| Objective To study the mechanism of the expression of MMP-2 gene and the effect on the activity of MMP-2 induced by FN on Hela cell. To discuss whether the Ganoderma polysaccharide can inhibit metastasis of tumor through inhibiting the activity of MMP-2. Methods Hela cell was cultured in RPIM1640 without calf serum and FN worked as extracellular matrix, then Hela cell was induced by different concentration of FN and at the same concentration of FN in different periods. Detect the adhesive rate of Hela cell by MTT; count the spread rate and observe the structure of Hela cell under the inverted microscope; measure the activity of MMP-2 by gelatin zymography and detect the expression of MMP-2 gene by RT-PCR. Antisense ODN was used for blocking the focal adhesion kinase which was the key enzyme during the signal transduction mediated by FN-integrin. Observe the variety of activity of MMP-2. Ganoderma polysaccharide was also used in this experiment, then observe the effect of activity of MMP-2. Results When there was no FN, no adhesion and spread of Hela cell, no expression of MMP-2 gene and no activity of MMP-2; when the concentration of FN reached 5~10ug/ml, the adhesive rate and the spread rate were the highest. When the FN was used for 4 hours with the concentration of lOug/ml, MMP-2 gene began to express; and for 12 hours with the same concentration, the activity of MMP-2 was the highest. The activity of MMP-2 was dramatically decreased when pp125FAK antisense ODN was put into the culture medium. Conclusion Hela cell can be induced by FN to adhere, which causes a series of signal transductions in cells, promotes the expression of MMP-2 gene and enhances the activity of MMP-2. Perhaps this reaction happens through Grb2/Ras-MAPKs, whose keyingredient was FAK. Ganoderma polysaccharide can inhibit the activity of MMP-2, thereby inhibit metastasis of tumor. But the mechanism will be further studied in future. |
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