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Activation of p53 with ilimaquinone and ethylsmenoquinone, marine sponge metabolites, induces apoptosis and autophagy in colon cancer cells.

Lee HY, Chung KJ, Hwang IH, Gwak J, Park S, Ju BG, Yun E, Kim DE, Chung YH, Na M, Song GY, Oh S - Mar Drugs (2015)

Bottom Line: Here, we used a cell-based reporter system for the detection of p53 response transcription to identify the marine sponge metabolites, ilimaquinone and ethylsmenoquinone, as activators of the p53 pathway.In addition, ilimaquinone and ethylsmenoquinone induced G2/M cell cycle arrest and increased caspase-3 cleavage and the population of cells that positively stained with Annexin V-FITC, both of which are typical biochemical markers of apoptosis.Furthermore, autophagy was elicited by both compounds, as indicated by microtubule-associated protein 1 light chain 3 (LC3) puncta formations and LC3-II turnover in HCT116 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Bio and Fermentation Convergence Technology, Kookmin University, Seoul 136-702, Korea. IHyunYoungg@gmail.com.

ABSTRACT
The tumor suppressor, p53, plays an essential role in the cellular response to stress through regulating the expression of genes involved in cell cycle arrest, apoptosis and autophagy. Here, we used a cell-based reporter system for the detection of p53 response transcription to identify the marine sponge metabolites, ilimaquinone and ethylsmenoquinone, as activators of the p53 pathway. We demonstrated that ilimaquinone and ethylsmenoquinone efficiently stabilize the p53 protein through promotion of p53 phosphorylation at Ser15 in both HCT116 and RKO colon cancer cells. Moreover, both compounds upregulate the expression of p21WAF1/CIP1, a p53-dependent gene, and suppress proliferation of colon cancer cells. In addition, ilimaquinone and ethylsmenoquinone induced G2/M cell cycle arrest and increased caspase-3 cleavage and the population of cells that positively stained with Annexin V-FITC, both of which are typical biochemical markers of apoptosis. Furthermore, autophagy was elicited by both compounds, as indicated by microtubule-associated protein 1 light chain 3 (LC3) puncta formations and LC3-II turnover in HCT116 cells. Our findings suggest that ilimaquinone and ethylsmenoquinone exert their anti-cancer activity by activation of the p53 pathway and may have significant potential as chemo-preventive and therapeutic agents for human colon cancer.

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The effects of ilimaquinone and ethylsmenoquinone on the level of p21WAF1/CIP1 in colon cancer cells. (A,B) Semi-quantitative RT-PCR for p21WAF1/CIP1 and GAPDH were performed using total RNA prepared from HCT116 and RKO cells that were incubated with vehicle (DMSO) or the indicated concentrations of IQ and ESQ; the results are representative of three independent experiments; (C,D) Cell extracts from HCT116 and RKO cells treated with either vehicle (DMSO) or IQ and ESQ were analyzed by western blotting with anti-p21WAF1/CIP1 antibodies; In C and D, the blots were re-probed with anti-actin antibodies as a loading control. Results are representative of three independent experiments.
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marinedrugs-13-00543-f003: The effects of ilimaquinone and ethylsmenoquinone on the level of p21WAF1/CIP1 in colon cancer cells. (A,B) Semi-quantitative RT-PCR for p21WAF1/CIP1 and GAPDH were performed using total RNA prepared from HCT116 and RKO cells that were incubated with vehicle (DMSO) or the indicated concentrations of IQ and ESQ; the results are representative of three independent experiments; (C,D) Cell extracts from HCT116 and RKO cells treated with either vehicle (DMSO) or IQ and ESQ were analyzed by western blotting with anti-p21WAF1/CIP1 antibodies; In C and D, the blots were re-probed with anti-actin antibodies as a loading control. Results are representative of three independent experiments.

Mentions: We next tested the effects of ilimaquinone and ethylsmenoquinone on the p53-dependent gene expression in colon cancer cells. A semi-quantitative RT-PCR assay demonstrated that the mRNA levels of p21WAF1/CIP1, a known target gene of p53, were elevated by treatment with ilimaquinone and ethylsmenoquinone in both HCT116 and RKO cells (Figure 3A,B). In conjunction with the resulting data, we observed that treatment with ilimaquinone and ethylsmenoquinone led to an increase in the level of p21WAF1/CIP1 protein (Figure 3C,D). Taken together, these results suggest that ilimaquinone and ethylsmenoquinone are specific activators of the p53 pathway.


