b-Mangostin induces p53-dependent G2/M cell cycle arrest and apoptosis through ROS mediated mitochondrial pathway and NfkB suppression in MCF-7 cells Suvitha Syam a,d, * , Ahmad Bustamam a, * , Rasedee Abdullah b , Mohamed Aspollah Sukari c , Najihah Mohd Hashim d , Mostafa Ghaderian d , Mawardi Rahmani c , Syam Mohan e , Siddig Ibrahim Abdelwahab e , Hapipah Mohd Ali f a UPM-MAKNA Cancer Research Laboratory, Institute of Bioscience, University Putra Malaysia, Serdang, Selangor, Malaysia b Department of Veterinary Pathology and Microbiology, Faculty of Veterinary, University Putra Malaysia, Serdang, Selangor, Malaysia c Department of Chemistry, Faculty of Science, University Putra Malaysia, Serdang, Selangor, Malaysia d Department of Pharmacy, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia e Medical Research Centre, Jazan University, P.O. Box 114, Jazan, Kingdom of Saudi Arabia f Department of Chemistry, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia ARTICLE INFO Article history: Received 10 July 2013 Received in revised form 23 October 2013 Accepted 24 October 2013 Available online xxxx Keywords: Cratoxylum arborescens b-Mangostin Apoptosis p53 NF-kB Bax/Bcl-2 ABSTRACT b-Mangostin (bM) was isolated from Cratoxylum arborescens to investigate its anti-cancer effect in MCF-7 cells. bM induced apoptosis by down-regulation of Bcl2 and up-regulation of Bax, triggering the cytochrome c release from mitochondria to cytosol. The release of caspase-9 and -7 and consequently cleaved PARP leading to apoptotic was observed upon treatment. Reduction of both bid and caspase 8 and the up regulation of Fas showed the involvement of the extrinsic pathway. Significantly up regulated GADD45A and HRK genes were observed upon treatment, with concomitant inhibition of NF-kB to nucleus. The pro- tein array had demonstrated the expression of HSP 70, HSP 60, XIAP, Survivin, p53 and Bax. Moreover, bM had showed p53-dependent G2/M cell cycle arrest by down regulation of cdc2 and PCNA. Together, the results demonstrated that the bM induced anti-proliferative effect, leading to G2/M phase cell cycle arrest and apoptosis through both the extrinsic and mito- chondrial pathways with the involvement of the multiple pro and anti-apoptosis and NF-kB signalling pathways. Ó 2013 Elsevier Ltd. All rights reserved. 1756-4646/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.jff.2013.10.018 * Corresponding authors. Address: UPM-MAKNA Cancer Research Laboratory, Institute of Bioscience, University Putra Malaysia, Serdang, Selangor, Malaysia (S. Syam). Tel.: +60 389472120; fax: +60 389472101. E-mail address: [email protected](S. Syam). Abbreviations: bM, b-mangostin; MMP, mitochondrial membrane potential; NF-kB, nuclear factor-kappa B; TNF-a, tumour necrosis factor alpha; ROS, reactive oxygen species; DCFH-DA, 2 0 ,7 0 -dichlorofluorescein diacetate; HCS, high content screening; PARP, poly ADP ribose polymerase; GADD, growth arrest DNA damage; XIAP, X-linked inhibitor of apoptosis protein; PMSF, phenylmethanesulfonylfluoride JOURNAL OF FUNCTIONAL FOODS xxx (2013) xxx – xxx Available at www.sciencedirect.com ScienceDirect journal homepage: www.elsevier.com/locate/jff Please cite this article in press as: Syam, S. et al., b-Mangostin induces p53-dependent G2/M cell cycle arrest and apoptosis through ROS mediated mitochondrial pathway and NfkB suppression in MCF-7 cells, Journal of Functional Foods (2013), http://dx.doi.org/10.1016/j.jff.2013.10.018
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J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 3 ) x x x – x x x
.sc ienced i rec t .com
Avai lab le a t www
ScienceDirect
journal homepage: www.elsevier .com/ locate / j f f
b-Mangostin induces p53-dependent G2/M cell cyclearrest and apoptosis through ROS mediatedmitochondrial pathway and NfkB suppressionin MCF-7 cells
1756-4646/$ - see front matter � 2013 Elsevier Ltd. All rights reserved.http://dx.doi.org/10.1016/j.jff.2013.10.018
* Corresponding authors. Address: UPM-MAKNA Cancer Research Laboratory, Institute of Bioscience, University Putra MalaysiaSelangor, Malaysia (S. Syam). Tel.: +60 389472120; fax: +60 389472101.
