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CArG-driven GADD45α activated by resveratrol inhibits lung cancer cells.

Shi Q, Geldenhuys W, Sutariya V, Bishayee A, Patel I, Bhatia D - Genes Cancer (2015)

Bottom Line: Therefore, induction of exogenous GADD45α resulted in growth inhibition.Since natural promoter may have antagonized effects, we tested synthetic promoter that contains either five, six or nine repeats of CArG elements essential in the Egr-1 promoter to drive the expression of GADD45α upon resveratrol treatment.Further analysis confirmed that both synthetic promoter and natural Egr-1 promoter were able to "turn on" the expression of GADD45α when combined with resveratrol, and subsequently led to suppression of cell proliferation and apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, Northeast Ohio Medical University, Rootstown, Ohio, USA ; School of Biomedical Sciences, Kent State University, Kent, Ohio, USA.

ABSTRACT
We report anticarcinogenic effects of suicide gene therapy that relies on the use of resveratrol-responsive CArG elements from the Egr-1 promoter to induce GADD45α. In A549 lung cancer cells, endogenous GADD45α was not induced upon resveratrol treatment. Therefore, induction of exogenous GADD45α resulted in growth inhibition. Resveratrol transiently induced Egr-1 through ERK/JNK-ElK-1. Hence, we cloned natural or synthetic Egr-1 promoter upstream of GADD45α cDNA to create a suicide gene therapy vector. Since natural promoter may have antagonized effects, we tested synthetic promoter that contains either five, six or nine repeats of CArG elements essential in the Egr-1 promoter to drive the expression of GADD45α upon resveratrol treatment. Further analysis confirmed that both synthetic promoter and natural Egr-1 promoter were able to "turn on" the expression of GADD45α when combined with resveratrol, and subsequently led to suppression of cell proliferation and apoptosis.

No MeSH data available.


Related in: MedlinePlus

Resveratrol induces Egr-1 expressionA. A549 cells were treated with 100 μM of resveratrol for indicated hours and Egr-1 mRNA levels were measured by real-time RT-PCR. The Egr-1 mRNA expression was adjusted to GAPDH (housekeeping gene) and normalized to untreated control. B. and C. Egr-1 and β-actin (loading control) protein expression was determined by western blotting after indicated resveratrol treatment. D. Fluorescent In Situ Hybridization assay was used to detect Egr-1 mRNA level in non-treated and resveratrol-treated cells. Images were taken on Axio Imager M2 at 400X magnification.
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Figure 1: Resveratrol induces Egr-1 expressionA. A549 cells were treated with 100 μM of resveratrol for indicated hours and Egr-1 mRNA levels were measured by real-time RT-PCR. The Egr-1 mRNA expression was adjusted to GAPDH (housekeeping gene) and normalized to untreated control. B. and C. Egr-1 and β-actin (loading control) protein expression was determined by western blotting after indicated resveratrol treatment. D. Fluorescent In Situ Hybridization assay was used to detect Egr-1 mRNA level in non-treated and resveratrol-treated cells. Images were taken on Axio Imager M2 at 400X magnification.

Mentions: Resveratrol is able to activate Egr-1 expression in several cancers including pancreatic [25] and colorectal cancer [27]. To determine whether resveratrol is an effective inducer of Egr-1 expression in lung cancer, A549 cells were incubated with 100 μM of resveratrol, and mRNA level and protein expression were measured at different time points by real-time RT-PCR and western blotting respectively. The induction of Egr-1 mRNA and protein reached maximum 2 h and 4 h respectively after resveratrol treatment (Figure 1A and 1B). In addition, the expression of Egr-1 protein was measured after cells were treated with 0, 25, 50 and 100 μM of resveratrol for 6 h, and a dose-dependent induction was detected (Figure 1C). Fluorescence in situ hybridization (FISH) assay reinforced that Egr-1 mRNA level was significantly induced 2 h after resveratrol exposure (Figure 1D). These results suggest that resveratrol is able to induce Egr-1 expression rapidly and transiently.


