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Apoptosis

, Volume 12, Issue 10, pp 1911–1926 | Cite as

A triterpenediol from Boswellia serrata induces apoptosis through both the intrinsic and extrinsic apoptotic pathways in human leukemia HL-60 cells

  • Shashi Bhushan
  • Ajay Kumar
  • Fayaz Malik
  • Samar Singh Andotra
  • Vijay Kumar Sethi
  • Indu Pal Kaur
  • Subhash Chandra Taneja
  • Ghulam Nabi Qazi
  • Jaswant Singh
Original Paper

Abstract

A triterpenediol (TPD) comprising of isomeric mixture of 3α, 24-dihydroxyurs-12-ene and 3α, 24-dihydroxyolean-12-ene from Boswellia serrata induces apoptosis in cancer cells. An attempt was made in this study to investigate the mechanism of cell death by TPD in human leukemia HL-60 cells. It inhibited cell proliferation with IC50 ∼ 12 μg/ml and produced apoptosis as measured by various biological end points e.g. increased sub-G0 DNA fraction, DNA ladder formation, enhanced AnnexinV-FITC binding of the cells. Further, initial events involved massive reactive oxygen species (ROS) and nitric oxide (NO) formation, which were significantly inhibited by their respective inhibitors. Persistent high levels of NO and ROS caused Bcl-2 cleavage and translocation of Bax to mitochondria, which lead to loss of mitochondrial membrane potential (Δψm) and release of cytochrome c, AIF, Smac/DIABLO to the cytosol. These events were associated with decreased expression of survivin and ICAD with attendant activation of caspases leading to PARP cleavage. Furthermore, TPD up regulated the expression of cell death receptors DR4 and TNF-R1 level, leading to caspase-8 activation. These studies thus demonstrate that TPD produces oxidative stress in cancer cells that triggers self-demise by ROS and NO regulated activation of both the intrinsic and extrinsic signaling cascades.

Keywords

Boswellia serrata Triterpenediol Apoptosis HL-60 cells RNOS cytochrome c 

Abbreviations

DAF-2-DA

Diaminofluoresceine-2-diacetate

DCFH-DA

2,7-Dichlorofluoresceine diacetate

LPS

Lipopolysaccharide

Δψm

Mitochondrial membrane potential

MTT

3-(4,5-Dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide

PBMC

Peripheral blood mononuclear cell

NAC

N-acetyl-cysteine

iNOS

Inducible nitric oxide synthase

PI

Propidium iodide

PTP

Mitochondrial permeability transition pore

NO

Nitric oxide

Rh-123

Rhodamine-123

sMIT

s-Methylisothiourea

AIF

Apoptosis inducing factor

PMSF

Phenylmethanesulfonyl fluoride

ICAD

Inhibitor of caspase activating DNAse

Notes

Acknowledgements

We are highly thankful to Dr. Sarang Bani and Sheikh Fayaz (SRF, CSIR) for their help in the use of Flow Cytometery.

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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Shashi Bhushan
    • 1
  • Ajay Kumar
    • 1
  • Fayaz Malik
    • 1
  • Samar Singh Andotra
    • 1
  • Vijay Kumar Sethi
    • 1
  • Indu Pal Kaur
    • 2
  • Subhash Chandra Taneja
    • 1
  • Ghulam Nabi Qazi
    • 1
  • Jaswant Singh
    • 1
  1. 1.Division of PharmacologyIndian Institute of Integrative MedicineJammuIndia
  2. 2.University Institute of Pharmaceutical SciencesPanjab UniversityChandigarhIndia

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