Alkaloid and acetogenin-rich fraction from Annona crassiflora fruit 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.

Scientific Advances in the Last Decade on the Recovery, Characterization, and Functionality of Bioactive Compounds from the Araticum Fruit (Annona crassiflora Mart.)

“…The stephalagine was purified by semi-preparative HPLC on RP-C 18 and structurally characterized by high-resolution mass spectrometry, with electrospray ionization (HR-ESI-MS) and nuclear magnetic resonance (NMR) techniques, for the first time by Pereira et al [ 29 ]. Additionally, recent studies have reported the presence of stephalagine in araticum peel and pulp [ 14 , 26 , 27 , 28 , 30 ], indicating that this compound is present in more than a fraction of the fruit. Stephalagine, an aporphine alkaloid, has demonstrated anti-obesity, antinociceptive, and anti-edematogenic effects, suggesting its use as a new potential drug [ 28 , 29 ].…”

Bcl-2-like protein-10 increases aggressive features of melanoma cells

“…Thus, the role of Bcl-2 proteins in regulating intracellular and mitochondrial Ca 2+ homeostasis, and subsequently to several other processes such as cell migration, independently of apoptosis, is emerging [ 53 ]. Increased intracellular Ca 2+ levels have been described to be associated to the anti-migration effects of alkaloid compounds in liver cancer cells [ 54 ]. Based on this consideration, we can hypothesize that the increased migration properties observed in melanoma cells overexpressing Bcl2L10 could also be linked to the reduced [Ca 2+ ] i detected in the same cells.…”

Phytochemical screening and potential antioxidant, antibacterial and anti-melanoma activities of <i>Rollinia dolabripetala</i> leaves

“…In our experimental tests, analyses of phytocompounds such as alkaloids, present in many annonaceae (Chen et al, 2021; E.V. Costa et al, 2021;Justino et al, 2021;Kumar et al, 2021;Pandiyan & Ilango, 2022;Perrone et al, 2022;Rocha et al, 2021) that also exert antioxidant activity, were not included. In A. muricata, the strong antioxidant effect is highly correlated, mainly, to phytochemicals such as flavonoids and phenolic compounds, however, several other antioxidant compounds have been identified, such as tannins, saponins, glycoside, and acetogenins in this species (Esparza & González, 2020;Wahab et al, 2018) and other annonaceous (Andrade et al, 2021;Gowsalya & Sathya, 2021;Kumar et al, 2021;Perrone et al, 2022;Villalva et al, 2021).…”

“…Nascimento et al, 2003). Annonaceous acetogenins constitute a series of polyketides found almost exclusively in plants of the Annonaceae family with bioactivity and mechanisms of action of scientific interest (Justino et al, 2021;Kazman et al, 2022;R.D.P. Nascimento et al, 2022;Neske et al, 2020).…”

“…Leaf extracts of Annona crassiflora that had tannins (Ribeiro et al, 2018;Sieniawska & Baj, 2017;Smeriglio et al, 2017) and xanthone derivatives removed, no inhibitory effects on bacterial growth were observed (Ribeiro et al, 2018). Alkaloids, phytochemicals present in many annonaceae (Dev & Joseph, 2021;Justino et al, 2021;Perrone et al, 2022;Verma, 2020), also exert antibacterial action including Staphylococcus (Marco et al, 2019;Perrone et al, 2022). This is the first study of the antitumoral, antioxidant and antibacterial bioactivity of R. dolabripetala leaf extracts (RDLE).…”