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Inhibition of MMP-9-dependent cancer cell invasion
To investigate the role of TIMP-1 in the ovarian cancer cell invasion, a modified Boyden chamber assay was employed. The medium containing 10% FBS was used as an attractant in the lower chamber of Matrigel-coated transwell. As an MMP-9 inducer, TNF-α (10 ng/ml) was added in the upper chamber to facilitate the cancer cell invasion [25]. The number of the cancer Walrycin B invading through the Matrigel-coated membrane was increased by TNF-α as expected. However, the TNF-α-induced cancer cell invasion was greatly suppressed over 90% by the addition of TIMP-1 in the upper chamber (Fig. 3). Therefore, bark is possible to hypothesize that saxatilin suppresses the cancer cell invasion by TIMP-1-dependent regulation of MMP-9 activity.
Fig. 3.
MMP-9-dependent ovarian cancer cell invasion. MDAH 2774 human ovarian cancer cell invasion was stimulated with with TNF-α (10 ng/ml) on a transwell in the presence or absence of 50 nM saxatilin or TIMP-1 (10 ng/ml) and the invasion assay was preformed as described in Materials and methods.