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Camel milk modulates the expression of aryl hydrocarbon receptor-regulated genes, Cyp1a1, Nqo1, and Gsta1, in murine hepatoma Hepa 1c1c7 cells.

Korashy HM, El Gendy MA, Alhaider AA, El-Kadi AO - J. Biomed. Biotechnol. (2012)

Bottom Line: In addition, camel milk significantly decreased the xenobiotic responsive element (XRE)-dependent luciferase activity, suggesting a transcriptional mechanism is involved.Furthermore, this inhibitory effect of camel milk was associated with a proportional increase in heme oxygenase 1.In conclusion, camel milk modulates the expression of Cyp1a1, Nqo1, and Gsta1 at the transcriptional and posttranscriptional levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, 11451 Riyadh, Saudi Arabia.

ABSTRACT
There is a traditional belief in the Middle East that camel milk may aid in prevention and treatment of numerous cases of cancer yet, the exact mechanism was not investigated. Therefore, we examined the ability of camel milk to modulate the expression of a well-known cancer-activating gene, Cytochrome P450 1a1 (Cyp1a1), and cancer-protective genes, NAD(P)H:quinone oxidoreductase 1 (Nqo1) and glutathione S-transferase a1 (Gsta1), in murine hepatoma Hepa 1c1c7 cell line. Our results showed that camel milk significantly inhibited the induction of Cyp1a1 gene expression by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), the most potent Cyp1a1 inducer and known carcinogenic chemical, at mRNA, protein, and activity levels in a concentration-dependent manner. In addition, camel milk significantly decreased the xenobiotic responsive element (XRE)-dependent luciferase activity, suggesting a transcriptional mechanism is involved. Furthermore, this inhibitory effect of camel milk was associated with a proportional increase in heme oxygenase 1. On the other hand, camel milk significantly induced Nqo1 and Gsta1 mRNA expression level in a concentration-dependent fashion. The RNA synthesis inhibitor, actinomycin D, completely blocked the induction of Nqo1 mRNA by camel milk suggesting the requirement of de novo RNA synthesis through a transcriptional mechanism. In conclusion, camel milk modulates the expression of Cyp1a1, Nqo1, and Gsta1 at the transcriptional and posttranscriptional levels.

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Effect of camel milk on XRE-dependent luciferase activity. Hepa 1c1c7 cells were transiently cotransfected with XRE-driven luciferase reporter plasmid pGudLuc 1.1. and renilla luciferase control plasmid pRL-CMV. Cells were treated with sterile water or camel milk (fat free, 100 μL/mL) 30 min before the addition of TCDD (1 nM) for an additional 24 h. Cells were lysed and luciferase activity was reported as relative light unit (RLU) of firefly luciferase to renilla luciferase (Fluc/Rluc) (mean ± S.E.M., n = 4).   +P < 0.05 compared control (sterile water-treated cells),   *P < 0.05 compared with TCDD-treated cells.
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fig5: Effect of camel milk on XRE-dependent luciferase activity. Hepa 1c1c7 cells were transiently cotransfected with XRE-driven luciferase reporter plasmid pGudLuc 1.1. and renilla luciferase control plasmid pRL-CMV. Cells were treated with sterile water or camel milk (fat free, 100 μL/mL) 30 min before the addition of TCDD (1 nM) for an additional 24 h. Cells were lysed and luciferase activity was reported as relative light unit (RLU) of firefly luciferase to renilla luciferase (Fluc/Rluc) (mean ± S.E.M., n = 4).   +P < 0.05 compared control (sterile water-treated cells),   *P < 0.05 compared with TCDD-treated cells.

Mentions: To explore the ability of fat-free camel milk to inhibit the AhR-dependent reporter gene expression, Hepa 1c1c7 cells were transiently cotransfected with the XRE-dependent luciferase reporter gene and renilla luciferase vector, which was used for normalization of transfection efficiency. Cells were then incubated for 24 h with TCDD (I nM) in the presence and absence of camel milk (100 μL/mL). Our results showed that TCDD significantly induced XRE-dependent luciferase activity by approximately 7-fold (Figure 5). Importantly, camel milk (fat-free, 100 μL/mL) completely blocked the induction of XRE-dependent luciferase activity by TCDD to its control level (Figure 5).


