Role of Tissue Inhibitor of Metalloproteinases-2 (TIMP-2) in Regulation of Pro-Gelatinase A Activation Catalyzed by Membrane-Type Matrix Metalloproteinase-1 (MT1-MMP) in Human Cancer Cells

Abstract

To clarify the regulatory mechanism of pro-gelatinase A (proGelA) activation at a cellular level, expression of gelatinase A (GelA), three MT-MMPs, and TIMP-2 was examined with 11 human cancer cell lines cultured in the presence and absence of stimulants. MT1-MMP mRNA was expressed in 8 cell lines, while MT2-MMP and MT3-MMP mRNAs were expressed in fewer cell lines. The cells with high proGelA activation strongly expressed MT1-MMP mRNA but not MT2-MMP and MT3-MMP mRNAs, suggesting that MT1-MMP was responsible for the proGelA activation in the cancer cells. Treatments with concanavalin A (Con A) and a phorbor ester (TPA) enhanced the MT1-MMP expression, but only Con A stimulated the proGelA activation in many cell lines. In HT1080 fibrosarcoma cells, however, TPA also stimulated the activation. The level of TIMP-2 secreted into culture medium inversely correlated with proGelA activation. For example, 2 squamous cell carcinoma lines (HSC-3 and HSC-4) and 3 HT1080 clones, which efficiently activated proGelA, secreted little TIMP-2 into medium, whereas other cell lines and other HT1080 clones, which hardly activated proGelA, secreted TIMP-2 at high levels. When HSC-3 cells were incubated with TIMP-2 protein or transfected with TIMP-2 cDNA, the proGelA activation was strongly inhibited. These results indicated that extracellular TIMP-2 was an important negative regulator of proGelA activation. However, the level of extracellular TIMP-2 was not consistent with that of TIMP-2 mRNA in some cell lines. Other experimental results suggested that TIMP-2 might be rapidly metabolized after binding to MT1-MMP, and Con A treatment might stabilize the complex of TIMP-2 and MT1-MMP on cell membranes.

Prognostic Significance of Matrix Metalloproteinase 2 and Tissue Inhibitor of Metalloproteinase 2 Expression in Prostate Cancer

“…TIMP2 is reported to regulate matrix degradation, acting through a membrane type MMP (MT1-MMP; 55,56). MT1-MMP is a key enzyme in tumor angiogenesis and metastasis, hydrolyzes a variety of ECM components, and is a physiologic activator of pro-MMP2 (57).…”

Complex roles of tissue inhibitors of metalloproteinases in cancer

“…TIMP-2 can bind to proMMP-2 via C-terminal interaction, and to MT1-MMP by its N-terminal domain. This dual binding brings proMMP-2 close to cell surface, where it can be activated by neighboring TIMP-2-free MT1-MMP molecules (Shofuda et al, 1998;Butler et al, 1998). It has been reported that the entire propeptide domain of MT1-MMP is required for the TIMP-2 binding and subsequent proMMP-2 activation (Cao et al, 1998).…”

“…These contradictory facts promote one to investigate and rede®ne the net role of TIMPs on tumorigenesis. As summarized in Table 1 and Figure 1, in contrast to its well-established antitumor e ect, TIMP may also function in favor of tumor growth either in an MMP-independent or MMP-dependent manner, including (a) growth promotion and anti-apoptotic e ect (Hayakawa et al, 1992; Guedez et al, 1998;Li et al, 1999;Jiang et al, 2001); (b) inhibition of angiostatin and endostatin-converting MMPs or upregulation of VEGF (Yoshiji et al, 1998), and therefore stimulation of angiogenesis; and (c) involvement of activation of pro-MMP-2 (Shofuda et al, 1998;Butler et al, 1998). The net e ect of TIMP on tumorigenesis may depend on bioavailability of local amount of TIMPs in tumor microenvironment, the time when the TIMP is presented to the tumor cells, and the presence of putative TIMP receptor on tumor cells.…”

“…In this regard, higher levels of TIMPs may have tumor suppressing e ect, due to its dominant anti-MMP activity, while lower levels of TIMP may favor tumor growth, due to its anti-apoptotic activity. In addition, lower levels of TIMP-2 may also favor proMMP-2 activation by MT1-MMP thereby favoring tumor progression (Shofuda et al, 1998;Butler et al, 1998). Many factors may a ect the activities of TIMPs in tumor.…”

MT1-MMP promotes cell growth and ERK activation through c-Src and paxillin in three-dimensional collagen matrix

“…Human squamous cell carcinoma HSC-4 cells expressed endogenous MT1-MMP [18], and thereby induced the activation of MMP-2, which was stimulated by 3-D collagen culture, and was blocked by MMP inhibitor (BB94) treatment (Fig. 1A).…”