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Delta-aminolevulinic acid cytotoxic effects on human hepatocarcinoma cell lines.

De Siervi A, Vazquez ES, Rezaval C, Rossetti MV, del Batlle AM - BMC Cancer (2002)

Bottom Line: Because apoptosis is usually associated with DNA fragmentation, the DNA of ALA treated and untreated cells were analyzed.To elucidate the mechanisms of ALA induced apoptosis, we examined its effect on p53 expression.CDK2 and CDK4 protein levels were reduced after ALA treatment at physiological concentrations.

View Article: PubMed Central - HTML - PubMed

Affiliation: Centro de Investigaciones sobre Porfirinas y Porfirias, Argentine National Research Council (CONICET), Department of Biological Chemistry, FCEN, University of Buenos Aires, Argentina. desiervi@qb.fcen.uba.ar

ABSTRACT

Background: Acute Intermittent Porphyria is a genetic disorder of heme metabolism, characterized by increased levels of porphyrin precursors, delta-aminolevulinic acid (ALA) and porphobilinogen (PBG). ALA has been reported to generate reactive oxygen species and to cause oxidative damage to proteins, subcellular structures and DNA. It is known that oxidative stress can induce apoptosis. The aim of this work was to study the cytotoxic effect of ALA on two hepatocarcinoma cell lines.

Results: We have determined the impact of ALA on HEP G2 and HEP 3B hepatocarcinoma cell lines survival as measured by the MTT assay. ALA proved to be cytotoxic in both cell lines however; HEP G2 was more sensitive to ALA than HEP 3B. Addition of hemin or glucose diminished ALA cytotoxicity in HEP G2 cells; instead it was enhanced in HEP 3B cells. Because apoptosis is usually associated with DNA fragmentation, the DNA of ALA treated and untreated cells were analyzed. The characteristic pattern of DNA fragmentation ladders was observed in ALA treated cells. To elucidate the mechanisms of ALA induced apoptosis, we examined its effect on p53 expression. No changes in p53 mRNA levels were observed after exposure of both cell lines to ALA for 24 h. CDK2 and CDK4 protein levels were reduced after ALA treatment at physiological concentrations.

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Cytotoxic effects of ALA. Human hepatocarcinoma cell lines (HEP G2 (circles) HEP 3B (squares)) were grown in the absence or in the presence of increasing ALA concentrations (0.5 to 5 mM) and the viability of cells was determined by the MTT assay. Viability was expressed in terms of population growth, as percent of an untreated control population with standard error of the mean (SEM).
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Figure 1: Cytotoxic effects of ALA. Human hepatocarcinoma cell lines (HEP G2 (circles) HEP 3B (squares)) were grown in the absence or in the presence of increasing ALA concentrations (0.5 to 5 mM) and the viability of cells was determined by the MTT assay. Viability was expressed in terms of population growth, as percent of an untreated control population with standard error of the mean (SEM).

Mentions: Human hepatocarcinoma cells were treated with various concentrations (0.5–5 mM) of ALA and the viability of cells was determined (Figure 1). ALA was toxic to both hepatocarcinoma cell lines. In HEP G2 cells, cytotoxicity occurred after exposure to 0.5 mM ALA for 24 h (growth inhibition: 30%) and the highest growth inhibition (70%) was observed with the highest ALA concentration (5 mM). HEP 3B cells were more resistant to ALA than HEP G2 cells, exhibiting only 12% growth inhibition after exposure to 0.5 mM ALA for 24 h and a 40% growth inhibition with 5 mM ALA. The strange shape of the viability curves obtained in both cases, with a restoration of control viability with 2 mM ALA, cannot be explained.


Delta-aminolevulinic acid cytotoxic effects on human hepatocarcinoma cell lines.

De Siervi A, Vazquez ES, Rezaval C, Rossetti MV, del Batlle AM - BMC Cancer (2002)

Cytotoxic effects of ALA. Human hepatocarcinoma cell lines (HEP G2 (circles) HEP 3B (squares)) were grown in the absence or in the presence of increasing ALA concentrations (0.5 to 5 mM) and the viability of cells was determined by the MTT assay. Viability was expressed in terms of population growth, as percent of an untreated control population with standard error of the mean (SEM).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC101407&req=5

Figure 1: Cytotoxic effects of ALA. Human hepatocarcinoma cell lines (HEP G2 (circles) HEP 3B (squares)) were grown in the absence or in the presence of increasing ALA concentrations (0.5 to 5 mM) and the viability of cells was determined by the MTT assay. Viability was expressed in terms of population growth, as percent of an untreated control population with standard error of the mean (SEM).
Mentions: Human hepatocarcinoma cells were treated with various concentrations (0.5–5 mM) of ALA and the viability of cells was determined (Figure 1). ALA was toxic to both hepatocarcinoma cell lines. In HEP G2 cells, cytotoxicity occurred after exposure to 0.5 mM ALA for 24 h (growth inhibition: 30%) and the highest growth inhibition (70%) was observed with the highest ALA concentration (5 mM). HEP 3B cells were more resistant to ALA than HEP G2 cells, exhibiting only 12% growth inhibition after exposure to 0.5 mM ALA for 24 h and a 40% growth inhibition with 5 mM ALA. The strange shape of the viability curves obtained in both cases, with a restoration of control viability with 2 mM ALA, cannot be explained.

Bottom Line: Because apoptosis is usually associated with DNA fragmentation, the DNA of ALA treated and untreated cells were analyzed.To elucidate the mechanisms of ALA induced apoptosis, we examined its effect on p53 expression.CDK2 and CDK4 protein levels were reduced after ALA treatment at physiological concentrations.

View Article: PubMed Central - HTML - PubMed

Affiliation: Centro de Investigaciones sobre Porfirinas y Porfirias, Argentine National Research Council (CONICET), Department of Biological Chemistry, FCEN, University of Buenos Aires, Argentina. desiervi@qb.fcen.uba.ar

ABSTRACT

Background: Acute Intermittent Porphyria is a genetic disorder of heme metabolism, characterized by increased levels of porphyrin precursors, delta-aminolevulinic acid (ALA) and porphobilinogen (PBG). ALA has been reported to generate reactive oxygen species and to cause oxidative damage to proteins, subcellular structures and DNA. It is known that oxidative stress can induce apoptosis. The aim of this work was to study the cytotoxic effect of ALA on two hepatocarcinoma cell lines.

Results: We have determined the impact of ALA on HEP G2 and HEP 3B hepatocarcinoma cell lines survival as measured by the MTT assay. ALA proved to be cytotoxic in both cell lines however; HEP G2 was more sensitive to ALA than HEP 3B. Addition of hemin or glucose diminished ALA cytotoxicity in HEP G2 cells; instead it was enhanced in HEP 3B cells. Because apoptosis is usually associated with DNA fragmentation, the DNA of ALA treated and untreated cells were analyzed. The characteristic pattern of DNA fragmentation ladders was observed in ALA treated cells. To elucidate the mechanisms of ALA induced apoptosis, we examined its effect on p53 expression. No changes in p53 mRNA levels were observed after exposure of both cell lines to ALA for 24 h. CDK2 and CDK4 protein levels were reduced after ALA treatment at physiological concentrations.

Show MeSH
Related in: MedlinePlus