Induction of Apoptosis in SNU-16 Human Gastric Cancer Cells by the Chloroform Fraction of an Extract of Dangyuja (Citrus grandis Leaves
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The anticancer effect of Dangyuja (Citrus grandis Osbeck) leaf extract was investigated using SNU-16 human gastric cancer cells. Maximum cytotoxicity was observed from the chloroform fraction (CF) of the extract. Cell death was dose-dependent (IC50=ca. 92.15 μg/mL), and was characterized by apoptotic body formation and DNA fragmentation. Flowcytometric analysis showed that treatment of CF resulted in a marked accumulation of cells in the sub-G1 phase. The induction of apoptosis was confirmed by caspase-3 activity assays and immunoblotting using antibodies against Bid, Bcl-2, Bax, poly (ADP-ribose) polymerase, caspase-9, caspase-8, caspase-7, and caspase-3. Analyses of the CF by gas chromatography (GC) and GC-mass spectrometry tentatively identified 21 compounds, including linoleic acid, linolenic acid, palmitic acid, α-amyrin, γ-sitosterol, dihydrolanosterol, methyl palmitate, and nobiletin. These results provided the first evidence that the CF of a Dangyuja leaf extract induces apoptosis in SNU-16 cells. Our findings may lead to new strategies for the treatment of human gastric cancer.
Key wordsapoptosis Bcl-2 family caspase-3 activity Dangyuja SNU-16 human gastric cancer cell
Dulbecco’s modified Eagle’s medium
fetal bovine serum
flame ionization detector
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