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Cyclin D1 degradation and p21 induction contribute to growth inhibition of colorectal cancer cells induced by epigallocatechin-3-gallate

Abstract

Purpose

The public has paid attention to green tea due to its health benefits. Epigallocatechin-3-gallate (EGCG), the major component of green tea, is well documented to induce apoptosis and cell cycle arrest in cancer cells by targeting multiple signal transduction pathways. However, the detailed mechanism(s) of action needs to be determined.

Methods

Cell growth was evaluated by MTT assay, cell cycle analysis, and caspase 3/7 activity. Protein expression was analyzed through Western blotting. Reverse transcription polymerase chain reaction was used for examining mRNA expression of p21 and cyclin D1. The promoter activity of p21 was assessed by the luciferase reporter system.

Results

We identified cyclin D1 and p21 as molecular targets of EGCG in human colorectal cancer cells. We observed that cyclin D1 was down-regulated, while p21 expression was up-regulated by EGCG in dose- and time-dependent manners. Furthermore, we found EGCG decreased cyclin D1 protein stability, therefore triggering ubiquitin-dependent proteasomal degradation. Meanwhile, EGCG increased p21 promoter activity, resulting in up-regulation of p21 mRNA and protein, which was likely dependent on extracellular-signal-regulated kinase (ERK), inhibitor of nuclear factor kappa-B kinase (IKK) and phosphoinositide 3-kinase (PI3 K).

Conclusion

The data presented here details a novel mechanism by which EGCG inhibits cell growth of colorectal cancer cells. Namely, EGCG-induced cyclin D1 degradation and p21 transcriptional activation partially contribute to growth suppression in these cells.

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Abbreviations

EGCG:

Epigallocatechin-3-gallate

ECG:

Epicatechin gallate

EGC:

Epigallocatechin

EC:

Epicatechin

DMSO:

Dimethyl sulfoxide

FBS:

Fetal bovine serum

BSA:

Bovine serum albumin

PBS:

Phosphate-buffered saline

TBS:

Tris-buffered saline

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Acknowledgments

We thank Misty Bailey (University of Tennessee) for her critical reading of this manuscript. We also thank Drs. Kiyoshi Yamaguchi and Nichelle C. Whitlock for their technical assistance. This work was supported by NIH grant R01CA108975, and The University of Tennessee Center of Excellence in Livestock Diseases and Human Health grant to SJB. Financial support for XZ was provided by the Program in Organizational or Personal Cooperation with Foreign Counterparts (2010630161), China Scholarship Council, China.

Conflict of interest

None.

Author information

Correspondence to Seung Joon Baek.

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Zhang, X., Min, K., Wimalasena, J. et al. Cyclin D1 degradation and p21 induction contribute to growth inhibition of colorectal cancer cells induced by epigallocatechin-3-gallate. J Cancer Res Clin Oncol 138, 2051–2060 (2012). https://doi.org/10.1007/s00432-012-1276-1

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Keywords

  • EGCG
  • Cyclin D1
  • Proteasome
  • p21
  • Colorectal cancer