Vobtusine from Voacanga foetida (Blume) Rolfe Induces Apoptosis via Activation of Caspase Pathway in Human HL-60 Leukemia Cell Line

Adriani Susanty, - Dachriyanus, - Yanwirasti, Fatma Sri Wahyuni, Puteri Amelia, Saori Miyama, Yusuke Hirasawa, Toshio Kaneda, Hiroshi Morita


Vobtusine is an aspidosperma-aspidosperma alkaloid isolated from alkaloid DCM base fraction of the bark of Voacanga foetida (Blume) Rolfe (Apocynaceae). In this study, the effect of vobtusine on the cell cycle, apoptosis induction, and Bcl-2 family protein expression  were investigated by flow cytometr,  DNA fragmentation analysis, and western blotting, respectively. The results of cell cycle analysis indicated the ratio of the number of cells in each phase did not have significant differences depend on vobtusine concentration although cell number in G1 phase had tendency to decrease according to the increasing of vobtusine concentration. Besides, the sub-G1 phase population of HL-60 cells treated with vobtusine was increased compared with that of cells without treatments (5.9%-23.8%). DNA fragmentation was observed from 20 µM, and the degree of fragmentation was dependent on vobtusine concentration. Caspase-3 activity increased 4.6 times compared to control. After being treated with caspase-9 inhibitor, vobtusine-induced elevation of caspase-3 activity decreased. This shows that caspase-3 activity depends on caspase-9. Vobtusine was also induced apoptotic cell death involved a mitochondrial by Bid activation and Bcl-xL downregulation. Therefore, vobtsusine-induced apoptosis process was initiated by caspase-9 via change of Bcl-2 family protein expression and executed by caspase-3, followed by cell death due to their proteolytic activity. These results indicated the mechanism action of vobtusine as anti-cancer compound via intrinsic pathway.


vobtusine; cell cycle, apoptosis; caspases (-8, -9, -3); Bid; Bcl-xL.

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F. J. Lipp, L. M. Perry, and J. Metzger, “Medicinal Plants of East and Southeast Asia: Attributed Properties and Uses.,†Brittonia, 1981.

H. Almehdar, H. M. Abdallah, A. M. M. Osman, and E. A. Abdel-Sattar, “In vitro cytotoxic screening of selected Saudi medicinal plants,†J. Nat. Med., 2012.

B. S. Bhadane, M. P. Patil, V. L. Maheshwari, and R. H. Patil, “Ethnopharmacology, phytochemistry, and biotechnological advances of family Apocynaceae: A review,†Phytotherapy Research. 2018.

S. Hadi, B. Desy Ratnasari, M. Septiyana, S. Priyambodo, and I. M. Sudarma, “Antibacterial Assay and Alkaloid Lombine Distribution Study of Voacanga foetida (B.I) Rolfe from Lombok Island,†Orient. J. Chem., 2019.

Y. Tang et al., “Leucophyllinines A and B, bisindole alkaloids from Leuconotis eugeniifolia,†J. Nat. Med., vol. 73, no. 3, pp. 533–540, 2019.

Prema et al., “Three new quassinoids isolated from the wood of Picrasma javanica and their anti-Vpr activities,†J. Nat. Med., vol. 74, no. 3, pp. 571–578,2020.

Y. Hirasawa et al., “New vasorelaxant indole alkaloids, taberniacins A and B, from Tabernaemontana divaricata,†J. Nat. Med., vol. 73, no. 3, pp. 627–632, 2019.

P. Amelia et al., “Two new sarpagine-type indole alkaloids and antimalarial activity of 16-demethoxycarbonylvoacamine from Tabernaemontana macrocarpa Jack,†J. Nat. Med., vol. 73, no. 4, pp. 820–825, 2019.

Dachriyanus, D. Arbain, D. P. Putra, M. V. Sargent, R. Susila, and F. S. Wahyuni, “Indole alkaloids from two species of Ophiorrhiza,†Aust. J. Chem., 2000.

Dachriyanus, M. V. Sargent, and F. S. Wahyuni, “(+)-Isochimonanthine, a pyrrolidinoindole alkaloid from Argostemma yappii king,†Aust. J. Chem., vol. 53, no. 2, pp. 159–160, 2000.

D. Arbain, A. L. T. Byrne, B. Dachriyanus, N. E. A, and M. V. S. B, “of Chemistry,†Aust. J. Chem., vol. 50, pp. 3–5, 1997.

D. Arbain, L. T. Byrne, Dachriyanus, and M. V. Sargent, “Isomalindine-16-carboxylate, a zwitterionic alkaloid from Ophiorrhiza cf. communis,†Aust. J. Chem., 1997.

A. Susanty, D. Dachriyanus, Y. Yanwirasti, F. S. Wahyuni, H. Fadhli, and P. A. Aswan, “Aktivitas Sitotoksik Ekstrak Etil Asetat Daun Tampa Badak (Voacanga foetida (Bl.)K.Schum) pada Kanker Kolon HTB-38,†JSFK (Jurnal Sains Farm. Klin., vol. 5, no. 2, pp. 142–146, 2018.

A. Burlacu, “Regulation of apoptosis by Bcl-2 family proteins,†Journal of Cellular and Molecular Medicine. 2003.

R. Jan and G. e. S. Chaudhry, “Understanding apoptosis and apoptotic pathways targeted cancer therapeutics,†Advanced Pharmaceutical Bulletin. 2019.

S. Elmore, “Apoptosis: A Review of Programmed Cell Death,†Toxicol. Pathol., vol. 35, no. 4, pp. 495–516, 2007.

P. Majtnerová and T. Roušar, “An overview of apoptosis assays detecting DNA fragmentation,†Mol. Biol. Rep., vol. 45, no. 5, pp. 1469–1478, 2018.

D. Daddiouaissa, A. Amid, N. A. Kabbashi, F. A. A. Fuad, A. A. M. Elnour, and M. A. K. M. S. Epandy, “Antiproliferative activity of ionic liquid-graviola fruit extract against human breast cancer (MCF-7) cell lines using flow cytometry techniques,†J. Ethnopharmacol., vol. 236, no. December 2018, pp. 466–473, 2019.

R. Singh, A. Letai, and K. Sarosiek, “Regulation of apoptosis in health and disease: the balancing act of BCL-2 family proteins,†Nat. Rev. Mol. Cell Biol., vol. 20, no. 3, pp. 175–193, 2019.

Z. Fu et al., “Euphorbia lunulata extract acts on multidrug resistant gastric cancer cells to inhibit cell proliferation, migration and invasion, arrest cell cycle progression, and induce apoptosis,†J. Ethnopharmacol., 2018.

K. Zaima et al., “Biscarpamontamines A and B, an aspidosperma-iboga bisindole alkaloid and an aspidosperma-aspidosperma bisindole alkaloid, from Tabernaemontana sphaerocarpa,†J. Nat. Prod., vol. 72, no. 9, pp. 1686–1690, 2009.

T. Kaneda, M. Matsumoto, Y. Sotozono, S. Fukami, and A. Eko, “Cycloartane triterpenoid ( 23 R , 24 E )‑ 23 ‑ acetoxymangiferonic acid inhibited proliferation and migration in B16 ‑ F10 melanoma via MITF downregulation caused by inhibition of both β ‑ catenin and c ‑ Raf – MEK1 – ERK signaling axis,†J. Nat. Med., vol. 74, no. 1, pp. 47–58, 2020.

DOI: http://dx.doi.org/10.18517/ijaseit.10.6.12353


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