Journal of Natural Medicines

, Volume 71, Issue 1, pp 36–43 | Cite as

AU-1 from Agavaceae plants causes transient increase in p21/Cip1 expression in renal adenocarcinoma ACHN cells in an miR-34-dependent manner

  • Tomofumi Fujino
  • Akihito Yokosuka
  • Hideaki Higurashi
  • Rina Yokokawa
  • Ryo Sakurai
  • Wataru Harashima
  • Yuichi Miki
  • Yasuyuki Fujiwara
  • Yoshihiro Mimaki
  • Makio Hayakawa
Original Paper

Abstract

Here, we show that AU-1, spirostanol saponin isolated from Agavaceae plants, causes a transient increase in cyclin-dependent kinase inhibitor (CDKI) p21/Cip1 through the upregulation of miRNAs, miR-34 and miR-21. AU-1 stimulated p21/Cip1 expression without exerting cytotoxicity against different types of carcinoma cell lines. In renal adenocarcinoma ACHN cells, AU-1 transiently elevated the expression level of p21/Cip1 protein without marked increases in p21/Cip1 mRNA levels. Rapid and transient increases in miR-34 and miR-21, both of which are known to upregulate p21/Cip1, were observed in AU-1-treated cells. Inhibitor for miR-34 and for miR-21 significantly blocked the AU-1-caused increase in p21/Cip1, indicating that elevation of p21/Cip1 protein by AU-1 is dependent on these microRNAs. We further clarified that NAD-dependent deacetylase SIRT1, a direct target of miR-34, is decreased by the treatment with AU-1. Furthermore, we found that SIRT1-knockdown increases p21/Cip1 protein levels in an miR-21-dependent manner. On the other hand, ectopic expression of p21/Cip1 resulted in the lowered expression of miR-34 and miR-21, suggesting that reciprocal regulation exists between p21/Cip1 and these miRNAs. We propose that the following feedback network composed of miR-34/SIRT1/miR-21/p21 is triggered by the treatment with AU-1: in cells treated with AU-1, transient elevation of miR-34 leads to the downregulation of SIRT1, thereby miR-21 is freed from SIRT1-dependent suppression. Then, elevated miR-21 upregulates p21/Cip1 protein, followed by the suppression of miR-34 expression.

Keywords

Agave utahensis p21/Cip1 miR-34 miR-21 SIRT1 

Notes

Acknowledgments

We thank Ken Ando, Toshiyuki Oshima, and Harutaka Ichikawa for their helpful advice and discussions. This work was supported, in part, by a grant from the Japan Private School Promotion Foundation.

Compliance with ethical standards

Conflict of interest

None declared.

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Copyright information

© The Japanese Society of Pharmacognosy and Springer Japan 2016

Authors and Affiliations

  • Tomofumi Fujino
    • 1
  • Akihito Yokosuka
    • 2
  • Hideaki Higurashi
    • 1
  • Rina Yokokawa
    • 1
  • Ryo Sakurai
    • 1
  • Wataru Harashima
    • 1
  • Yuichi Miki
    • 3
  • Yasuyuki Fujiwara
    • 3
  • Yoshihiro Mimaki
    • 2
  • Makio Hayakawa
    • 1
  1. 1.Department of Hygiene and Health Sciences, School of PharmacyTokyo University of Pharmacy and Life SciencesHachiōjiJapan
  2. 2.Department of Medicinal Pharmacognosy, School of PharmacyTokyo University of Pharmacy and Life SciencesHachiōjiJapan
  3. 3.Department of Environmental Health, School of PharmacyTokyo University of Pharmacy and Life SciencesHachiōjiJapan

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