Nutr Res Pract.  2011 Oct;5(5):375-380.

Inorganic sulfur reduces the motility and invasion of MDA-MB-231 human breast cancer cells

Affiliations
  • 1Department of Food Science and Nutrition, Dankook University, 126 Jukjeon-dong, Suji-gu, Yongin-si, Gyunggi 448-701, Korea. wkkim@dankook.ac.kr
  • 2Department of Food and Nutrition, Beawha Women's University, Seoul 110-735, Korea.

Abstract

This study investigated the effects of inorganic sulfur on metastasis in MDA-MB-231 human breast cancer cells. MDA-MB-231 cells were cultured in the absence or presence of various concentrations (12.5, 25, or 50 micromol/L) of inorganic sulfur. Cell motility, invasion, and the activity and mRNA expression of matrix metalloproteases (MMPs) were examined. Numbers of viable MDA-MB-231 cells did not differ by inorganic sulfur treatment from 0 to 50 micromol/L within 48 h. Inorganic sulfur significantly decreased cell motility and invasion in the MDA-MB-231 cells in a dose-dependent manner (P < 0.05), as determined using a Boyden chamber assay and a Matrigel chamber. The activities of MMP-2 and MMP-9 were significantly reduced by inorganic sulfur in a dose-dependent manner (P < 0.05). The inorganic sulfur also significantly inhibited MMP-2 and MMP-9 expression in the cells (P < 0.05). These data suggest that inorganic sulfur can suppress cancer cell motility and invasion by inhibiting MMP-2 and MMP-9 activity and gene expression in MDA-MB-231 cells.

Keyword

Inorganic sulfur; invasion; motility; matrix metalloprotease; MDA-MB-231 cells

MeSH Terms

Breast
Breast Neoplasms
Cell Movement
Collagen
Drug Combinations
Gene Expression
Humans
Laminin
Metalloproteases
Neoplasm Metastasis
Proteoglycans
RNA, Messenger
Sulfur
Collagen
Drug Combinations
Laminin
Metalloproteases
Proteoglycans
RNA, Messenger
Sulfur

Figure

  • Fig. 1 Effect of inorganic sulfur on cell proliferation in MDA-MB-231 cells. MDA-MB-231 cells were plated at a density of 2.5 × 104 cells/mL in 24 well plates with DMEM/F12 supplemented with 10% FBS for 48 h. The monolayers were serum-starved with DMEM/F12 supplemented with 5 µg/mL transferrin, 5 ng/mL selenium, and 1 mg/mL bovine serum albumin for 24 h. After serum starvation, the monolayer were incubated in serum free medium with 0, 12.5, 25, or 50 µmol/L inorganic sulfur for 24 h. Viable cell numbers were estimated by the MTT assay. Each bar represents the mean ± SE from 3 independent experiments. Significant differences (P < 0.05) among groups are indicated by different letters above each bar.

  • Fig. 2 Effect of inorganic sulfur on cell motility in MDA-MB-231 cells. Cells were cultured in the presence of various concentrations of inorganic sulfur (0, 12.5, 25, or 50 µmol/L) for 8 h with a Boyden chamber. A) Microphotography of cells treated with inorganic sulfur in cell migration assay. B) Quantitative analysis of cell motility assay. The number of migrated cells was expressed as a percentage of the control (without inorganic sulfur). Each bar represents the mean ± SE from 3 independent experiments. Significant differences (P < 0.05) among groups are indicated by different letters above each bar.

  • Fig. 3 Effects of inorganic sulfur on invasion in MDA-MB-231 cells. Cells were cultured in the presence of various concentrations of inorganic sulfur (0, 12.5, 25, 50 µmol/L) for 12 h in an invasion chamber. A) Microphotography of cells treated with inorganic sulfur in cell migration assay. B) Quantitative analysis of cell motility assay. The number of migrated cells was expressed as a percentage of the control (without inorganic sulfur). Each bar represents the mean ± SE from 3 independent experiments. Significant differences (P < 0.05) among groups are indicated by different letters above each bar.

  • Fig. 4 Effects of inorganic sulfur on MMP-2 and MMP-9 activity in MDA-MB-231 cells. Cells were plated in 6 well plates at a density 1 × 103 cell/mL in DMEM/F12 supplemented with 10% FBS. After 48 h of incubation, the monolayers were incubated in the presence of various concentrations of inorganic sulfur (0, 12.5, 25, 50 µmol/L) for 24 h. The medium was collected and concentrated for zymography. A-a) Photographs of MMP-2, A-b) Quantitative analysis of the bands. B-a) Photographs of MMP-9 bands, B-b) Quantitative analysis of the bands. The enzyme activity was expressed as a percentage of the control (without inorganic sulfur). Each bar represents the mean ± SE of 3 independent experiments. Significant differences (P < 0.05) among groups are indicated by different letters above each bar.

  • Fig. 5 Effect of inorganic sulfur on MMP-2 and MMP-9 mRNA expression in MDA-MB-231 cells. Cells were cultured in the presence of various concentrations of inorganic sulfur (0, 12.5, 25, or 50 µmol/L) for 24 h. Total RNA was isolated and RT-PCR was performed. Photographs of the ethidium bromide-stained gels, which are representative of 3 independent experiments, are shown. A-a) Photographs of the MMP-2 bands, A-b) Quantitative analysis of the bands. B-a) Photographs of the MMP-9 bands, B-b) Quantitative analysis of the bands. The mRNA expression of the enzymes was expressed as a percentage of the control (without inorganic sulfur). Each bar represents the mean ± SE, and significant differences (P < 0.05) among groups are indicated by different letters above each bar.


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