Nardostachys chinensis induces granulocytic differentiation with the suppression of cell growth through p27(Kip1) protein-related G0/G1 phase arrest in human promyelocytic leukemic cells

Pharm Biol. 2015 Jul;53(7):1002-9. doi: 10.3109/13880209.2014.952834. Epub 2014 Nov 28.

Abstract

Context: Nardostachys chinensis Batalin (Valerianaceae) has been used in Korean traditional medicine to elicit stomachic and sedative effects. However, the anti-leukemic activities of N. chinensis have not been well examined.

Objective: To investigate the effect of N. chinensis on differentiation and proliferation in the human promyelocytic leukemic HL-60 cells.

Materials and methods: The dried roots and stems of N. chiensis are extracted using hot water and then freeze-dried. The yield of extract was 12.82% (w/w). The HL-60 cells were treated with 25-200 μg/ml of N. chinensis for 72 h or 100 μg/ml of N. chinensis for 24-72 h.

Results: Nardostachys chinensis significantly inhibited cell viability dose dependently with an IC50 of 100 μg/ml in HL-60 cells. Nardostachys chinensis induced differentiation of the cells as measured by reduction activity of NBT and expression of CD11b but not of CD14 as analyzed by flow cytometry, which indicates a differentiation toward the granulocytic lineage. Nardostachys chinensis also induced growth inhibition through G0/G1 phase arrest in the cell cycle of HL-60 cells. Among the G0/G1 phase in the cell cycle-related protein, the expression of cyclin-dependent kinase (CDK) inhibitor p27(Kip1) was increased in N. chinensis-treated HL-60 cells, whereas the expression levels of CDK2, CDK4, CDK6, cyclin D1, cyclin D3, cyclin E, and cyclin A were decreased. Interestingly, N. chinensis markedly enhanced the binding of p27(Kip1) with CDK2 and CDK6.

Discussion and conclusion: This study demonstrated that N. chinensis is capable of inducing cellular differentiation and growth inhibition through p27(Kip1) protein-related G0/G1 phase arrest in HL-60 cells.

Keywords: Anti-leukemic effect; HL-60 cells; cell-cycle arrest; nardosinone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology
  • Cyclin-Dependent Kinase Inhibitor p27 / biosynthesis*
  • G1 Phase / drug effects*
  • G1 Phase / physiology
  • Granulocytes / drug effects*
  • Granulocytes / metabolism
  • Growth Inhibitors / isolation & purification
  • Growth Inhibitors / pharmacology*
  • HL-60 Cells
  • Humans
  • Leukemia, Promyelocytic, Acute / metabolism
  • Nardostachys*
  • Plant Extracts / isolation & purification
  • Plant Extracts / pharmacology*
  • Plant Roots
  • Plant Stems
  • Resting Phase, Cell Cycle / drug effects*
  • Resting Phase, Cell Cycle / physiology

Substances

  • CDKN1B protein, human
  • Growth Inhibitors
  • Plant Extracts
  • Cyclin-Dependent Kinase Inhibitor p27