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Rita Sarkis
Department of Natural Sciences, Lebanese American University, Byblos, Lebanon

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Journal article
Published: 02 June 2021 in Biomedicine & Pharmacotherapy
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Annonaceae family has broad uses in herbal medicine for treatment of several diseases, whether through seeds’ or leaves’ extracts. The present study investigates the antiproliferative and antitumor activity of Annona cherimola aqueous leaf (AAL) extract/infusion in acute myeloid leukemia (AML) cell lines in vitro. High-resolution LC-MS was first used to analyze the composition of the aqueous extract. Cell proliferation assay, Annexin V staining, cell cycle analysis, dual Annexin V/PI staining, cell death quantification by ELISA, ROS level detection and Western Blotting were then performed to elucidate the therapeutic effects of AAL extract. The results obtained revealed a potent antioxidant activity of AAL extract. Moreover, the extract exhibited dose- and time-dependent antiproliferative effects on AML cell lines by decreasing cell viability with an IC50 of 5.03% (v/v) at 24 h of treatment of KG-1 cells. This decrease in viability was accompanied with a significant increase in apoptotic cell death with cell cycle arrest and flipping of the phosphatidylserine from the inner to the outer leaflet of the cell membrane. The respective overexpression and downregulation of proapoptotic proteins like cleaved caspase-8, cleaved PARP-1 and Bax and antiapoptotic proteins like Bcl-2 further validated the apoptotic pathway induced by AAL on AML cells. Finally, LC-MS revealed the presence of several compounds like fatty acids, terpenes, phenolics, cinnamic acids and flavonoids that could contribute to the antioxidant and anti-cancer effects of this herbal infusion. In addition to the generally known nutritional effects of the Annona cherimola fruit and leaves, the presented data validates the antioxidant and anti-cancerous effects of the leaf infusion on AML cell lines, proposing its potential therapeutic use against acute myeloid leukemia with future in vivo and clinical trials.

ACS Style

Tony Haykal; Maria Younes; Marianne El Khoury; Carl Ammoury; Stephanie Tannous; Mohammad H. Hodroj; Rita Sarkis; Natalia Gasilova; Laure Menin; Sandra Rizk. The pro-apoptotic properties of a phytonutrient rich infusion of A. cherimola leaf extract on AML cells. Biomedicine & Pharmacotherapy 2021, 140, 111592 .

AMA Style

Tony Haykal, Maria Younes, Marianne El Khoury, Carl Ammoury, Stephanie Tannous, Mohammad H. Hodroj, Rita Sarkis, Natalia Gasilova, Laure Menin, Sandra Rizk. The pro-apoptotic properties of a phytonutrient rich infusion of A. cherimola leaf extract on AML cells. Biomedicine & Pharmacotherapy. 2021; 140 ():111592.

Chicago/Turabian Style

Tony Haykal; Maria Younes; Marianne El Khoury; Carl Ammoury; Stephanie Tannous; Mohammad H. Hodroj; Rita Sarkis; Natalia Gasilova; Laure Menin; Sandra Rizk. 2021. "The pro-apoptotic properties of a phytonutrient rich infusion of A. cherimola leaf extract on AML cells." Biomedicine & Pharmacotherapy 140, no. : 111592.

Journal article
Published: 13 November 2020 in BMC Complementary Medicine and Therapies
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Background Herbal medicines have been a major target for numerous studies through the past years as an alternative treatment for cancer, mainly due to their minimal effects on normal healthy cells. Annona cherimola, popularly known as Cherimoya, is an edible natural fruit rich in phytochemical components and known to possess various biological activities. Previous studies have reported the anti-cancerous effect of A. cherimola ethanolic leaf extract (AELE) on leukemia. This study aims at studying the potential anti-cancer activity of this extract in vitro in two different breast cancer cell lines, namely MDA-MB-231 and MCF-7, in addition to investigating its toxicity on normal mesenchymal stem cells. Methods The anti-proliferative effect of AELE was evaluated via cell viability assay. Propidium iodide staining, Cell Death Detection ELISA and flow cytometry analysis of Annexin V binding were used to assess cell cycle progression, DNA fragmentation and apoptosis induction, respectively. Protein expression was determined via Western Blot analysis to decipher the underlying apoptotic molecular mechanism induced upon AELE treatment. Results The anti-proliferative effect of the extract was found to be selective on the triple-negative breast cancer cell line (MDA-MB-231) in a time- and dose-dependent manner with an IC50 of 390.2 μg/mL at 48 h, with no cytotoxic effects on normal murine mesenchymal stem cells. The pro-apoptotic effect was confirmed by the increase in cellular and DNA fragmentation, flipping of the phosphatidylserine moiety to the outer leaflet, and the increase in Annexin V binding. The underlying molecular mechanism revealed the involvement of the mitochondrial pathway, as shown by alterations in mitochondrial permeability and the upregulation of cytochrome c expression. Conclusion All the data presented in our study suggest that AELE exhibits a selective anti-proliferative and pro-apoptotic effect on the chemo-resistant MDA-MB-231 breast cancer cells, providing evidence for the anti-tumor effects of A. cherimola.

