Alkaloid and acetogenin-rich fraction fromAnnona crassiflorafruit peel inhibits proliferation and migration of human liver cancer HepG2 cells

Abstract

Plant species from Annonaceae are commonly used in traditional medicine to treat various cancer types. This study aimed to investigate the antiproliferative potential of an alkaloid and acetogenin-rich fraction from the fruit peel of Annona crassiflora in HepG2 cells. A liquid-liquid fractionation was carried out on the ethanol extract of A. crassiflora fruit peel in order to obtain an alkaloid and acetogenin-rich fraction (AF-Ac). Cytotoxicity, proliferation and migration were evaluated in the HepG2 cells, as well as the proliferating cell nuclear antigen (PCNA), vinculin and epidermal growth factor receptor (EGFR) expression. In addition, intracellular Ca2+ was determined using Fluo4-AM and fluorescence microscopy. First, 9 aporphine alkaloids and 4 acetogenins that had not yet been identified in the fruit peel of A. crassiflora were found in AF-Ac. The treatment with 50 µg/mL AF-Ac reduced HepG2 cell viability, proliferation and migration (p < 0.001), which is in accordance with the reduced expression of PCNA and EGFR levels (p < 0.05). Furthermore, AF-Ac increased intracellular Ca2+ in the HepG2 cells, mobilizing intracellular calcium stores, which might be involved in the anti-migration and anti-proliferation capacities of AF-Ac. Our results support the growth-inhibitory potential of AF-Ac on HepG2 cells and suggest that this effect is triggered, at least in part, by PCNA and EGFR modulation and mobilization of intracellular Ca2+. This study showed biological activities not yet described for A. crassiflora fruit peel, which provide new possibilities for further in vivo studies to assess the antitumoral potential of A. crassiflora, especially its fruit peel.

Comprehensive Chemical Profiling and Mechanistic Insight into Anticancer Activity of Annona muricata Leaves Extract

“…The six phenolic compounds include sanguisorbic acid dilactone [9], galloyl-quinic acid-rutinoside [10], mucic acid-kaempferol-malic acid-rhamnose [10], galloyl pyrogallol derivative [10], digallic acid derivative [10], catechin derivative, and caffeic acid derivative [11] (Tables 1 and S2, Figure S1), and the six flavonoids include cyanidin-acetylglucoside pyruvic acid [12], chrysoeriol-7-O-glucouronyl-glucouronic acid [13], quercetin-rhmnosesophoroside [4], kaempferol-3-O-glucose-rhamnose-glucoside [4], tangeretin derivative [14], and visdulin III derivative [15] (Tables 1 and S3, Figure S1). Four alkaloids, squamolone [8], isopiline [16], vinblastine [17], and stepharine [4,14]), are also included (Tables 1 and S4, Figure S1), along with one lignan (phillygenin-O-hexose-O-pentose) [18] and one triterpenoid (2,3,19,23-tetra-hydroxy-urs-12-en-28-oic-acid-glucose [19]) (Tables 1 and S5, Figure S1). Conversely, the aqueous leaf capsule extract did not include the four acetogenins, named muridienin-1, muridienin-3, muridienin-4, and muricadienin, that we had previously identified (Table S1) [6].…”