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R.G.P.T. Jayasooriya*, M.G. Dilshara, Chang-Hee Kang, Hee-Ju Kim, Yung Hyun Choi, Gi-Young Kim
Laboratory of Immunobiology, Department of Marine Life Sciences, Jeju National University, Jeju 690-756,

Codium Fragile is a green macro algae belongs to family Codiaceae have been reported the anti tumor activity against various types of cancers. Aqueous extracts of Codium Fragile exhibited strong cytotoxic activities against CT26 mouse colon carcinoma cell line. The carotenoids which were isolated from Codium Fragile were shown cytotoxic effect on HL-60 human leukemia cells. We were interested in investigating the effects of Codium Fragile on MMP-9 gene expression in TNF-α-stimulated MDA-MB 231 human breast cancer cell line.

Codium Fragile (MECF) was extracted with distilled water at 100°C for 2 h. Human breast cancer cell line MDA-MB-231 was cultured in RPMI 1640. MTT assay was used to detect the effect of cell viability of MECF in MDA-MB-231 cells. RT-PCR was conducted to identify MECF regulated MMP-9 gene in breast cancer cells. We also detected the abilities of cell invasion and migration in breast cancer by performing invasion assay and gelatin zymography respectively. The expression of MMP-9 and NFκB related genes were analyzed by western blotting.

MECF had no effect on cell viability up to 100 μg/ml concentration in MDA-MB-231breast cancer cells regardless of the presence of TNF-α. The MDA-MB-231 cells which were stimulated with TNF-α showed a remarkable increase in cell invasion compaired with untreated control one while pretreatment with MECF down regulated the TNF-α-induced invasion of cells. Nevertheless, zymography, western blot and RT-PCR confirmed MECF down regulated TNF-α-induced MMP-9 upregulation. According to the electrophoretic morbidity shift assay, pretreatment with MECF significantly decreased TNF-α-induced DNA binding activity of NF-κB in MDAMB-231 cells. Moreover, treatment with MECF suppressed the TNF-α-induced expression of p65 and p50 in the cytosolic compartment. According to the luciferase assay, MECF remarkably decreased TNF-α-induced NF-κB luciferase activity.

MECF exhibit its anti-invasion effect by regulating Tumor necrosis Factor-α-Induced expression of Matrix Metalloproteinase-9 through the regulation of Nuclear Factor-κB pathway in human breast cancer cell line MDA-MB-231.

Keywords: Codium Fragile, Nuclear Factor-κB, Matrix Metalloproteinase-9, Tumor necrosis Factor-α

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