Planta Medica International Open 2017; 4(S 01): S1-S202
DOI: 10.1055/s-0037-1608325
Lecture Session – Bioactive Natural Products II
Georg Thieme Verlag KG Stuttgart · New York

Natural products upregulating Smac/DIABLO genes expression from Ficus deltoidea and their role in prostate cancer chemoprevention.

M Mohd Hanafi Mohd
1   Department of Pharmaceutical and Biological Chemistry, UCL School of Pharmacy, London, United Kingdom
,
A Afzan
2   School of Pharmaceutical Sciences, EPGL, University of Geneva, University of Lausanne, Geneva, Switzerland
,
H Yaakob
3   Institute of Bioproduct Development (IBD), Universiti Teknologi Malaysia, 81310 UTM, Johor Bahru, Malaysia
,
R Aziz
3   Institute of Bioproduct Development (IBD), Universiti Teknologi Malaysia, 81310 UTM, Johor Bahru, Malaysia
,
R Sarmidi Mohamad
3   Institute of Bioproduct Development (IBD), Universiti Teknologi Malaysia, 81310 UTM, Johor Bahru, Malaysia
,
JL Wolfender
2   School of Pharmaceutical Sciences, EPGL, University of Geneva, University of Lausanne, Geneva, Switzerland
,
M Prieto Jose
1   Department of Pharmaceutical and Biological Chemistry, UCL School of Pharmacy, London, United Kingdom
› Author Affiliations
Further Information

Publication History

Publication Date:
24 October 2017 (online)

 

This study aims to evaluate the in-vitro cytotoxic and anti-migratory effects of Ficus deltoidea L. plant on prostate cancer cells, identify the active compound/s and characterise their mechanism of actions. The crude methanolic extract was partitioned into n-hexane (FD1h, FD2h) chloroform (FD1c, FD2c) and aqueous extracts (FD1a, FD2a). Active fractions were further fractionated. Active compound/s were dereplicated based on UHPLC-HRMS/MS. In-vitro mechanistic studies on PC3 cells included: Annexin V-FITC staining, MMP depolarization measurements, activity of caspases 3/7, nuclear DNA fragmentation, cell cycle analysis, modulation of Bax, Bcl-2, Smac/Diablo and Alox-5 mRNA gene expression by RT-PCR. 2D migration and 3D invasion were tested by exclusion assays and modified Boyden chamber [1]. The expression of VEGF-A, CXCR4 and CXCL12 in PC3 cells were analysed by RT-PCR. FD1c and FD2c extracts induced cell death (P < 0.05) via apoptosis as evidenced by nuclear DNA fragmentation accompanied by an increase in MMP depolarization (P < 0.05), and activation of caspases 3/7 (P < 0.05). All active plant extracts up-regulated Bax and Smac/DIABLO, and down-regulated Bcl-2 (P < 0.05). Active plant extracts inhibited both migration and invasion of PC3 cells (P < 0.05). These effects were accompanied by down-regulation of both VEGF-A and CXCL-12 gene expressions (P < 0.001). UHPLC-HRMS/MS dereplication identified isovitexin in FD1c; and oleanolic acid, moretenol, betulin, lupenone and lupeol in FD2c. In conclusion, FD1c and FD2c were able to induce apoptosis by activating the intrinsic pathway, inhibit both migration and invasion by modulating the CXCL12-CXCR4 axis, and inhibit angiogenesis by modulating VEGF-A expression in PC3 cell line.

I would like to thank the Foundation Plants for Health for awarding me with a research subsidy and giving me the opportunity to collaborate with Prof Jean-Luc Wolfender in completing this project.

[1] Ridley, Anne J, et al. Science 302.5651 (2003): 1704 – 1709.