',7' Dichlorodihydrofluorescein diacetate (DCFH-DA) for hydrogen peroxide; mitochondrial damage was examined with 3,3' Dihexyloxacarbocyanine iodide (DiOC6). We found that superoxide anion production in the three cell lines increased with irradiation time whereas no peroxide hydrogen was detected. Mitochondrial damage also increased in an irradiation time dependent manner, being higher in the Vincristine resistant line. Previous studies have demonstrated that apoptotic cell death increased with irradiation time, which is consistent with these results, indicating that ROS are critical in ALA-PDT efficiency to kill malignant cells.

Keywords

5-aminolevulinic acid porphyrins photodynamic therapy.

Diez, B., Cordo Russo, R., Teijo, M. J., Hajos, S., Batlle, A., & Fukuda, H. (2009). ROS production by endogenously generated protoporphyrin ix in murine leukemia cells. Cellular and Molecular Biology, 55(2), 15–19. Retrieved from https://mail.cellmolbiol.org/index.php/CMB/article/view/1082