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Ginsenoside Rh2-mediated G1 Phase Cell Cycle Arrest in Human Breast Cancer Cells Is Caused by p15Ink4B and p27Kip1-dependent Inhibition of Cyclin-dependent Kinases

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ABSTRACT

Purpose

Present study was undertaken to gain insights into the mechanism of cell cycle arrest by ginseng saponin ginsenoside Rh2 (Rh2) using MCF-7 and MDA-MB-231 breast cancer cells.

Methods

Cell viability and cell cycle distribution were determined by trypan blue dye exclusion assay and flow cytometry, respectively. Immunoblotting was performed to determine changes in protein levels. Knockdown of desired protein was achieved by transfection with small interfering RNA (siRNA).

Results

Rh2 treatment significantly inhibited viability of both cells in a concentration-dependent manner, which correlated with G0/G1 phase cell cycle arrest. Rh2-mediated cell cycle arrest was accompanied by down-regulation of cyclin-dependent kinases (Cdk) and cyclins leading to decreased interaction between cyclin D1 and Cdk4/Cdk6 and increased recruitment of p15Ink4B and p27Kip1 to cyclin D1/Cdk4 and cyclin D1/Cdk6 complexes. In addition, Rh2 treatment markedly reduced the levels of phosphorylated retinoblastoma protein (P-Rb) and decreased transcriptional activity of E2F1 in luciferase reporter assay. Rh2-induced cell cycle arrest was significantly attenuated by knockdown of p15Ink4B and/or p27Kip1 proteins.

Conclusions

Rh2-mediated cell cycle arrest in human breast cancer cells is caused by p15Ink4B and p27Kip1-dependent inhibition of kinase activities of G1-S specific Cdks/cyclin complexes.

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Authors and Affiliations

  1. Department of Life Science, Hallym University, Gangwon-do, Korea

    Sunga Choi

  2. Department of Biochemistry, Kangwon National University, Gangwon-do, Korea

    Tae Woong Kim

  3. Department of Pharmacology & Chemical Biology and University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    Shivendra V. Singh

Authors
  1. Sunga Choi
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  2. Tae Woong Kim
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  3. Shivendra V. Singh
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Corresponding authors

Correspondence to Sunga Choi or Shivendra V. Singh.

Additional information

Grant Support: This investigation was supported in part by the Kangwon Bio-Nuri grant, Korea Research Foundation, Korea, and US PHS grant CA129347, awarded by the National Cancer Institute.

Requests for reprints: S. Choi, Ph.D., Dept. of Life Science, Hallym University, 39 Hallymdaehak-gil, Chuncheon, Gangwon-do, 200-702, Korea. Phone: 82-33-248-2097; E-mail: sachoi@hallym.ac.kr or Shivendra V. Singh, 2.32A Hillman Cancer Center Research Pavilion, 5117 Centre Avenue, Pittsburgh, PA 15213. Phone: 412-623-3263; Fax: 412-623-7828. Email: singhs@upmc.edu

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Choi, S., Kim, T. & Singh, S.V. Ginsenoside Rh2-mediated G1 Phase Cell Cycle Arrest in Human Breast Cancer Cells Is Caused by p15Ink4B and p27Kip1-dependent Inhibition of Cyclin-dependent Kinases. Pharm Res 26, 2280–2288 (2009). https://doi.org/10.1007/s11095-009-9944-9

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  • DOI: https://doi.org/10.1007/s11095-009-9944-9

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