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Antineoplastic effects of gamma linolenic Acid on hepatocellular carcinoma cell lines.

Itoh S, Taketomi A, Harimoto N, Tsujita E, Rikimaru T, Shirabe K, Shimada M, Maehara Y - J Clin Biochem Nutr (2010)

Bottom Line: Cell proliferation and reactive oxygen species (ROS) generation including lipid peroxidation and apoptosis were compared.GLA treatment significantly reduced cell proliferation, generated ROS, and induced apoptosis.Furthermore, chromium mesoporphyrin (CrMP), an inhibitor of HO activity, significantly potentiated GLA cytotoxicity.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery and Science, Graduate School of Medical Sciences, Kyushu University, Higashi-ku, Fukuoka 812-8582, Japan.

ABSTRACT
The aim of this study was to investigate the effect and the mechanism of gamma linolenic acid (GLA) treatment on human hepatocellular (HCC) cell lines. The human HCC cell line HuH7 was exposed to GLA. Cell proliferation and reactive oxygen species (ROS) generation including lipid peroxidation and apoptosis were compared. We then used a cDNA microarray analysis to investigate the molecular changes induced by GLA. GLA treatment significantly reduced cell proliferation, generated ROS, and induced apoptosis. After 24 h exposure of Huh7 cells to GLA, we identified several genes encoding the antioxidant proteins to be upregulated: heme oxygenase-1 (HO-1), aldo-keto reductase 1 family C1 (AKR1C1), C4 (AKR1C4), and thioredoxin (Trx). The HO-1 protein levels were overexpressed in Huh7 cells after GLA exposure using a Western blot analysis. Furthermore, chromium mesoporphyrin (CrMP), an inhibitor of HO activity, significantly potentiated GLA cytotoxicity. GLA treatment has induced cell growth inhibition, ROS generation including lipid peroxidation, and HO-1 production for antioxidant protection against oxidative stress caused by GLA in Huh7 cells. GLA treatment should be considered as a therapeutic modality in patients with advanced HCC.

No MeSH data available.


Related in: MedlinePlus

Gene induction in the presence of GLA for 24 h using quantitative real-time RT-PCR. (a) up-regulated genes (b) down-regulated genes. White bars, cDNA microarray fold change; Black bars, real time RT-PCR fold change. (c) GLA induced HO-1 protein in Huh7 cells. Huh7 cells were untreated (control) or with 250 µM GLA (GLA) for 12, 24 and 48 h. HO-1 and β-actin proteins were detected by western blotting.
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Figure 5: Gene induction in the presence of GLA for 24 h using quantitative real-time RT-PCR. (a) up-regulated genes (b) down-regulated genes. White bars, cDNA microarray fold change; Black bars, real time RT-PCR fold change. (c) GLA induced HO-1 protein in Huh7 cells. Huh7 cells were untreated (control) or with 250 µM GLA (GLA) for 12, 24 and 48 h. HO-1 and β-actin proteins were detected by western blotting.

Mentions: The expression of alteration of 12,814 genes was analyzed by cDNA microarray between GLA treated and untreated Huh7 cells for 24 h. Table 2a and 2b demonstrate genes which were up-regulated >2-fold and down-regulated ≤2-fold. Stress response genes and antioxidative genes were mainly up-regulated in the presence of treatment with GLA for 24 h. The most induced gene was heme oxygenase-1 (HO-1). Cell cycle related genes encoding proliferating cell nuclear antigen (PCNA) and S-phase kinase-associated protein 2 (Skp2) were down regulated. To comfirmed these results, changes in mRNA expression of different genes were analyzed by quantitative real-time RT-PCR. We tested the expression of PCNA, Skp2, trefoil factor 3 (TFF3), matrix metalloproteinase 11 (MMP11), aldo-keto reductase family 1, member C1 (AKR1C1), aldo-keto reductase family 1, member C4 (AKR1C4), Thioredoxin (Trx) and Thioredoxin reductase1 1 (TrxR1). All the studied genes were induced over 2-fold (Fig. 5a and 5b). The most induced gene was HO-1 in the presence of treatment with GLA for 24 h, so we investigated the protein expression of HO-1 using western blot analysis. As Fig. 5c shown, HO-1 protein was overexpressed for 12, 24 and 48 h in Huh7 cells.


