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Involvement of NF-κB and HSP70 signaling pathways in the apoptosis of MDA-MB-231 cells induced by a prenylated xanthone compound, α-mangostin, from Cratoxylum arborescens

Ibrahim MY 1 , Mohd Hashim N 1 , Mohan S 2 , Abdulla MA 3 , Abdelwahab SI 2 , Kamalidehghan B 1 Show all authors , Ghaderian M 4 , Dehghan F 5 , Ali LZ 1 , Karimian H 1 , Yahayu M 6 , Ee GC 7 , Farjam AS 8 , Mohd Ali H 9

Affiliations 

  • 1 Department of Pharmacy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia
  • 2 Medical Research Centre, Jazan University, Jazan, Saudi Arabia
  • 3 Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia
  • 4 Department of Pharmacy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia ; Epigenetics Lab, HIR Building, Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia
  • 5 Department of Pharmacy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia ; Department of Physiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia
  • 6 Department of Bioproduct Research and Innovation, Institute of Bioproduct Development (IBD), Universiti Teknologi Malaysia, UTM Johor Bahru, Johor, Malaysia
  • 7 Department of Chemistry, Faculty of Science, Universiti Putra Malaysia (UPM), Serdang, Selangor, Malaysia
  • 8 Institute of Tropical Agriculture, Universiti Putra Malaysia (UPM), Serdang, Selangor, Malaysia
  • 9 Department of Chemistry, University of Malaya, Kuala Lumpur, Malaysia
Drug Des Devel Ther, 2014;8:2193-211.
PMID: 25395836 DOI: 10.2147/DDDT.S66574

Abstract

BACKGROUND: Cratoxylum arborescens has been used traditionally in Malaysia for the treatment of various ailments.

METHODS: α-Mangostin (AM) was isolated from C. arborescens and its cell death mechanism was investigated. AM-induced cytotoxicity was observed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Acridine orange/propidium iodide staining and annexin V were used to detect cells in early phases of apoptosis. High-content screening was used to observe the nuclear condensation, cell permeability, mitochondrial membrane potential, and cytochrome c release. The role of caspases-3/7, -8, and -9, reactive oxygen species, Bcl-2 and Bax expression, and cell cycle arrest were also investigated. To determine the role of the central apoptosis-related proteins, a protein array followed by immunoblot analysis was conducted. Moreover, the involvement of nuclear factor-kappa B (NF-κB) was also analyzed.

RESULTS: Apoptosis was confirmed by the apoptotic cells stained with annexin V and increase in chromatin condensation in nucleus. Treatment of cells with AM promoted cell death-transducing signals that reduced MMP by downregulation of Bcl-2 and upregulation of Bax, triggering cytochrome c release from the mitochondria to the cytosol. The released cytochrome c triggered the activation of caspase-9 followed by the executioner caspase-3/7 and then cleaved the PARP protein. Increase of caspase-8 showed the involvement of extrinsic pathway. AM treatment significantly arrested the cells at the S phase (P<0.05) concomitant with an increase in reactive oxygen species. The protein array and Western blotting demonstrated the expression of HSP70. Moreover, AM significantly blocked the induced translocation of NF-κB from cytoplasm to nucleus.

CONCLUSION: Together, the results demonstrate that the AM isolated from C. arborescens inhibited the proliferation of MDA-MB-231 cells, leading to cell cycle arrest and programmed cell death, which was suggested to occur through both the extrinsic and intrinsic apoptosis pathways with involvement of the NF-κB and HSP70 signaling pathways.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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