CArG-driven GADD45α activated by resveratrol inhibits lung cancer cells.

Shi Q, Geldenhuys W, Sutariya V, Bishayee A, Patel I, Bhatia D - Genes Cancer (2015)

Resveratrol induces Egr-1 expressionA. A549 cells were treated with 100 μM of resveratrol for indicated hours and Egr-1 mRNA levels were measured by real-time RT-PCR. The Egr-1 mRNA expression was adjusted to GAPDH (housekeeping gene) and normalized to untreated control. B. and C. Egr-1 and β-actin (loading control) protein expression was determined by western blotting after indicated resveratrol treatment. D. Fluorescent In Situ Hybridization assay was used to detect Egr-1 mRNA level in non-treated and resveratrol-treated cells. Images were taken on Axio Imager M2 at 400X magnification.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
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getmorefigures.php?uid=PMC4482243&req=5

Figure 1: Resveratrol induces Egr-1 expressionA. A549 cells were treated with 100 μM of resveratrol for indicated hours and Egr-1 mRNA levels were measured by real-time RT-PCR. The Egr-1 mRNA expression was adjusted to GAPDH (housekeeping gene) and normalized to untreated control. B. and C. Egr-1 and β-actin (loading control) protein expression was determined by western blotting after indicated resveratrol treatment. D. Fluorescent In Situ Hybridization assay was used to detect Egr-1 mRNA level in non-treated and resveratrol-treated cells. Images were taken on Axio Imager M2 at 400X magnification.
Mentions: Resveratrol is able to activate Egr-1 expression in several cancers including pancreatic [25] and colorectal cancer [27]. To determine whether resveratrol is an effective inducer of Egr-1 expression in lung cancer, A549 cells were incubated with 100 μM of resveratrol, and mRNA level and protein expression were measured at different time points by real-time RT-PCR and western blotting respectively. The induction of Egr-1 mRNA and protein reached maximum 2 h and 4 h respectively after resveratrol treatment (Figure 1A and 1B). In addition, the expression of Egr-1 protein was measured after cells were treated with 0, 25, 50 and 100 μM of resveratrol for 6 h, and a dose-dependent induction was detected (Figure 1C). Fluorescence in situ hybridization (FISH) assay reinforced that Egr-1 mRNA level was significantly induced 2 h after resveratrol exposure (Figure 1D). These results suggest that resveratrol is able to induce Egr-1 expression rapidly and transiently.

Bottom Line: Therefore, induction of exogenous GADD45α resulted in growth inhibition.Since natural promoter may have antagonized effects, we tested synthetic promoter that contains either five, six or nine repeats of CArG elements essential in the Egr-1 promoter to drive the expression of GADD45α upon resveratrol treatment.Further analysis confirmed that both synthetic promoter and natural Egr-1 promoter were able to "turn on" the expression of GADD45α when combined with resveratrol, and subsequently led to suppression of cell proliferation and apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, Northeast Ohio Medical University, Rootstown, Ohio, USA ; School of Biomedical Sciences, Kent State University, Kent, Ohio, USA.

ABSTRACT
We report anticarcinogenic effects of suicide gene therapy that relies on the use of resveratrol-responsive CArG elements from the Egr-1 promoter to induce GADD45α. In A549 lung cancer cells, endogenous GADD45α was not induced upon resveratrol treatment. Therefore, induction of exogenous GADD45α resulted in growth inhibition. Resveratrol transiently induced Egr-1 through ERK/JNK-ElK-1. Hence, we cloned natural or synthetic Egr-1 promoter upstream of GADD45α cDNA to create a suicide gene therapy vector. Since natural promoter may have antagonized effects, we tested synthetic promoter that contains either five, six or nine repeats of CArG elements essential in the Egr-1 promoter to drive the expression of GADD45α upon resveratrol treatment. Further analysis confirmed that both synthetic promoter and natural Egr-1 promoter were able to "turn on" the expression of GADD45α when combined with resveratrol, and subsequently led to suppression of cell proliferation and apoptosis.

No MeSH data available.


Related in: MedlinePlus