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Molecular mechanisms of ginsenoside Rh2-mediated G1 growth arrest and apoptosis in human lung adenocarcinoma A549 cells

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Abstract

Ginsenoside Rh2 (Rh2), a purified ginseng saponin, has been shown to have antiproliferative effects in certain cancer cell types. However, the molecular mechanisms of Rh2 on cell growth and death have not been fully clarified. In this study, the antiproliferative effect of Rh2 in human lung adenocarcinoma A549 cells was investigated. Treatment of A549 cells with 30 μg/ml Rh2 resulted in G1 phase arrest, followed by progression to apoptosis. This Rh2-mediated G1 arrest was accompanied by downregulation of the protein levels and kinase activities of cyclin-D1, cyclin-E and Cdk6, and the upregulation of pRb2/p130. In addition, Rh2-induced apoptosis was confirmed by TUNEL assay and DNA fragmentation analysis. Administration of Rh2 caused an increase in the expression levels of TRAIL-RI (DR4) death receptor but did not alter the levels of other death receptors or Bcl-2 family molecules. Furthermore, the Rh2-induced apoptosis was significantly inhibited by DR4:Fc fusion protein, which inhibits TRAIL-DR4-mediated apoptosis. In addition, caspase-2, caspase-3 and caspase-8 were highly activated upon Rh2 treatment. Inhibitors of caspase-2, caspase-3 and caspase-8 markedly prevented the cell death induced by Rh2. Inhibitor of caspase-8 significantly inhibited the activation of caspase-2, caspase-3 and caspase-8. These observations indicate that multiple G1-related cell cycle regulatory proteins are regulated by Rh2 and contribute to Rh2-induced G1 growth arrest. The increase in the expression level of DR4 death receptor may play a critical role in the initiation of Rh2-triggered apoptosis, and the activation of the caspase-8/caspase-3 cascade acts as the executioner of the Rh2-induced death process.

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Abbreviations

Adv:

Adenovirus

Cdk:

Cycline-dependent kinase

Rb:

Retinoblastoma

Rh2:

Ginsenoside Rh2

TRAIL:

TNF-related apoptosis-inducing ligand

TUNEL:

Terminal transferase-mediated dUTP-fluorescein nick end-labelling

z-VDVAD-fmk:

z-Val-Asp-Val-Ala-Asp-fluoromethyl ketone

z-DEVD-fmk:

z-Asp-Glu-Val-Asp-fluoromethyl ketone

z-IETD-fmk:

z-Ile-Glu-Thr-Asp-fluoromethyl ketone

z-LEHD-fmk:

z-Leu-Glu-His-Asp-fluoromethyl ketone

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Acknowledgements

This work is supported by grants from the Taichung Veterans General Hospital TCVGH-927312D and the National Science Council NSC 91-2311-B075A-001, Taiwan, ROC.

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Authors and Affiliations

  1. Department of Education and Research, Taichung Veterans General Hospital, Taichung, 407, Taiwan

    Chi-Chih Cheng & Shih-Lan Hsu

  2. Institute of Chinese Medical Science, China Medical University, Taichung, Taiwan

    Shu-Mei Yang, Jung-Chou Chen & Shih-Lan Hsu

  3. Division of Molecular and Genomic Medicine, National Health Research Institutes, Taipei, Taiwan

    Chi-Ying Huang

  4. Savelife Bioscience Company, Taichung, Taiwan

    Wei-Mao Chang

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  1. Chi-Chih Cheng
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Correspondence to Shih-Lan Hsu.

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Cheng, CC., Yang, SM., Huang, CY. et al. Molecular mechanisms of ginsenoside Rh2-mediated G1 growth arrest and apoptosis in human lung adenocarcinoma A549 cells. Cancer Chemother Pharmacol 55, 531–540 (2005). https://doi.org/10.1007/s00280-004-0919-6

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