Baicalin induces apoptosis in leukemia HL-60/ADR cells via possible down-regulation of the PI3K/Akt signaling pathway

Asian Pac J Cancer Prev. 2012;13(4):1119-24. doi: 10.7314/apjcp.2012.13.4.1119.

Abstract

Background: The effect and possible mechanism of traditional Chinese medicine, baicalin, on the PI3K/ Akt signaling pathway in drug-resistant human myeloid leukemia HL-60/ADR cells have been investigated in this current study.

Methods: HL-60/ADR cells were treated by 20, 40, 80 μmol/L baicalin followed by cell cycle analysis at 24h. The mRNA expression level of the apoptosis related gene, Bcl-2 and bad, were measured by RT-PCR on cells treated with 80 μmol/L baicalin at 12, 24 and 48hr. Western blot was performed to detect the changes in the expression of the proteins related to HL-60/ADR cell apoptosis and the signaling pathway before and after baicalin treatment, including Bcl-2, PARP, Bad, Caspase 3, Akt, p-Akt, NF-κB, p-NF-κB, mTOR and p-mTOR.

Results: Sub-G1 peak of HL-60/ADR cells appeared 24 h after 20 μmol/L baicalin treatment, and the ratio increased as baicalin concentration increased. Cell cycle analysis showed 44.9% G0/G1 phase cells 24 h after baicalin treatment compared to 39.6% in the control group. Cells treated with 80 μmol/L baicalin displayed a trend in decreasing of Bcl-2 mRNA expression over time. Expression level of the Bcl-2 and PARP proteins decreased significantly while that of the PARP, Caspase-3, and Bad proteins gradually increased. No significant difference in Akt expression was observed between treated and the control groups. However, the expression levels of p-Akt, NF-κB, p-NF-κB, mTOR and p-mTOR decreased significantly in a time-dependent manner.

Conclusions: We conclude that baicalin may induce HL-60/ADR cell apoptosis through the PI3K/AKT signaling pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects
  • Caspase 3 / drug effects
  • Caspase 3 / metabolism
  • Down-Regulation / drug effects
  • Drug Resistance, Neoplasm
  • Drugs, Chinese Herbal / pharmacology
  • Enzyme Inhibitors / pharmacology*
  • Flavonoids / pharmacology*
  • G1 Phase
  • Gene Expression
  • HL-60 Cells
  • Humans
  • NF-kappa B / drug effects
  • NF-kappa B / metabolism
  • Phosphatidylinositol 3-Kinases / drug effects
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphorylation / drug effects
  • Poly(ADP-ribose) Polymerases / drug effects
  • Poly(ADP-ribose) Polymerases / metabolism
  • Proto-Oncogene Proteins c-akt / drug effects
  • Proto-Oncogene Proteins c-akt / metabolism*
  • Proto-Oncogene Proteins c-bcl-2 / drug effects
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • RNA, Messenger / drug effects
  • RNA, Messenger / metabolism
  • Resting Phase, Cell Cycle
  • Signal Transduction / drug effects*
  • TOR Serine-Threonine Kinases / drug effects
  • TOR Serine-Threonine Kinases / metabolism
  • bcl-Associated Death Protein / drug effects
  • bcl-Associated Death Protein / genetics
  • bcl-Associated Death Protein / metabolism

Substances

  • BAD protein, human
  • Drugs, Chinese Herbal
  • Enzyme Inhibitors
  • Flavonoids
  • NF-kappa B
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Messenger
  • bcl-Associated Death Protein
  • baicalin
  • Poly(ADP-ribose) Polymerases
  • MTOR protein, human
  • Proto-Oncogene Proteins c-akt
  • TOR Serine-Threonine Kinases
  • Caspase 3