Matrix metalloproteinases are family of zinc dependent endoproteinases degrade the extracellular matrix, have been identified as poor prognosis markers and therapeutic targets for breast cancer patients. Effect of apigenin was observed on mRNA expression on different MMPs in MDA MB- 231 breast cancer cells including its effect on cancer cell migration and invasion.
MDA MB- 231 cells were cultured with different concentrations (0, 10, 20 and 40 μM) of apigenin for 48 hrs. mRNA expressions of MMP-1, MMP-3, MMP-7, MMP-9 amd MMP- 11 were analyzed by real time PCR. For cell migration wound healing assay was done at 0, 6, 24, and 48 h periods. To asses the cell invasion MDA MB-231 cancer cells were seeded on collagen based matrigel chamber and treated with 10, 20 and 40μM apigenin, whereas untreated cells considered as control. After 48hrs cells invaded through matrigel quantified by colorimetric method.
Our results showed that MMP-1 and MMP-11 were significantly downregulated upto 0.36 folds and 0.55 folds respectively at 40 μM apigenin treatment in MDA MB-231 cells in comparision to cells without drug. In comparision to untreated cancer cells the MMP-3 gene expressions were 0.46, 0.27 and 0.12 fold in 10μM, 20μM and 40μM apigenin treated MDA MB-231 cells. The apigenin significantly decreased MMP-7 gene expression level upto 0.34 fold and 0.05 fold at 20 μM and 40 μM concentrations in MDA MB-231 cells. Apigenin significantly downregulated MMP-9 mRNA expression level in dose dependent manner in MDA MB-231 cancer cells as compared to untreated cells. Wound healing assay results exhibited that apigenin significantly reduce the migration of cancer cells. In cells treated with 20μM and 40μM apigenin 16.64% and 25.81% of the wound remains unresolved after 48 hrs. The cell invasion assay showed that apigenin significantly inhibited the migration of cells, 40 μM of apigenin showed only 73.52% invasion, as compared to the untreated cells.
In conclusion, apigenin reduced the mRNA expressions of MMP-1, MMP-3, MMP-7, MMP-9 and MMP-11 in MDA-MB-231 cells. Apigenin also inhibited the invasion and cell migration. Therefore, we suggest that apigenin may be used as a candidate drug for the inhibition of metastasis of human breast cancer.
This is the part of my Ph. D. degree, done at Animal Biochemistry Division, ICAR- Indian Veterinary Reasech Institute, Izatnagar, Bareilly, U.P., India
Dr. Shweta Rajoriya.
ICAR-Indian Veterinary Research Institute, Bareilly, U.P., India (Indtitutional Funding).
The author has declared no conflicts of interest.