Role of the 21-kDa protein TIMP-3 in oncogenic transformation of cultured chicken embryo fibroblasts.

Abstract

The 21-kDa protein is an extracellular matrix (ECM) component whose synthesis is stimulated transiently during oncogenic transformation ofchicken embryo fibroblasts (CEF) or after treatment of normal cells with the tumor promoter phorbol 12-myristate 13-acetate. Biochemical characterization indicates that the protein is related, but not identical, to two members of the family of tissue inhibitors of metalloproteinases, TIMP-1 and TIMP-2. The cDNA of the 21-kDa protein was recently cloned, and based upon its deduced amino acid sequence and other supporting data we propose that it is another member of this family, a TIMP-3. We now report electrophoretic purification of sufficient quantities of this protein to determine its function. The protein promotes the detachment of transforming cells from the ECM. Although its presence in the matrix may be necessary for cell release it is not the only factor involved because it does not influence the adhesive properties of nontransformed cells. It also appears to accelerate the morphological changes associated with cell transformation and stimulates the proliferation of growth-retarded, nontransformed cells maintained under low serum conditions. Based on these data we hypothesize that the 21-kDa protein promotes the development of the transformed phenotype in cultured cells.The extracellular matrix (ECM) is a dynamic tissue compartment that functions in the regulation of morphogenesis, differentiation (1), and cell proliferation (2). One of its primary functions is proposed to be the sequestration of growth factors (3) that may be released locally in response to physiological stimuli (4). Cell-ECM interactions also play an important role in diseases involving abnormal growth and development, such as cancer, which is characterized by alterations in the synthesis and degradation of matrix components (5).We have been studying the potential roles of various ECM components in oncogenic transformation and have reported characterization of the 21-kDa protein found concentrated in the ECM of transforming chicken embryo fibroblasts (CEF) (6, 7). Synthesis of this protein (Mr 21,000) is stimulated early in the transformation of cells infected with temperaturesensitive mutants of Rous sarcoma virus (RSV) or by treatment of normal, uninfected cells with the tumor promoter phorbol 12-myristate 13-acetate. These observations implicated the 21-kDa protein in the development of the transformed phenotype but did not reveal its precise function.The NH2-terminal amino acid sequence of purified 21-kDa protein is >60% identical to a consensus sequence of mammalian tissue inhibitors of metalloproteinases (TIMPs) and the protein displays metalloproteinase inhibitor activity (8). It is the major inhibitor in the ECM and its solubility properties appear unique among inhibitors with a TIMP-like sequence. Its cDNA was recently cloned and sequenced (9), and its deduced amino acid sequence indicates that it is related to, but distinct from, TIMP-1 and TIMP-2. Based on these and other supporting ...

An immunohistochemical study of TIMP-3 expression in oesophageal squamous cell carcinoma

“…For example, TIMP-1 and TIMP-2 have erythroid potentiating activity, and accelerate the growth of most cells (Hayakawa et al, 1992(Hayakawa et al, , 1994. Tissue inhibitor of metalloproteinase-3 has been reported to stimulate the proliferation of growth-retarded, nontransformed cells maintained under low-serum conditions (Yang and Hawkes, 1992). These reports do not coincide with our data, but growth activity is not always consistent with invasive activity.…”

Gene encoding a novel murine tissue inhibitor of metalloproteinases (TIMP), TIMP‐3, is expressed in developing mouse epithelia, cartilage, and muscle, and is located on mouse chromosome 10

“…TIMP-3 has been shown to be involved in cell transformation Hawkes, 1983,1984;Staskus et al, 1991;Pavloff et al, 1992;Yang and Hawkes, 1992). We have shown here that the Timp-3 gene may also play a role in mouse development.…”

“…It was not present in the culture medium of transformed fibroblasts or in the substratum and medium of untransformed fibroblasts Hawkes, 1983, 1984). ChIMP-3 inhibits matrix metalloproteinases (Staskus et al, 1991;Pavloff et al, 1992), but it also promotes the development of the transformed phenotype, enhancing the loss of cell-ECM adhesion and morphological alterations evident in transforming cultures (Yang and Hawkes, 1992). ChIMP-3 differs from other TIMPs in that it is a component of the ECM (Blenis and Hawkes, 1984).…”

The Activity of Matrix Metalloproteinases (MMPs) and Tissue Inhibitors of Metalloproteinases (TIMPs) in Mammary Tumors of Dogs and Rats

“…In the present study, diffuse in situ gelatinolytic activities were detected in both malignant and benign rat tumor cases, in spite of low MMPs activities, indicating that other gelatinolytic enzymes might play a role. TIMPs are known to have an activity to promote cell growth [3,14,17,27], as well as to inhibit MMPs activity. Therefore, higher activities of TIMPs detected in the rat malignant mammary tumors may contribute to both MMP inhibition and cell growth.…”