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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

  • Jeong Yong Moon
  • Hana Kim
  • Moonjae Cho
  • Weon Young Chang
  • Cheong-Tae Kim
  • Somi Kim ChoEmail author
Bioactive Materials

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.

Key words

apoptosis Bcl-2 family caspase-3 activity Dangyuja SNU-16 human gastric cancer cell 

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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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Jeong Yong Moon
    • 1
  • Hana Kim
    • 1
  • Moonjae Cho
    • 2
  • Weon Young Chang
    • 2
  • Cheong-Tae Kim
    • 3
  • Somi Kim Cho
    • 1
    • 4
    • 5
    Email author
  1. 1.Faculty of BiotechnologyCheju National UniversityJejuRepublic of Korea
  2. 2.Department of MedicineCheju National UniversityJejuRepublic of Korea
  3. 3.Food Safety Research InstituteNong Shim Co., LTDSeoulRepublic of Korea
  4. 4.The Research Institute for Subtropical Agriculture and BiotechnologyCheju National UniversityJejuRepublic of Korea
  5. 5.Subtropical Horticulture Research InstituteCheju National UniversityJejuRepublic of Korea

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