Antineoplastic effects of gamma linolenic Acid on hepatocellular carcinoma cell lines.

Itoh S, Taketomi A, Harimoto N, Tsujita E, Rikimaru T, Shirabe K, Shimada M, Maehara Y - J Clin Biochem Nutr (2010)

Gene induction in the presence of GLA for 24 h using quantitative real-time RT-PCR. (a) up-regulated genes (b) down-regulated genes. White bars, cDNA microarray fold change; Black bars, real time RT-PCR fold change. (c) GLA induced HO-1 protein in Huh7 cells. Huh7 cells were untreated (control) or with 250 µM GLA (GLA) for 12, 24 and 48 h. HO-1 and β-actin proteins were detected by western blotting.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 5: Gene induction in the presence of GLA for 24 h using quantitative real-time RT-PCR. (a) up-regulated genes (b) down-regulated genes. White bars, cDNA microarray fold change; Black bars, real time RT-PCR fold change. (c) GLA induced HO-1 protein in Huh7 cells. Huh7 cells were untreated (control) or with 250 µM GLA (GLA) for 12, 24 and 48 h. HO-1 and β-actin proteins were detected by western blotting.
Mentions: The expression of alteration of 12,814 genes was analyzed by cDNA microarray between GLA treated and untreated Huh7 cells for 24 h. Table 2a and 2b demonstrate genes which were up-regulated >2-fold and down-regulated ≤2-fold. Stress response genes and antioxidative genes were mainly up-regulated in the presence of treatment with GLA for 24 h. The most induced gene was heme oxygenase-1 (HO-1). Cell cycle related genes encoding proliferating cell nuclear antigen (PCNA) and S-phase kinase-associated protein 2 (Skp2) were down regulated. To comfirmed these results, changes in mRNA expression of different genes were analyzed by quantitative real-time RT-PCR. We tested the expression of PCNA, Skp2, trefoil factor 3 (TFF3), matrix metalloproteinase 11 (MMP11), aldo-keto reductase family 1, member C1 (AKR1C1), aldo-keto reductase family 1, member C4 (AKR1C4), Thioredoxin (Trx) and Thioredoxin reductase1 1 (TrxR1). All the studied genes were induced over 2-fold (Fig. 5a and 5b). The most induced gene was HO-1 in the presence of treatment with GLA for 24 h, so we investigated the protein expression of HO-1 using western blot analysis. As Fig. 5c shown, HO-1 protein was overexpressed for 12, 24 and 48 h in Huh7 cells.

Bottom Line: Cell proliferation and reactive oxygen species (ROS) generation including lipid peroxidation and apoptosis were compared.GLA treatment significantly reduced cell proliferation, generated ROS, and induced apoptosis.Furthermore, chromium mesoporphyrin (CrMP), an inhibitor of HO activity, significantly potentiated GLA cytotoxicity.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery and Science, Graduate School of Medical Sciences, Kyushu University, Higashi-ku, Fukuoka 812-8582, Japan.

ABSTRACT
The aim of this study was to investigate the effect and the mechanism of gamma linolenic acid (GLA) treatment on human hepatocellular (HCC) cell lines. The human HCC cell line HuH7 was exposed to GLA. Cell proliferation and reactive oxygen species (ROS) generation including lipid peroxidation and apoptosis were compared. We then used a cDNA microarray analysis to investigate the molecular changes induced by GLA. GLA treatment significantly reduced cell proliferation, generated ROS, and induced apoptosis. After 24 h exposure of Huh7 cells to GLA, we identified several genes encoding the antioxidant proteins to be upregulated: heme oxygenase-1 (HO-1), aldo-keto reductase 1 family C1 (AKR1C1), C4 (AKR1C4), and thioredoxin (Trx). The HO-1 protein levels were overexpressed in Huh7 cells after GLA exposure using a Western blot analysis. Furthermore, chromium mesoporphyrin (CrMP), an inhibitor of HO activity, significantly potentiated GLA cytotoxicity. GLA treatment has induced cell growth inhibition, ROS generation including lipid peroxidation, and HO-1 production for antioxidant protection against oxidative stress caused by GLA in Huh7 cells. GLA treatment should be considered as a therapeutic modality in patients with advanced HCC.

No MeSH data available.


Related in: MedlinePlus