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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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Abstract

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.

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Abbreviations

BCA:

bicinchoninic acid

CF:

chloroform fraction

DMEM:

Dulbecco’s modified Eagle’s medium

DMSO:

dimethyl sulfoxide

FBS:

fetal bovine serum

FID:

flame ionization detector

GC:

gas chromatography

HF:

n-hexane fraction

IgG:

immunoglobulin G

MS:

mass spectrometry

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

PI:

propidium iodide

PVDF:

polyvinylidene fluoride

RT:

room temperature

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Authors and Affiliations

  1. Faculty of Biotechnology, Cheju National University, 690-756, Jeju, Republic of Korea

    Jeong Yong Moon, Hana Kim & Somi Kim Cho

  2. Department of Medicine, Cheju National University, 690-756, Jeju, Republic of Korea

    Moonjae Cho & Weon Young Chang

  3. Food Safety Research Institute, Nong Shim Co., LTD, 156-709, Seoul, Republic of Korea

    Cheong-Tae Kim

  4. The Research Institute for Subtropical Agriculture and Biotechnology, Cheju National University, 690-756, Jeju, Republic of Korea

    Somi Kim Cho

  5. Subtropical Horticulture Research Institute, Cheju National University, 690-756, Jeju, Republic of Korea

    Somi Kim Cho

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  1. Jeong Yong Moon
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  2. Hana Kim
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  3. Moonjae Cho
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  4. Weon Young Chang
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  5. Cheong-Tae Kim
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  6. Somi Kim Cho
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Correspondence to Somi Kim Cho.

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Yong Moon, J., Kim, H., Cho, M. et al. Induction of Apoptosis in SNU-16 Human Gastric Cancer Cells by the Chloroform Fraction of an Extract of Dangyuja (Citrus grandis Leaves. J. Korean Soc. Appl. Biol. Chem. 52, 168–175 (2009). https://doi.org/10.3839/jksabc.2009.031

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  • DOI: https://doi.org/10.3839/jksabc.2009.031

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