Immune Netw.  2011 Aug;11(4):210-215. 10.4110/in.2011.11.4.210.

The Anti-tumor Activity of Vitamin C via the Increase of Fas (CD95) and MHC I Expression on Human Stomach Cancer Cell Line, SNU1

• Yu Y
• Bae S
• Kim H
• Kim Y
• Chu NB
• Chu NK
• Kang JS
• Lee WJ

• KMID: 1449812
• DOI: http://doi.org/10.4110/in.2011.11.4.210

Abstract

BACKGROUND
It is already known that high concentration of vitamin C induces apoptosis on tumor cells. However, there is no report regarding the function of vitamin C on the modulation of immune susceptibility of cancer. Therefore, we investigated whether vitamin C can modulate immune susceptibility of tumor cells, especially on the induction of Fas-mediated apoptosis.
METHODS
First, the optimal concentration of vitamin C, which cannot induce damages on tumor cells for 36 hrs. We found that 2 mM of vitamin C did not show harmful effect. In addition, the optimal concentration of agonistic anti-Fas Abs for 18 hrs was examined.
RESULTS
As a result, 400 ng/ml of agonistic anti-Fas Abs did not induce apoptosis on tumor cells. Next, we tried to find the effect of 2 mM of vitamin C on the modulation of the susceptibility to agonistic anti-Fas Abs. When tumor cells were cultured with 400 ng/ml of agonistic anti-Fas Abs for 18 hrs, after pre-treatment with 2 mM of vitamin C for 24 hrs, viability of cells was decreased. Interestingly, we found that the expression of Fas (CD95) and MHC class I was increased by the treatment of vitamin C.
CONCLUSION
Taken together, vitamin C increases the susceptibility of tumor cells to anti-Fas Abs and the expression of Fas (CD95) and MHC class I on tumor cells.

Keyword

Vitamin C; Antitumor activity; Stomach cancer; Fas

MeSH Terms

Apoptosis
Ascorbic Acid
Cell Line
Humans
Stomach
Stomach Neoplasms
Vitamins
Ascorbic Acid
Vitamins

Figure

• 

  Figure 1 Investigation of the optimal doses of vitamin C. Human stomach cancer cell line, SNU1(1×106 cells) was in the presence or absence of vitamin C (1 and 2 mM) for 36 hrs. The cells were collected and washed twice with cold PBS. After the addition of 5µl of Annexin V-FITC, cells were incubated at room temperature for 15 min in the dark with gentle vortexing. One microliter of propidium of iodide was added, prior to analyze with FACSCaliber. Results are representative of three independent experiments.

• 

  Figure 2 Increase of Fas and MHC I expression on SNU1 by the treatment of vitamin C. The changing of Fas expression (A) and MHC expression (B) on human stomach cancer cell line, SNU1 by the treatment of vitamin C was assessed by flow cytometry. SNU1 (1×106 cells) was cultured in the presence or absence of vitamin C (2 mM) for 3, 6, 12, and 24 hrs. And then Cells were washed twice with phosphate buffered saline (PBS) and then stained with 1µg of PE-conjugated anti-human Fas mAbs or PE-conjugated anti-human MHC I mAbs for 30 min on ice. After washing twice with PBS, the cells were analyzed by FACSCaliber. Results are representative of three independent experiments.

• 

  Figure 3 Synergistic effect of vitamin C with anti-Fas mAb on the changing of cell viability in SNU1. (A) Optimal dose titration of anti-Fas mAb; Human stomach cancer cell line, SNU1 (1×106 cells) was cultured in the presence or absence of 200 and 400 ng/ml of anti-Fas mAbs (CH-11) for 18 hrs, and then cells were harvested and washed once with phosphate-buffered saline (PBS). Cell viability was examined by the trypan blue dye exclusion assay. Quadruplicate wells were run for each group. (B) Human stomach cancer cell line, SNU1 (1×106 cells) was cultured in the presence or absence of vitamin C (1 and 2 mM) for 24hrs, and then 200 and 400 ng/ml of anti-Fas mAbs (CH-11) were added. After further incubation for 18 hrs, cells were harvested and washed once with phosphate-buffered saline (PBS). Cell viability was examined by the trypan blue dye exclusion assay. Results are representative of three independent experiments. Quadruplicate wells were run for each group. Data represent means±SD.

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