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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.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.

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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Ilimaquinone and ethylsmenoquinone induce apoptosis in HCT116 cells. (A) HCT116 cells were incubated with the vehicle (DMSO) or either IQ or ESQ for 24 h. Following treatment, cells were harvested and stained with Annexin V-FITC and PI and analyzed using a flow cytometry. The x-axis indicates the Annexin V-FITC intensity, while the y-axis indicates the PI fluorescence; (B) Cell extracts from HCT116 cells treated with either vehicle (DMSO) or the indicated concentrations of IQ and ESQ for 24 h were analyzed by western blotting with anti-caspase 3 and anti-PARP antibodies. 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-f005: Ilimaquinone and ethylsmenoquinone induce apoptosis in HCT116 cells. (A) HCT116 cells were incubated with the vehicle (DMSO) or either IQ or ESQ for 24 h. Following treatment, cells were harvested and stained with Annexin V-FITC and PI and analyzed using a flow cytometry. The x-axis indicates the Annexin V-FITC intensity, while the y-axis indicates the PI fluorescence; (B) Cell extracts from HCT116 cells treated with either vehicle (DMSO) or the indicated concentrations of IQ and ESQ for 24 h were analyzed by western blotting with anti-caspase 3 and anti-PARP antibodies. The blots were re-probed with anti-actin antibodies as a loading control. Results are representative of three independent experiments.

Mentions: Activation of the p53 pathway has been reported to induce apoptosis in various cancer cells [1,2]. Thus, we examined the abilities of ilimaquinone and ethylsmenoquinone to stimulate apoptosis in colon cancer cells. For this purpose, HCT116 cells were incubated with the sesquiterpene quinones, and the number of apoptotic cells was quantified using Annexin V/PI staining. As shown in Figure 5A, cytometric analysis revealed that the percentage of apoptotic cells significantly increased in a concentration-dependent manner. Compared to control cells, the percentage of Annexin V/PI double-positive cells increased from 8.95% to 27.4% and from 8.87% to 32.74% in HCT116 cells treated with 10 μM ilimaquinone and ethylsmenoquinone, respectively (Figure 5A). In addition, western blot analysis showed that treating HCT116 cells with either compound enhanced the proteolytic cleavage of pro-caspase-3, thereby activating caspase-3, which, in turn, catalyzed the cleavage of poly ADP ribose polymerase (PARP), a biochemical marker of apoptosis, (Figure 5B). This suggests that apoptosis contributed to ilimaquinone- and ethylsmenoquinone-induced cell death.


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)

Ilimaquinone and ethylsmenoquinone induce apoptosis in HCT116 cells. (A) HCT116 cells were incubated with the vehicle (DMSO) or either IQ or ESQ for 24 h. Following treatment, cells were harvested and stained with Annexin V-FITC and PI and analyzed using a flow cytometry. The x-axis indicates the Annexin V-FITC intensity, while the y-axis indicates the PI fluorescence; (B) Cell extracts from HCT116 cells treated with either vehicle (DMSO) or the indicated concentrations of IQ and ESQ for 24 h were analyzed by western blotting with anti-caspase 3 and anti-PARP antibodies. 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

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

marinedrugs-13-00543-f005: Ilimaquinone and ethylsmenoquinone induce apoptosis in HCT116 cells. (A) HCT116 cells were incubated with the vehicle (DMSO) or either IQ or ESQ for 24 h. Following treatment, cells were harvested and stained with Annexin V-FITC and PI and analyzed using a flow cytometry. The x-axis indicates the Annexin V-FITC intensity, while the y-axis indicates the PI fluorescence; (B) Cell extracts from HCT116 cells treated with either vehicle (DMSO) or the indicated concentrations of IQ and ESQ for 24 h were analyzed by western blotting with anti-caspase 3 and anti-PARP antibodies. The blots were re-probed with anti-actin antibodies as a loading control. Results are representative of three independent experiments.
Mentions: Activation of the p53 pathway has been reported to induce apoptosis in various cancer cells [1,2]. Thus, we examined the abilities of ilimaquinone and ethylsmenoquinone to stimulate apoptosis in colon cancer cells. For this purpose, HCT116 cells were incubated with the sesquiterpene quinones, and the number of apoptotic cells was quantified using Annexin V/PI staining. As shown in Figure 5A, cytometric analysis revealed that the percentage of apoptotic cells significantly increased in a concentration-dependent manner. Compared to control cells, the percentage of Annexin V/PI double-positive cells increased from 8.95% to 27.4% and from 8.87% to 32.74% in HCT116 cells treated with 10 μM ilimaquinone and ethylsmenoquinone, respectively (Figure 5A). In addition, western blot analysis showed that treating HCT116 cells with either compound enhanced the proteolytic cleavage of pro-caspase-3, thereby activating caspase-3, which, in turn, catalyzed the cleavage of poly ADP ribose polymerase (PARP), a biochemical marker of apoptosis, (Figure 5B). This suggests that apoptosis contributed to ilimaquinone- and ethylsmenoquinone-induced cell death.

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.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.

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