Camel milk modulates the expression of aryl hydrocarbon receptor-regulated genes, Cyp1a1, Nqo1, and Gsta1, in murine hepatoma Hepa 1c1c7 cells.

Korashy HM, El Gendy MA, Alhaider AA, El-Kadi AO - J. Biomed. Biotechnol. (2012)

Effect of camel milk on XRE-dependent luciferase activity. Hepa 1c1c7 cells were transiently cotransfected with XRE-driven luciferase reporter plasmid pGudLuc 1.1. and renilla luciferase control plasmid pRL-CMV. Cells were treated with sterile water or camel milk (fat free, 100 μL/mL) 30 min before the addition of TCDD (1 nM) for an additional 24 h. Cells were lysed and luciferase activity was reported as relative light unit (RLU) of firefly luciferase to renilla luciferase (Fluc/Rluc) (mean ± S.E.M., n = 4).   +P < 0.05 compared control (sterile water-treated cells),   *P < 0.05 compared with TCDD-treated cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3345340&req=5

fig5: Effect of camel milk on XRE-dependent luciferase activity. Hepa 1c1c7 cells were transiently cotransfected with XRE-driven luciferase reporter plasmid pGudLuc 1.1. and renilla luciferase control plasmid pRL-CMV. Cells were treated with sterile water or camel milk (fat free, 100 μL/mL) 30 min before the addition of TCDD (1 nM) for an additional 24 h. Cells were lysed and luciferase activity was reported as relative light unit (RLU) of firefly luciferase to renilla luciferase (Fluc/Rluc) (mean ± S.E.M., n = 4).   +P < 0.05 compared control (sterile water-treated cells),   *P < 0.05 compared with TCDD-treated cells.
Mentions: To explore the ability of fat-free camel milk to inhibit the AhR-dependent reporter gene expression, Hepa 1c1c7 cells were transiently cotransfected with the XRE-dependent luciferase reporter gene and renilla luciferase vector, which was used for normalization of transfection efficiency. Cells were then incubated for 24 h with TCDD (I nM) in the presence and absence of camel milk (100 μL/mL). Our results showed that TCDD significantly induced XRE-dependent luciferase activity by approximately 7-fold (Figure 5). Importantly, camel milk (fat-free, 100 μL/mL) completely blocked the induction of XRE-dependent luciferase activity by TCDD to its control level (Figure 5).

Bottom Line: In addition, camel milk significantly decreased the xenobiotic responsive element (XRE)-dependent luciferase activity, suggesting a transcriptional mechanism is involved.Furthermore, this inhibitory effect of camel milk was associated with a proportional increase in heme oxygenase 1.In conclusion, camel milk modulates the expression of Cyp1a1, Nqo1, and Gsta1 at the transcriptional and posttranscriptional levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, 11451 Riyadh, Saudi Arabia.

ABSTRACT
There is a traditional belief in the Middle East that camel milk may aid in prevention and treatment of numerous cases of cancer yet, the exact mechanism was not investigated. Therefore, we examined the ability of camel milk to modulate the expression of a well-known cancer-activating gene, Cytochrome P450 1a1 (Cyp1a1), and cancer-protective genes, NAD(P)H:quinone oxidoreductase 1 (Nqo1) and glutathione S-transferase a1 (Gsta1), in murine hepatoma Hepa 1c1c7 cell line. Our results showed that camel milk significantly inhibited the induction of Cyp1a1 gene expression by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), the most potent Cyp1a1 inducer and known carcinogenic chemical, at mRNA, protein, and activity levels in a concentration-dependent manner. In addition, camel milk significantly decreased the xenobiotic responsive element (XRE)-dependent luciferase activity, suggesting a transcriptional mechanism is involved. Furthermore, this inhibitory effect of camel milk was associated with a proportional increase in heme oxygenase 1. On the other hand, camel milk significantly induced Nqo1 and Gsta1 mRNA expression level in a concentration-dependent fashion. The RNA synthesis inhibitor, actinomycin D, completely blocked the induction of Nqo1 mRNA by camel milk suggesting the requirement of de novo RNA synthesis through a transcriptional mechanism. In conclusion, camel milk modulates the expression of Cyp1a1, Nqo1, and Gsta1 at the transcriptional and posttranscriptional levels.

Show MeSH
Related in: MedlinePlus