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Anti-Cancer Effect of IN-2001 in MDA-MB-231 Human Breast Cancer.

Min KN, Joung KE, Kim DK, Sheen YY - Biomol Ther (Seoul) (2012)

Bottom Line: But their precise mechanism of action has not been elucidated.These events are accompanied by modulating several cell cycle and apoptosis regulatory genes such as CDK inhibitors p21(WAF1) and p27(KIP1) cyclin D1, and other tumor suppressor genes such as cyclin D2.In summary, our data suggest that this histone deacetylase inhibitor, IN-2001, is a novel promising therapeutic agent with potent antitumor effects against human breast cancers.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Ewha Womans University, Seoul 120-750, Republic of Korea.

ABSTRACT
In recent years, inhibition of HDACs has emerged as a potential strategy to reverse aberrant epigenetic changes associated with cancer, and several classes of HDAC inhibitors have been found to have potent and specific anticancer activities in preclinical studies. But their precise mechanism of action has not been elucidated. In this study, a novel synthetic inhibitor of HDAC, 3-(4-dimethylamino phenyl)-N-hydroxy-2-propenamide [IN-2001] was examined for its antitumor activity and the underlying molecular mechanisms of any such activity on human breast cancer cell lines. IN-2001 effectively inhibited cellular HDAC activity (IC50 = 0.585 nM) in MDA-MB-231 human breast cancer cells. IN-2001 caused a significant dose-dependent inhibition of cell proliferation in estrogen receptor (ER) negative MDA-MB-231 human breast cancer cells. Cell cycle analysis revealed that the gowth inhibitory effects of IN-2001 might be attributed to cell cycle arrest at G0/G1 and/or G2/Mphase and subsequent apoptosis in human breast cancer cells. These events are accompanied by modulating several cell cycle and apoptosis regulatory genes such as CDK inhibitors p21(WAF1) and p27(KIP1) cyclin D1, and other tumor suppressor genes such as cyclin D2. Collectively, IN-2001 inhibited cell proliferation and induced apoptosis in human breast cancer cells and these findings may provide new therapeutic approaches, combination of antiestrogen together with a HDAC inhibitor, in the hormonal therapy-resistant ER-negative breast cancers. In summary, our data suggest that this histone deacetylase inhibitor, IN-2001, is a novel promising therapeutic agent with potent antitumor effects against human breast cancers.

No MeSH data available.


Related in: MedlinePlus

Effect of IN-2001 on the expression of cdk inhibitor. MDA-MB-231 cells were treated with vehicle (0.1% DMSO) or 1 μM IN-2001 for 24 hr.Protein extracts were prepared and 50 μg of protein extracts were separated by 12% SDS-PAGE. Blots were probed with the corresponding antibodies. Actin served as the loading control. For quantification, the band intensity of p21WAF1 and p27KIP1 was normalized to that of Actin and data was expressed as fold induction compared to control group.
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Figure 006: Effect of IN-2001 on the expression of cdk inhibitor. MDA-MB-231 cells were treated with vehicle (0.1% DMSO) or 1 μM IN-2001 for 24 hr.Protein extracts were prepared and 50 μg of protein extracts were separated by 12% SDS-PAGE. Blots were probed with the corresponding antibodies. Actin served as the loading control. For quantification, the band intensity of p21WAF1 and p27KIP1 was normalized to that of Actin and data was expressed as fold induction compared to control group.

Mentions: In the previous study, we found that HDAC inhibitors induced cell cycle arrest. In relation to cell cycle arrest, we examined the effects of HDAC inhibitorson the cell cycle regulatory proteins, such as cyclins and cyclin dependent kinase (cdk) inhibitors. MDA-MB-231 cells were treated with vehicle (0.1% DMSO) or 1 μM IN-2001 for 24 hr. And then the expression of cdk inhibitors, such as p21WAF1 and p27KIP1 was examined by RT-PCR and western blot analysis. As shown in Fig. 5 in MDA-MB-231 cells, IN2001, and SAHA slightly increased p21WAF1 mRNA level. In contrast, p21WAF1 protein level was significantly up-regulated by all kinds of IN-2001 (Fig. 6). IN-2001, and SAHA treatment showed 1.9-fold, and 1.4-fold in-crease in p21WAF1 protein level, respectively. In addition, p27KIP1 protein level was also increased to 2.6-fold, and 1.5-fold with IN2001, and SAHA, respectively.