Activation of p53 with ilimaquinone and ethylsmenoquinone, marine sponge metabolites, induces apoptosis and autophagy in colon cancer cells.

Lee HY, Chung KJ, Hwang IH, Gwak J, Park S, Ju BG, Yun E, Kim DE, Chung YH, Na M, Song GY, Oh S - Mar Drugs (2015)

The effects of ilimaquinone and ethylsmenoquinone on the level of p21WAF1/CIP1 in colon cancer cells. (A,B) Semi-quantitative RT-PCR for p21WAF1/CIP1 and GAPDH were performed using total RNA prepared from HCT116 and RKO cells that were incubated with vehicle (DMSO) or the indicated concentrations of IQ and ESQ; the results are representative of three independent experiments; (C,D) Cell extracts from HCT116 and RKO cells treated with either vehicle (DMSO) or IQ and ESQ were analyzed by western blotting with anti-p21WAF1/CIP1 antibodies; In C and D, the blots were re-probed with anti-actin antibodies as a loading control. Results are representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4306951&req=5

marinedrugs-13-00543-f003: The effects of ilimaquinone and ethylsmenoquinone on the level of p21WAF1/CIP1 in colon cancer cells. (A,B) Semi-quantitative RT-PCR for p21WAF1/CIP1 and GAPDH were performed using total RNA prepared from HCT116 and RKO cells that were incubated with vehicle (DMSO) or the indicated concentrations of IQ and ESQ; the results are representative of three independent experiments; (C,D) Cell extracts from HCT116 and RKO cells treated with either vehicle (DMSO) or IQ and ESQ were analyzed by western blotting with anti-p21WAF1/CIP1 antibodies; In C and D, the blots were re-probed with anti-actin antibodies as a loading control. Results are representative of three independent experiments.
Mentions: We next tested the effects of ilimaquinone and ethylsmenoquinone on the p53-dependent gene expression in colon cancer cells. A semi-quantitative RT-PCR assay demonstrated that the mRNA levels of p21WAF1/CIP1, a known target gene of p53, were elevated by treatment with ilimaquinone and ethylsmenoquinone in both HCT116 and RKO cells (Figure 3A,B). In conjunction with the resulting data, we observed that treatment with ilimaquinone and ethylsmenoquinone led to an increase in the level of p21WAF1/CIP1 protein (Figure 3C,D). Taken together, these results suggest that ilimaquinone and ethylsmenoquinone are specific activators of the p53 pathway.

Bottom Line: Here, we used a cell-based reporter system for the detection of p53 response transcription to identify the marine sponge metabolites, ilimaquinone and ethylsmenoquinone, as activators of the p53 pathway.In addition, ilimaquinone and ethylsmenoquinone induced G2/M cell cycle arrest and increased caspase-3 cleavage and the population of cells that positively stained with Annexin V-FITC, both of which are typical biochemical markers of apoptosis.Furthermore, autophagy was elicited by both compounds, as indicated by microtubule-associated protein 1 light chain 3 (LC3) puncta formations and LC3-II turnover in HCT116 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Bio and Fermentation Convergence Technology, Kookmin University, Seoul 136-702, Korea. IHyunYoungg@gmail.com.

ABSTRACT
The tumor suppressor, p53, plays an essential role in the cellular response to stress through regulating the expression of genes involved in cell cycle arrest, apoptosis and autophagy. Here, we used a cell-based reporter system for the detection of p53 response transcription to identify the marine sponge metabolites, ilimaquinone and ethylsmenoquinone, as activators of the p53 pathway. We demonstrated that ilimaquinone and ethylsmenoquinone efficiently stabilize the p53 protein through promotion of p53 phosphorylation at Ser15 in both HCT116 and RKO colon cancer cells. Moreover, both compounds upregulate the expression of p21WAF1/CIP1, a p53-dependent gene, and suppress proliferation of colon cancer cells. In addition, ilimaquinone and ethylsmenoquinone induced G2/M cell cycle arrest and increased caspase-3 cleavage and the population of cells that positively stained with Annexin V-FITC, both of which are typical biochemical markers of apoptosis. Furthermore, autophagy was elicited by both compounds, as indicated by microtubule-associated protein 1 light chain 3 (LC3) puncta formations and LC3-II turnover in HCT116 cells. Our findings suggest that ilimaquinone and ethylsmenoquinone exert their anti-cancer activity by activation of the p53 pathway and may have significant potential as chemo-preventive and therapeutic agents for human colon cancer.

Show MeSH
Related in: MedlinePlus