Antiproliferation Activity and Apoptotic Mechanism of Soursop (Annona muricata L.) Leaves Extract and Fractions on MCF7 Breast Cancer Cells

Abstract

Introduction: Breast cancer is the second most common cancer in women globally, and the incidence rate has increased annually. Traditional medicine is frequently used as a cancer treatment, and soursop or Annona muricata L (A. muricata) is a traditional medicinal plant that has been widely used as an anticancer treatment and requires more thorough study. Methods: In this research, we prepared ethanol extract and three solvents, ie, ethyl acetate, n-hexane and water fractions of A. muricata leaves and assessed their antiproliferation and cytotoxic activity on MCF7 breast cancer cells compared with that on CV1 normal kidney cells; observation of cell morphology by stained with mixture of propidium iodide and 4ʹ,6-diamidino-2-phenylindole indicated that this treatment induced an ongoing process of apoptotic cell death in MCF7 cells. To clarify the cell death mechanism via apoptosis, we assessed the mRNA expression in the caspase cascade of caspase-9, caspase-3, and PARP-1, and anti-apoptotic, Bcl-2 which mediated cytotoxic activity of extracts and ethyl acetate fractions of A. muricata leaves against MCF7 cells. Results: The ethanol extract, ethyl acetate, n-hexane, and water fractions of A. muricata leaves had IC 50 values of 5.3, 2.86, 3.08, and 48.31 µg/mL, respectively, in MCF7 cells but had no activity in CV1 cells. The high cytotoxic activity of A. muricata leaves was reflected by changes in the morphology of cancer cells that appeared after 6 h exposure to A. muricata leaf extract and ethyl acetate fraction; the membrane and nucleus of cells undergoing apoptosis were characterized by the rupture and loss of membranes and nuclei. The mechanism that mediates this cytotoxic activity in MCF7 cells was mediated through a decrease in the expression of Bcl-2 mRNA and an increase in caspase-9 and caspase-3 mRNA expression. Conclusion: Therefore, the leaves of the medicinal plant A. muricata contained compounds that on extraction exerted a highly effective activity as an anticancer treatment for breast cancer via induced apoptotic cell death.

Pharmacological Activities of Soursop (Annona muricata Lin.)

“…The leaf extract induced apoptosis by enhancing the expression of caspase-3 in the colorectal cancer cell line COLO-205 [19] and the breast cancer cell line MDA-MB-231 [20]. Another study showed that the ethanol extract and ethyl acetate fractions of A. muricata leaves were active against MCF7 cells via an apoptosis mechanism mediated by decreased Bcl-2 expression and increased caspase-3 and caspase-9 expression [23]. Additionally, the presence of annonaceous acetogenins along with flavonoids in A. muricata leaves inhibited the proliferation of the human prostate cancer cell line PC-3 [21].…”

“…[22] Apoptosis mechanism mediated by a decrease in Bcl-2 expression and an increase in caspase-3 and caspase-9 expression in MCF7 breast cancer cells. [23] Seeds Increases caspase-3 cleavage and DNA fragmentation in endometrial cancer cells. [24] Antiulcer Leaf…”

A Review on Annona muricata and Its Anticancer Activity

“…Acetogenin’s mechanism for inducing the apoptotic process involves blocking ATP production by inhibiting the NADH: ubiquinone oxidoreductase (Complex I) in the electron transport system in the mitochondria of cancer cells. This results in the cancer cells losing their supply of energy and becoming weaker, ultimately leading to cell death [ 61 , 62 ]. Including the methyl group on the γ-lactone moiety makes natural acetogenins potent mitochondrial complex I inhibitors [ 63 ].…”

“…Exposure of cancer cells to A. muricata leaf extract and ethyl acetate fraction resulted in morphological alterations indicative of apoptosis, a process characterized by the rupture and loss of the membrane and nucleus of cells. Reduced Bcl-2 and PARP-1 and increased caspase-9 and caspase-3 expression are responsible for the cytotoxic activity observed in MCF7 cells [ 62 ]. Treatment of Annonacin (0.5–1 µM) induced MCF breast cancer cell line death at 48 h and treatment of Annonacin (0.1 µM) in xenografts tumor decreases the expression of ER, cyclin D1, and Bcl2 ( Figure 5 ) [ 135 ].…”

Nutritional and Therapeutic Potential of Soursop

“…Proteolytic enzymes are activated by cells which should kill during the apoptosis process. Apoptosis plays a role in suppressing the progression of liver disease, either through the intrinsic or extrinsic pathway, as well as disorders of the mitochondria [53,54]. is is a crucial physiological function that is necessary for tissue homeostatic processes and normal cell maintenance [53].…”