ACS Style

Maria Younes; Carl Ammoury; Tony Haykal; Leah Nasr; Rita Sarkis; Sandra Rizk. The selective anti-proliferative and pro-apoptotic effect of A. cherimola on MDA-MB-231 breast cancer cell line. BMC Complementary Medicine and Therapies 2020, 20, 1 -10.

AMA Style

Maria Younes, Carl Ammoury, Tony Haykal, Leah Nasr, Rita Sarkis, Sandra Rizk. The selective anti-proliferative and pro-apoptotic effect of A. cherimola on MDA-MB-231 breast cancer cell line. BMC Complementary Medicine and Therapies. 2020; 20 (1):1-10.

Chicago/Turabian Style

Maria Younes; Carl Ammoury; Tony Haykal; Leah Nasr; Rita Sarkis; Sandra Rizk. 2020. "The selective anti-proliferative and pro-apoptotic effect of A. cherimola on MDA-MB-231 breast cancer cell line." BMC Complementary Medicine and Therapies 20, no. 1: 1-10.

Journal article
Published: 13 February 2020 in Cancers
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Malva pseudolavatera Webb & Berthel. is a plant from the Malvaceae family that has long been included in the human diet due to its various curative effects. Many plant leaf extracts from the various species of Malva genus have been reported to possess anti-cancer properties, however, studies on M. pseudolavatera Webb & Berthel. leaves have documented anti-inflammatory and anti-oxidant effects with no emphasis on their possible anti-cancer potential. The present study explores the anti-cancer properties of Malva pseudolavatera Webb & Berthel. leaf extract on acute myeloid leukemia (AML) cell lines in vitro and deciphers the underlying molecular mechanism. Treatment of AML cell lines with M. pseudolavatera methanolic leaf extract showed a dose- and time-dependent inhibition of proliferation and a dose-dependent increase in apoptotic hallmarks such as an increase in phosphatidylserine on the outer membrane leaflet and membrane leakage in addition to DNA fragmentation. The pro-apoptotic effect was induced by reactive oxygen species (ROS) as well as an upregulation of cleaved poly(ADP-ribose) polymerase (PARP), increase in Bax/Bcl-2 ratio, andrelease of cytochrome-c from the mitochondria. Major compounds of the extract included methyl linolenate, phytol, γ-sitosterol, and stigmasterol as revealed by gas chromatography coupled with mass spectrometry, and amino acids, amino acid derivatives, tiliroside, 13-hydroxyperoxyoctadecadienoic, and quercitrin as detected by liquid chromatography coupled to mass spectrometry.

ACS Style

Marianne El Khoury; Tony Haykal; Mohammad H. Hodroj; Sonia Abou Najem; Rita Sarkis; Robin I. Taleb; Sandra Rizk. Malva pseudolavatera Leaf Extract Promotes ROS Induction Leading to Apoptosis in Acute Myeloid Leukemia Cells In Vitro. Cancers 2020, 12, 435 .

AMA Style

Marianne El Khoury, Tony Haykal, Mohammad H. Hodroj, Sonia Abou Najem, Rita Sarkis, Robin I. Taleb, Sandra Rizk. Malva pseudolavatera Leaf Extract Promotes ROS Induction Leading to Apoptosis in Acute Myeloid Leukemia Cells In Vitro. Cancers. 2020; 12 (2):435.

Chicago/Turabian Style

Marianne El Khoury; Tony Haykal; Mohammad H. Hodroj; Sonia Abou Najem; Rita Sarkis; Robin I. Taleb; Sandra Rizk. 2020. "Malva pseudolavatera Leaf Extract Promotes ROS Induction Leading to Apoptosis in Acute Myeloid Leukemia Cells In Vitro." Cancers 12, no. 2: 435.

