Inhibition of metastasis in human gastric cancer cells transfected with tissue inhibitor of metalloproteinase 1 gene in nude mice

Abstract

BACKGROUND. Tissue inhibitors of nietalloproteinases (TIMPs) act as negative regulators of matrix metalloproteinases (MMPs) that degrade extracellular matrix. We evaluated the metastatic ability of the highly metastatic human gastric cell line KKLS, and that of cells transfected with exogenous TIMP-I gene by the orthotopic transplantation model in nude mice. METHODS. KKLS was derived from human gastric cancer. Expression of mRNA for tissue inhibitor of nietalloproteinase-1 (TIMP-1) was almost undetectable in KKLS cells. KKLS cells were transfected with exogenous TIMP-1 gene by the Chen-Okayama method. Two clones (KTCLs) that expressed different levels of TIMP-1 and neomycin-resistant KKLS (K-neo) were obtained. The KKLS cells and these transfectants were orthotopically transplanted into nude mice (murine stomach) and metastasis in the murine liver was detected. As a method of detecting metastasis, we used a DNA fragment (human P-globin gene) specific to human tumor cells that have metastasized into the murine liver by polymerase chain reaction (PCR). RESULTS. Differences in tumor growth in the murine stomach were not observed between KKLS cells, K-neo cells, and the two transfectants expressing the different TIMP-1 levels (low, high,. The KKLS cells and K-neo cells had undergone liver metastasis, as shown by PCR amplification of the human ,&globin gene fragment from the murine liver samples, since Week 1 after transplantation and the metastasis had grown exponentially; however, although KTCL-1 cells and KTCL-14 cells had undergone liver metastasis since Week 2, the metastasis had not grown. The average intensities of the amplified gene fragments from K-neo cells, KTCL-1 cells, and KTCL-14 cells in Week 4 after transplantation were loo%, 45%, and 18%, respectively, of the parental KKLS cells. CONCLUSIONS. TIMP-1 was suggested to act as a negative regulator of the nietastasis. The present data is thought to be especially important because the mice in this study underwent orthotopic transplantation with a metastatic model. Cancer 1996; 77:1676-80.

Runx3 suppresses gastric cancer metastasis through inactivation of MMP9 by upregulation of TIMP‐1

“…In primary prostate cancer cell lines, TIMP1 expression induced by IL10 significantly blocked tumor cell invasion 47. In the orthotopic transplantation model in nude mice, metastatic ability of the highly metastatic human gastric cell line KKLS is significantly decreased by exogenous TIMP‐1 gene transfection 48. Western blot, semiquantitative RT‐PCR and ELISA showed that TIMP1 expression was elevated by RUNX3 in gastric cancer cells both at the mRNA and protein level, and this may contribute to the reduced MMP9 enzyme activity seen when RUNX3 is exogenously expressed.…”

Adenovirus-Mediated Overexpression of Tissue Inhibitor of Metalloproteinase-1 Reduces Atherosclerotic Lesions in Apolipoprotein E–Deficient Mice

“…Human TIMP-1 inhibited murine MMPs in vitro (Figure 4), a finding consistent with previous work showing that murine MMPs were inhibited in transgenic mice overexpressing human TIMP-1. 32,33 In addition, MMP-3, MMP-13 (Figure 7, C through F), and MMP-2 (data not shown) have been immunolocalized in lesions of our apoE Ϫ/Ϫ mice: one consequence of MMP inhibition might be the limitation of the proteolysis of elastin, collagen, and ␣-actin ( Figure 6, A through F). Indeed, inhibition of MMPs by local overexpression of TIMP-1 increases elastin accumulation in rat carotid artery intima.…”

Matrix metalloproteinases in tumour invasion and metastasis

“…An anti-metastatic effect for TIMP-1 in gastric cancer has been reported, 66 but more recent immunohistochemical studies have not observed a correlation between TIMP-1 presence and tumour progression. The successful production of monoclonal antibodies to TIMP-1 and TIMP-2 has recently been described.…”