factor alpha; ROS, reactive oxygen species; DCFH-DA, 2 0,7 0-dichlorofluorescein diacetate; HCS, high content screening; PARPribose polymerase; GADD, growth arrest DNA damage; XIAP, X-linked inhibitor of apoptosis proteiphenylmethanesulfonylfluoride
Please cite this article in press as: Syam, S. et al., b-Mangostin induces p53-dependent G2/M cell cycle arrest and apoptosis through ROSmitochondrial pathway and NfkB suppression in MCF-7 cells, Journal of Functional Foods (2013), http://dx.doi.org/10.1016/j.jff.201
Suvitha Syama,d,*, Ahmad Bustamama,*, Rasedee Abdullahb, Mohamed Aspollah Sukaric,Najihah Mohd Hashimd, Mostafa Ghaderiand, Mawardi Rahmanic, Syam Mohane,Siddig Ibrahim Abdelwahabe, Hapipah Mohd Alif
aUPM-MAKNA Cancer Research Laboratory, Institute of Bioscience, University Putra Malaysia, Serdang, Selangor, MalaysiabDepartment of Veterinary Pathology and Microbiology, Faculty of Veterinary, University Putra Malaysia, Serdang, Selangor, MalaysiacDepartment of Chemistry, Faculty of Science, University Putra Malaysia, Serdang, Selangor, MalaysiadDepartment of Pharmacy, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, MalaysiaeMedical Research Centre, Jazan University, P.O. Box 114, Jazan, Kingdom of Saudi ArabiafDepartment of Chemistry, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia
A R T I C L E I N F O A B S T R A C T
Article history:
Received 10 July 2013
Received in revised form
23 October 2013
Accepted 24 October 2013
Available online xxxx
Keywords:
Cratoxylum arborescens
b-Mangostin
Apoptosis
p53
NF-kB
Bax/Bcl-2
b-Mangostin (bM) was isolated from Cratoxylum arborescens to investigate its anti-cancer
effect in MCF-7 cells. bM induced apoptosis by down-regulation of Bcl2 and up-regulation
of Bax, triggering the cytochrome c release from mitochondria to cytosol. The release of
caspase-9 and -7 and consequently cleaved PARP leading to apoptotic was observed upon
treatment. Reduction of both bid and caspase 8 and the up regulation of Fas showed the
involvement of the extrinsic pathway. Significantly up regulated GADD45A and HRK genes
were observed upon treatment, with concomitant inhibition of NF-kB to nucleus. The pro-
tein array had demonstrated the expression of HSP 70, HSP 60, XIAP, Survivin, p53 and Bax.
Moreover, bM had showed p53-dependent G2/M cell cycle arrest by down regulation of cdc2
and PCNA. Together, the results demonstrated that the bM induced anti-proliferative effect,
leading to G2/M phase cell cycle arrest and apoptosis through both the extrinsic and mito-
chondrial pathways with the involvement of the multiple pro and anti-apoptosis and NF-kB
Fig. 3 – Effect of bM on MMP, permeability and cytochrome c release. (A) Representative images of MCF-7 cells treated with
medium alone and 7 lg/ml of bM, and stained with Hoechst for nuclear, cell permeability dye, MMP and cytochrome c. The
images from each row were obtained from the same field of each sample (magnification 20·). (B–E) Average fluorescence
intensities of Hoechst 33342, cell permeability dye, MMP and cytochrome c in MCF-7 cells treated with bM or standard drug
Tamoxifen. Data were mean ± SD of fluorescence intensity readings measured from different photos taken (*P < 0.05).
8 J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 3 ) x x x – x x x
Please cite this article in press as: Syam, S. et al., b-Mangostin induces p53-dependent G2/M cell cycle arrest and apoptosis through ROS mediatedmitochondrial pathway and NfkB suppression in MCF-7 cells, Journal of Functional Foods (2013), http://dx.doi.org/10.1016/j.jff.2013.10.018
14 J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 3 ) x x x – x x x
MOHE/SC/09) from the Ministry of Higher Education Malaysia
and Makna Cancer research lab, UPM. The authors would like
to express their utmost gratitude to Late Dato Prof. Hamid A.
Hadi for his support during the research.
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