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

Abstract

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.

In Vitro Cytotoxic Activity Test of Tampa Badak (Voacanga foetida (Blume) Leaves on L1210 Leukemia

“…Ekstrak etanol daun V. foetida memiliki efek analgesik pada dosis 250 mg/kg BB [8], ekstrak etil asetat dari daun V. foetida memiliki aktivitas sitotoksik pada sel kanker usus (HTB-38) dengan nilai IC 50 sebesar 0,1 g/mL [9], senyawa isolat Tb3 dari daun V. foetida memiliki efek antiproliferasi pada kanker leukemia (K562), paru (A549) dan serviks (He-La) dengan nilai IC 50 berturut-turut adalah 0,5; 2,4 dan 3,6 g/mL dibandingkan doksorubisin sebagai kontrol positif dengan nilai IC 50 0,2 g/mL [10], dan senyawa hasil isolasi (vobtusin) dari kulit batang tumbuhan V. foetida pada dosis 20 µM dapat menginduksi apoptosis melalui jalur mitokondria dengan cara mengaktivasi caspase-3 dan caspase-9 pada sel kanker leukemia HL-60 pada [2].…”

“…Salah satu alternatif dalam mencapai tujuan diatas dapat dilakukan dengan cara meneliti aktivitas sitotoksik dari tumbuhan, dan salah satu tumbuhan di Indonesia yang berpotensi sebagai calon obat antikanker baru adalah tumbuhan Voacanga foetida (Blume) Rolfe, termasuk famili Apocynaceae. Tumbuhan V. foetida mengandung metabolit sekunder alkaloid indole dan bisindole, dimana kedua alkaloid ini diketahui memiliki aktivitas sitotoksik [2]. Tumbuhan V. foetida yang dikenal dengan nama Tampa Badak, banyak ditemukan di provinsi Sumatera Barat, Riau, dan Jawa Barat, Jawa Tengah, dan Lombok.…”

Isolation and Cytotoxic Activity Test of Alkaloids from Dichloromethane Fraction of Bark of Tampa Badak (Voacanga foetida (Blume) Rolfe) Against T47D Cell Line

“…Estimated more than 2000 compounds classified as alkaloids [2]. Two bis-indole alkaloids from Voacanga foetida (Blume) Rolfe have been reported to have cytotoxic activity against HL60 [3] and A549 cell lines [4]. Alkaloid compounds as anticancer agents work by inducing apoptosis, inhibiting metastasis, inhibiting the cell cycle, inducing autophagy, inhibiting angiogenesis, inducing necrosis, activating mitochondrial pathway, and playing a role in tumor development [5].…”

In Vivo Cytotoxic Activity and Acute Toxicity Test of Ethanol Extract from Voacanga foetida (Blume) Rolfe Leaves

“…The Voacanga foetida (Apocynaceae) plant contains alkaloids such as voacangine, vobtusine, and vobtusine lactone has cytotoxic activity while vobtusine induces apoptosis via the mitochondria pathway [2].…”

“…Pharmacological studies have been carried out on V. foetida. According to these researchers, we know this plant has several pharmacological activities: analgesic [3], cytotoxic against colon cancer [4], antiproliferation on leukemia, lung, and cervical cancer [5], induces apoptosis, and inhibits the cell cycle in Leukemia cancer [2], and antimicrobial activity [6].…”