Abstract Title:

Phenolic Extraction ofLeaves Induces Caspase-Dependent and Caspase-Independent Apoptosis through the Generation of Reactive Oxygen Species and the Activation of Intrinsic Mitochondrial Pathway in Human Melanoma Cells.

Abstract Source:

Nutr Cancer. 2020 Jun 12:1-20. Epub 2020 Jun 12. PMID: 32530312

Abstract Author(s):

Bich Hang Do, Nghia Son Hoang, Thi Phuong Thao Nguyen, Nguyen Quynh Chi Ho, Thanh Long Le, Chinh Chung Doan

Article Affiliation:

Bich Hang Do


Lam. has long been used to treat many diseases, including diabetes, aging, inflammatory, and cancer. Many studies have revealed that the crude extract ofLam. leaves possesses anticancer property. Therefore, in this study, the extract ofleaves was fractionated using different solvents to figure out the most effective fraction for anti-proliferative effect on melanoma cells. Methanol extract (MO-ME), hexane fraction (MO-HE), chloroform fraction (MO-CH), ethyl acetate fraction (MO-EA), and water-soluble fraction (MO-WA) ofleaves were prepared. Total phenolic and flavonoid contents were determined. The anti-proliferative activity on melanoma cells and normal cells was investigated using WST-1 assay. The apoptotic activity was assessed by testing DNA condensation, DNA fragmentation, and phosphatidylserine (PS) externalization. The expression of apoptosis-related genes, the mitochondrial depolarization, and reactive oxygen species (ROS) were then examined to clarify the underlying molecular mechanisms. In this regard, MO-ME, MO-EA, and MO-CH inhibited the proliferation of both A375 human melanoma cells and A2058 human melanoma cells, but had little effect on WS1 normal human skin fibroblasts and primary normal human dermal fibroblasts (NHDF). Among fractions, the phenolic-rich MO-EA markedly inhibited the growth of A375 cells in a dose- and time-dependent manner. The anti-proliferation was supposed to be mediated via apoptosis, which was demonstrated by the significant increase of condensed chromatin, DNA fragmentation, and PS externalization. The apoptosis was stimulated by enhanced ROS production and reduction of mitochondrial membrane potential. MO-EA activated Bax while reducing Bcl-2 expression, leading to an increase in Bax/Bcl-2 ratio. The mechanisms of cell death involved in activation of Caspase-3/7 and Caspase-9 (Caspase-dependent pathway), activation, and translocation of apoptosis-inducing factor (AIF) into the nucleus (Caspase-independent pathway). Our study indicated that the phenolic-rich fraction exerted significant anticancer effects on melanoma cellswhich involved in Caspase-dependent and Caspase-independent apoptosis pathways mediated by mitochondrial ROS. These results provided a fundament for the using of phenolic-rich fraction ofleaves to treat skin cancer effectively.

Study Type : In Vitro Study
Additional Links
Pharmacological Actions : Apoptotic : CK(6986) : AC(5304)

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