Anti-Cancer Effect of IN-2001 in MDA-MB-231 Human Breast Cancer.

Min KN, Joung KE, Kim DK, Sheen YY - Biomol Ther (Seoul) (2012)

Effect of IN-2001 on the expression of cdk inhibitor. MDA-MB-231 cells were treated with vehicle (0.1% DMSO) or 1 μM IN-2001 for 24 hr.Protein extracts were prepared and 50 μg of protein extracts were separated by 12% SDS-PAGE. Blots were probed with the corresponding antibodies. Actin served as the loading control. For quantification, the band intensity of p21WAF1 and p27KIP1 was normalized to that of Actin and data was expressed as fold induction compared to control group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3794529&req=5

Figure 006: Effect of IN-2001 on the expression of cdk inhibitor. MDA-MB-231 cells were treated with vehicle (0.1% DMSO) or 1 μM IN-2001 for 24 hr.Protein extracts were prepared and 50 μg of protein extracts were separated by 12% SDS-PAGE. Blots were probed with the corresponding antibodies. Actin served as the loading control. For quantification, the band intensity of p21WAF1 and p27KIP1 was normalized to that of Actin and data was expressed as fold induction compared to control group.
Mentions: In the previous study, we found that HDAC inhibitors induced cell cycle arrest. In relation to cell cycle arrest, we examined the effects of HDAC inhibitorson the cell cycle regulatory proteins, such as cyclins and cyclin dependent kinase (cdk) inhibitors. MDA-MB-231 cells were treated with vehicle (0.1% DMSO) or 1 μM IN-2001 for 24 hr. And then the expression of cdk inhibitors, such as p21WAF1 and p27KIP1 was examined by RT-PCR and western blot analysis. As shown in Fig. 5 in MDA-MB-231 cells, IN2001, and SAHA slightly increased p21WAF1 mRNA level. In contrast, p21WAF1 protein level was significantly up-regulated by all kinds of IN-2001 (Fig. 6). IN-2001, and SAHA treatment showed 1.9-fold, and 1.4-fold in-crease in p21WAF1 protein level, respectively. In addition, p27KIP1 protein level was also increased to 2.6-fold, and 1.5-fold with IN2001, and SAHA, respectively.

Bottom Line: But their precise mechanism of action has not been elucidated.These events are accompanied by modulating several cell cycle and apoptosis regulatory genes such as CDK inhibitors p21(WAF1) and p27(KIP1) cyclin D1, and other tumor suppressor genes such as cyclin D2.In summary, our data suggest that this histone deacetylase inhibitor, IN-2001, is a novel promising therapeutic agent with potent antitumor effects against human breast cancers.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Ewha Womans University, Seoul 120-750, Republic of Korea.

ABSTRACT
In recent years, inhibition of HDACs has emerged as a potential strategy to reverse aberrant epigenetic changes associated with cancer, and several classes of HDAC inhibitors have been found to have potent and specific anticancer activities in preclinical studies. But their precise mechanism of action has not been elucidated. In this study, a novel synthetic inhibitor of HDAC, 3-(4-dimethylamino phenyl)-N-hydroxy-2-propenamide [IN-2001] was examined for its antitumor activity and the underlying molecular mechanisms of any such activity on human breast cancer cell lines. IN-2001 effectively inhibited cellular HDAC activity (IC50 = 0.585 nM) in MDA-MB-231 human breast cancer cells. IN-2001 caused a significant dose-dependent inhibition of cell proliferation in estrogen receptor (ER) negative MDA-MB-231 human breast cancer cells. Cell cycle analysis revealed that the gowth inhibitory effects of IN-2001 might be attributed to cell cycle arrest at G0/G1 and/or G2/Mphase and subsequent apoptosis in human breast cancer cells. These events are accompanied by modulating several cell cycle and apoptosis regulatory genes such as CDK inhibitors p21(WAF1) and p27(KIP1) cyclin D1, and other tumor suppressor genes such as cyclin D2. Collectively, IN-2001 inhibited cell proliferation and induced apoptosis in human breast cancer cells and these findings may provide new therapeutic approaches, combination of antiestrogen together with a HDAC inhibitor, in the hormonal therapy-resistant ER-negative breast cancers. In summary, our data suggest that this histone deacetylase inhibitor, IN-2001, is a novel promising therapeutic agent with potent antitumor effects against human breast cancers.

No MeSH data available.


Related in: MedlinePlus