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The synthesized 2-(2-fluorophenyl)-6,7-methylenedioxyquinolin-4-one (CHM-1) promoted G2/M arrest through inhibition of CDK1 and induced apoptosis through the mitochondrial-dependent pathway in CT-26 murine colorectal adenocarcinoma cells

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Abstract

Background

In this study, we investigated the effects of 2-(2-fluorophenyl)-6,7-methylenedioxyquinolin-4-one (CHM-1) on cell viability, cell cycle arrest and apoptosis in CT-26 murine colorectal adenocarcinoma cells.

Methods

For determining cell viability, the MTT assay was used. CHM-1 promoted G2/M arrest by PI staining and flow cytometric analysis. Apoptotic cells were evaluated by DAPI staining. We used CDK1 kinase assay, Western blot analysis and caspase activity assays for examining the CDK1 activity and proteins correlated with apoptosis and cell cycle arrest. The in vivo anti-tumor effects of CHM-1-P were evaluated in BALB/c mice inoculated with CT-26 cells orthotopic model.

Results

CHM-1 induced CT-26 cell viability inhibition and morphologic changes in a dose-dependent and time-dependent manner and the approximate IC50 was 742.36 nM. CHM-1 induced significant G2/M arrest and apoptosis in CT-26 cells. CHM-1 inhibited the CDK1 activity and decreased CDK1, Cyclin A, Cyclin B protein levels. CHM-1 induced apoptosis in CT-26 cells and promoted increasing of cytosolic cytochrome c, AIF, Bax, BAD, cleavage of pro-caspase-9, and -3. The significant reduction of caspase-9 and -3 activity and increasing the viable CT-26 cells after pretreated with caspase-9 and -3 inhibitor indicated that CHM-1-induced apoptosis was mainly mediated a mitochondria-dependent pathway. CHM-1-P improved mice survival rate, and enlargement of the spleen and liver metastasis were significantly reduced in groups treated with either 10 mg/kg and 30 mg/kg of CHM-1-P and 5-FU in comparison to these of CT-26/BALB/c mice.

Conclusions

Taken together, CHM-1 acted against colorectal adenocarcinoma cells in vitro via G2/M arrest and apoptosis, and CHM-1-P inhibited tumor growth in vivo.

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Acknowledgments

This study was supported by a grant from the CMU97-238 of China Medical University, Taiwan and grants from the NSC 97-2323-B-039-001 and NSC 97-2320-B-039-004-MY3 of National Science Council, Taiwan.

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Authors and Affiliations

  1. Graduate Institute of Pharmaceutical Chemistry, College of Pharmacy, China Medical University, No 91, Hsueh-Shih Road, Taichung, 404, Taiwan

    Li-Chen Chou, Li-Jiau Huang & Sheng-Chu Kuo

  2. Department of Pharmacology, School of Medicine, College of Medicine, China Medical University, Taichung, 404, Taiwan

    Jai-Sing Yang

  3. School of Medicine, College of Medicine, China Medical University, Taichung, 404, Taiwan

    Hsi-Chin Wu & Kuan-Tin Chen

  4. Department of Life Sciences, College of Life Sciences, National Chung Hsing University, Taichung, 402, Taiwan

    Chi-Cheng Lu & Jo-Hua Chiang

  5. Department of Biological Science and Technology, College of Life Sciences, China Medical University, No 91, Hsueh-Shih Road, Taichung, 404, Taiwan

    Jing-Gung Chung

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  1. Li-Chen Chou
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  2. Jai-Sing Yang
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  7. Kuan-Tin Chen
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  8. Sheng-Chu Kuo
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Corresponding authors

Correspondence to Sheng-Chu Kuo or Jing-Gung Chung.

Additional information

L.-C. Chou and J.-S. Yang contributed equally to this work.

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Chou, LC., Yang, JS., Huang, LJ. et al. The synthesized 2-(2-fluorophenyl)-6,7-methylenedioxyquinolin-4-one (CHM-1) promoted G2/M arrest through inhibition of CDK1 and induced apoptosis through the mitochondrial-dependent pathway in CT-26 murine colorectal adenocarcinoma cells. J Gastroenterol 44, 1055–1063 (2009). https://doi.org/10.1007/s00535-009-0111-1

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  • DOI: https://doi.org/10.1007/s00535-009-0111-1

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