Journal article
Published: 12 December 2019 in BMC Complementary and Alternative Medicine
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Background The edible fruit Annona cherimola has previously shown many nutritional and medicinal properties. The current study evaluates the anti-cancer and anti-proliferative properties of Annona cherimola ethanolic leaf extract (AELE) on Acute Myeloid Leukemia (AML) cell lines cultured in vitro (Monomac-1 and KG-1). Methods The anti-proliferative effect of A. cherimola ethanolic leaf extract was evaluated via cell viability assay. Its pro-apoptotic effect was assessed through Cell Death ELISA and dual Annexin V/PI staining. To further investigate the molecular mechanism by which the extract promoted apoptosis and inhibited the proliferation of the AML cells used, apoptotic protein expression was determined through western blots. Extract composition was elucidated by Gas Chromatography-Mass Spectrometry (GC-MS). Results Our results showed that the treatment with A. cherimola ethanolic leaf extract exhibited an inhibitory effect on the proliferation of both cancer cell lines used in a dose- and time-dependent manner, with no toxic effects on normal mononuclear cells (MNCs) isolated from human bone marrow. This effect was mediated by DNA fragmentation and apoptosis, as revealed by Cell Death ELISA and dual Annexin V/PI staining. Western blot analysis revealed a Bax/Bcl2 dependent mechanism of apoptosis, as well as PARP cleavage, confirming the apoptotic results observed previously. These effects may be attributed to the presence of terpenes which constitute a large component of the leafy extract, as revealed via GC-MS. Conclusion All the data presented in our study show that the terpene-rich A. cherimola ethanolic leaf extract exhibits an anti-proliferative and pro-apoptotic effect on the AML cell lines used.

ACS Style

Carl Ammoury; Maria Younes; Marianne El Khoury; Mohammad H. Hodroj; Tony Haykal; Peter Nasr; Marilyne Sily; Robin I. Taleb; Rita Sarkis; Rana Khalife; Sandra Rizk. The pro-apoptotic effect of a Terpene-rich Annona cherimola leaf extract on leukemic cell lines. BMC Complementary and Alternative Medicine 2019, 19, 1 -10.

AMA Style

Carl Ammoury, Maria Younes, Marianne El Khoury, Mohammad H. Hodroj, Tony Haykal, Peter Nasr, Marilyne Sily, Robin I. Taleb, Rita Sarkis, Rana Khalife, Sandra Rizk. The pro-apoptotic effect of a Terpene-rich Annona cherimola leaf extract on leukemic cell lines. BMC Complementary and Alternative Medicine. 2019; 19 (1):1-10.

Chicago/Turabian Style

Carl Ammoury; Maria Younes; Marianne El Khoury; Mohammad H. Hodroj; Tony Haykal; Peter Nasr; Marilyne Sily; Robin I. Taleb; Rita Sarkis; Rana Khalife; Sandra Rizk. 2019. "The pro-apoptotic effect of a Terpene-rich Annona cherimola leaf extract on leukemic cell lines." BMC Complementary and Alternative Medicine 19, no. 1: 1-10.

Journal article
Published: 30 August 2019 in Toxins
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Annona cherimola Mill is a large green fruit with black seeds widely known to possess toxic properties due to the presence of Annonaceous acetogenins. The present study investigates the anti-cancer properties of an Annona cherimola Mill ethanolic seed extract on Acute Myeloid Leukemia (AML) cell lines in vitro and elucidates the underlying cellular mechanism. The anti-proliferative effects of the extract on various AML cell lines and normal mesenchymal cells (MSCs) were assessed using WST-1 viability reagent. The pro-apoptotic effect of the extract was evaluated using Annexin V/PI staining and Cell Death ELISA. The underlying mechanism was deciphered by analyzing the expression of various proteins using western blots. Treatment with an A. cherimola seed ethanolic extract promotes a dose- and time-dependent inhibition of the proliferation of various AML cell lines, but not MSCs. Positive Annexin V staining, as well as DNA fragmentation, confirm an increase in apoptotic cell death by upregulating the expression of pro-apoptotic proteins which control both intrinsic and extrinsic pathways of apoptosis. GC/MS analysis revealed the presence of phytosterols, in addition to other bioactive compounds. In conclusion, Annona cherimola Mill seed extract, previously known to possess a potent toxic activity, induces apoptosis in AML cell lines by the activation of both the extrinsic and the intrinsic pathways.

ACS Style

Tony Haykal; Peter Nasr; Mohammad H. Hodroj; Robin I. Taleb; Rita Sarkis; Marvy Nadine El. Moujabber; Sandra Rizk. Annona cherimola Seed Extract Activates Extrinsic and Intrinsic Apoptotic Pathways in Leukemic Cells. Toxins 2019, 11, 506 .

AMA Style

Tony Haykal, Peter Nasr, Mohammad H. Hodroj, Robin I. Taleb, Rita Sarkis, Marvy Nadine El. Moujabber, Sandra Rizk. Annona cherimola Seed Extract Activates Extrinsic and Intrinsic Apoptotic Pathways in Leukemic Cells. Toxins. 2019; 11 (9):506.

Chicago/Turabian Style

Tony Haykal; Peter Nasr; Mohammad H. Hodroj; Robin I. Taleb; Rita Sarkis; Marvy Nadine El. Moujabber; Sandra Rizk. 2019. "Annona cherimola Seed Extract Activates Extrinsic and Intrinsic Apoptotic Pathways in Leukemic Cells." Toxins 11, no. 9: 506.