J Korean Surg Soc.  2011 Aug;81(2):85-95. 10.4174/jkss.2011.81.2.85.

Induction of apoptosis with diallyl disulfide in AGS gastric cancer cell line

• Lee JE
• Lee RA
• Kim KH
• Lee JH

• KMID: 2096661
• DOI: http://doi.org/10.4174/jkss.2011.81.2.85

Abstract

PURPOSE
Diallyl disulfide (DADS) is a major organosulfur compound derived from garlic. It has been reported that DADS is able to inhibit the proliferation of several tumor cells. In this study, the effect of DADS was investigated in terms of the proliferation of AGS, gastric adenocarcinoma cell line at various concentrations.
METHODS
The viability of cultured cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. To detect the induction of apoptosis, Annexin V-FITC/propodium iodide (PI) staining assay was performed. Analysis of reactive oxygen species (ROS) and the distribution of cells in the cell cycle were measured by a flow cytometer. And using the Western blot analysis, the change of Fas, caspase-3, Bax, Bcl-2 activity was measured.
RESULTS
The percentage of live AGS cells was decreased to 23% of that in the control group after 400 microM DADS treatment for 48 hours. The Annexin V positive/PI negative (apoptosis portion) area increased from low concentration of DADS to high concentration. When comparing among the DADS treatment groups, the amount of ROS production increased in a dose dependent manner. The percentage of sub-diploid DNA content increased from 8.71% at 50 microM to 25.74% at 400 microM DADS treatment group. The expressions of Fas, caspase-3, Bax were increased and that of Bcl-2 was decreased in a dose dependent manner.
CONCLUSION
DADS decreases the viability of AGS cell lines and induces apoptosis in a dose-dependent manner. But the relationship of the anti-proliferative effect of DADS and related molecular changes were not clearly proportional to the concentration of DADS.

Keyword

Diallyl disulfide; Gastric cancer cell line; Apoptosis

MeSH Terms

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid
Adenocarcinoma
Allyl Compounds
Annexin A5
Apoptosis
Blotting, Western
Caspase 3
Cell Cycle
Cell Line
Cells, Cultured
Disulfides
DNA
Garlic
Reactive Oxygen Species
Stomach Neoplasms
4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid
Allyl Compounds
Annexin A5
Caspase 3
DNA
Disulfides
Reactive Oxygen Species

Figure

• 

  Fig. 1 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay of AGS cell viability after diallyl disulfide (DADS) treatment with 48 hours (P-value < 0.001).

• 

  Fig. 2 Diallyl disulfide induced apoptosis assessment with FACS analysis of AGS cells stained with Annexin V-FITC and propodium iodide. Apoptotic portion, lower right quadrant.

• 

  Fig. 3 The comparison of the results of proportion of apoptotic cells (Annexin V positive/propodium iodide negative cells) with flow cytometric analysis (P-value = 0.045).

• 

  Fig. 4 Reactive oxygen species production in AGS cells after treatment of diallyl disulfide with variable concentration.

• 

  Fig. 5 Comparison of the reactive oxygen species production between control group and diallyl disulfide (DADS) treatment group of variable concentration. MFI, mean fluorescence intensity.

• 

  Fig. 6 The percentage of sub-ciploid DNA-content of cell cycle induced by diallyl disulfide with flow cytometric analysis. M1, sub-diploid fraction and represents apoptotic cells; M2, G0-G1; M3, G2-M; M4, S phase of cells.

• 

  Fig. 7 Western blot in AGS cells after treatment of diallyl disulfide (DADS) with variable concentration.

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