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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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Glucose effect on ALA toxicity. The effect of Glucose was determined by exposing HEP G2 and HEP 3B cells to 0.5 mM ALA without (white) or with 2 mg/ml glucose (grey) for 24 h before the MTT assay was performed. Controls were grown with 2 mg/ml glucose alone (black). 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 3: Glucose effect on ALA toxicity. The effect of Glucose was determined by exposing HEP G2 and HEP 3B cells to 0.5 mM ALA without (white) or with 2 mg/ml glucose (grey) for 24 h before the MTT assay was performed. Controls were grown with 2 mg/ml glucose alone (black). Viability was expressed in terms of population growth, as percent of an untreated control population with standard error of the mean (SEM).

Mentions: Alike hemin, simultaneous treatment of the HEP G2 cells with ALA (0.5 mM) and D-glucose (2–3 mg/ml) for 24 h resulted in about two-fold decrease in sensitivity to ALA (Figure 3). And again the same treatment of HEP 3B (ALA and D-glucose) enhanced ALA cytotoxicity.


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)

Glucose effect on ALA toxicity. The effect of Glucose was determined by exposing HEP G2 and HEP 3B cells to 0.5 mM ALA without (white) or with 2 mg/ml glucose (grey) for 24 h before the MTT assay was performed. Controls were grown with 2 mg/ml glucose alone (black). 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

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getmorefigures.php?uid=PMC101407&req=5

Figure 3: Glucose effect on ALA toxicity. The effect of Glucose was determined by exposing HEP G2 and HEP 3B cells to 0.5 mM ALA without (white) or with 2 mg/ml glucose (grey) for 24 h before the MTT assay was performed. Controls were grown with 2 mg/ml glucose alone (black). Viability was expressed in terms of population growth, as percent of an untreated control population with standard error of the mean (SEM).
Mentions: Alike hemin, simultaneous treatment of the HEP G2 cells with ALA (0.5 mM) and D-glucose (2–3 mg/ml) for 24 h resulted in about two-fold decrease in sensitivity to ALA (Figure 3). And again the same treatment of HEP 3B (ALA and D-glucose) enhanced ALA cytotoxicity.

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