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(3′R)-hydroxytabernaelegantine C: A bisindole alkaloid with potent apoptosis inducing activity in colon (HCT116, SW620) and liver (HepG2) cancer cells

Author:
Paterna, Angela, Gomes, Sofia E., Borralho, Pedro M., Mulhovo, Silva, Rodrigues, Cecília M.P., Ferreira, Maria-José U.
Source:
Journal of ethnopharmacology 2016 v.194 pp. 236-244
ISSN:
0378-8741
Subject:
NAD ADP-ribosyltransferase, Tabernaemontana, alkaloids, apoptosis, cell viability, colon, colorectal neoplasms, cytotoxicity, fibroblasts, flow cytometry, fluorouracil, fractionation, guavas, human cell lines, humans, inhibitory concentration 50, lactate dehydrogenase, liver, medicinal plants, methanol, neoplasm cells, nuclear magnetic resonance spectroscopy, pro-apoptotic proteins, roots, staining, traditional medicine, Africa
Abstract:
Tabernaemontana elegans Stapf. (Apocynaceae) is a medicinal plant traditionally used in African countries to treat cancer.To discover new apoptosis inducing lead compounds from T. elegans and provide scientific validation of the ethnopharmacological use of this plant.Through fractionation, (3′R)-hydroxytaberanelegantine C (1), a vobasinyl-iboga bisindole alkaloid, was isolated from a cytotoxic alkaloid fraction of the methanol extract of T. elegans roots. Its structure was identified by spectroscopic methods, mainly 1D and 2D NMR experiments. Compound 1 was evaluated for its ability to induce apoptosis in HCT116 and SW620 colon and HepG2 liver carcinoma cells. The cell viability of compound 1 was evaluated by the MTS and lactate dehydrogenase (LDH) assays. Induction of apoptosis was analyzed through Guava ViaCount assay, by flow cytometry, caspase-3/7 activity assays and evaluation of nuclear morphology by Hoechst staining. To determine the molecular pathways elicited by 1 exposure, immunoblot analysis was also performed.(3′R)-hydroxytaberanelegantine C (1) displayed strong apoptosis induction activity as compared to 5-fluorouracil (5-FU), the most used anticancer agent in colorectal cancer treatment. In the MTS assay, compound 1 exhibited IC50 values similar or lower than 5-FU in the three cell lines tested. The IC50 value of 1 was also calculated in CCD18co normal human colon fibroblasts. The lactate dehydrogenase assay showed increased LDH release by compound 1, and the Guava ViaCount assay revealed that 1 significantly increased the incidence of apoptosis to a further extent than 5-FU. Moreover, the induction of apoptosis was corroborated by evaluation of nuclear morphology by Hoechst staining and caspase-3/7 activity assays of 1 treated cells. As expected, in immunoblot analysis, compound 1 treatment led to poly(ADP-ribose) polymerase cleavage. This was accompanied by decreased anti-apoptotic proteins Bcl-2 and XIAP steady state levels in all three cancer cell lines tested.Compound 1 showed remarkable induction of apoptosis in HCT116, SW620 and HepG2 cells. Together, the results suggest that compound 1 is a promising lead structure for inducing apoptosis.
Agid:
5598266