Tumor Growth Suppression in Pancreatic Cancer by a Putative Metastasis Suppressor Gene Cap43/NDRG1/Drg-1 through Modulation of Angiogenesis

Department of Surgery, Kurume University, Куруме, Fukuoka, Japan
Cancer Research (Impact Factor: 9.33). 07/2006; 66(12):6233-42. DOI: 10.1158/0008-5472.CAN-06-0183
Source: PubMed


Cap43 has been identified as a nickel- and calcium-induced gene, and is also known as N-myc downstream-regulated gene 1 (NDRG1), Drg-1 and rit42. It is also reported that overexpression of Cap43 suppresses metastasis of some malignancies, but its precise role remains unclear. In this study, we asked how Cap43 could modulate the tumor growth of pancreatic cancer. Stable Cap43 cDNA transfectants of pancreatic cancer cells with Cap43 overexpression showed similar growth rates in culture as their control counterparts with low Cap43 protein level. By contrast, Cap43 overexpression showed a marked decrease in tumor growth rates in vivo. Moreover, a marked reduction in tumor-induced angiogenesis was observed. Gelatinolytic activity by matrix metalloproteinase-9 and invasive ability in Matrigel invasion activity were markedly decreased in pancreatic cancer cell lines with high Cap43 expression. Cellular expression of matrix metalloproteinase-9 and two major angiogenic factors, vascular endothelial growth factor and interleukin-8, were also significantly decreased in cell lines with Cap43 overexpression as compared with their parental counterparts. Immunohistochemical analysis of specimens from 65 patients with pancreatic ductal adenocarcinoma showed a significant association between Cap43 expression and tumor microvascular density (P = 0.0001) as well as depth of invasion (P = 0.0003), histopathologic grading (P = 0.0244), and overall survival rates for patients with pancreatic cancer (P = 0.0062). Thus, Cap43 could play a key role in the angiogenic on- or off-switch of tumor stroma in pancreatic ductal adenocarcinoma.

Download full-text


Available from: Toshiro Yokoyama, Jun 19, 2015
  • Source
    • "It normally presents in cells of human epithelial tissues, including tissues of digestive, urinary, and respiratory tracts, and is regarded as a modulator to regulate tumor cell differentiation , proliferation, invasion, and metastasis; and also, in cell cycle regulation, vesicular transport, and maintenance of membrane E-cadherin [3] [4] [5]. Recent evidence demonstrated that NDRG1 is negatively correlated with tumor progression and acts as a potent metastasis suppressor [4] [6] [7]. Studies showed that NDRG1 inhibited metastasis by reducing cell-matrix and cell-cell adhesion in human cancer cells [8]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Oral squamous cell carcinoma (OSCC) is the most common phenotype of oral cancer. N-myc downstream regulated gene 1 (NDRG1) is a modulator for cell proliferation, differentiation, and invasion. The role and function of NDRG1 in OSCC cells remain inconclusive. The (3)H-thymidine incorporation and in vitro matrigel invasion assays revealed NDRG1-knockdown significantly enhanced OSCC cell proliferation and invasion. Overexpressed NDRG1 arrested the cell cycle at the S-phase, thus attenuated cell proliferation in OECM-1 cells. The NDRG1-knockdown enhanced tumorigenesis of OECM-1 cells in the xenograft animal model. Western-blot and zymographic assays revealed NDRG1 downregulated the gelatinase activities and protein levels of metalloproteinase-2 (MMP-2) and metalloproteinase-9 (MMP-9). NDRG1 modulated epithelial-mesenchymal transition (EMT) through upregulation of the E-cadherin expression, but downregulation of the N-cadherin, Vimentin, Snail-1, and Slug. Immunofluorescence staining indicated knockdown of NDRG1 enhanced F-actin expression and polymerization. Our results indicated NDRG1 attenuated OSCC cell growth in vitro and in vivo. The downregulation of EMT, MMP-2, and MMP-9 may explain the role of anti-invasion of NDRG1 in human OSCC cells. The experiments recognize that NDRG1 is an antitumor gene in OSCC cells.
    Full-text · Article · Nov 2014 · Cancer Letters
  • Source
    • "controversial in different cancer types [11] [12] [13], and NDRG1 had not been investigated in RCC until this study. By immunohistochemistry, we established a correlation between NDRG1 expression and favorable prognosis in the 82 newly enrolled patients. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The aim of this study was to identify proteins with aberrant expression in clear cell renal cell carcinoma (ccRCC), and elucidate their clinical utilities. The protein expression profiles of primary ccRCC tumor tissues and neighboring non-tumor tissues were obtained from 9 patients by two-dimensional difference gel electrophoresis and mass spectrometry. Comparative analysis of 3771 protein spots led to the identification of 73 proteins that were expressed at aberrant levels in tumor tissues compared with non-tumor tissues. Among these 73 proteins, we further focused on N-myc downstream-regulated gene 1 protein (NDRG1). NDRG1 expression is regulated by members of myc family as well as by p53, HIF1A, and SGK1. The biological and clinical significance of NDRG1 is controversial for various malignancies and no detailed studies on NDRG1 have been reported in ccRCC until our study. For the 82 newly enrolled ccRCC patients, immunohistochemical analysis revealed a significant association between nuclear NDRG1 and favorable prognosis (p<0.05). Multivariate analysis demonstrated the role of NDRG1 as an independent factor of progression-free survival (p=0.01). Subsequent in vitro gene suppression assay demonstrated that NDRG1 silencing significantly enhanced cell proliferation and invasion of RCC cells. The cytotoxic effects of the NDRG1 up-regulation induced by iron chelator were also confirmed. These findings suggest that nuclear NDRG1 has tumor suppressive effects, and the NDRG1 expression may have clinical values in ccRCC. Nuclear NDRG1 may provide additional insights on molecular backgrounds of ccRCC progression, and contribute to the development of novel therapeutic strategy.
    Full-text · Article · Aug 2013 · Biochimica et Biophysica Acta (BBA) - Proteins & Proteomics
  • Source
    • "Many studies examining different cell-types suggest that various primary antibodies used to detect NDRG1 recognize both non-phosphorylated and phosphorylated forms, resulting in two distinct bands detected by Western blotting [7,13,15–24]. Two reports have suggested that phosphatase treatment of cell lysates depletes phosphorylated NDRG1 detected by Western-blot analysis in HUVECs (human endothelial umbilical vein endothelial cells) [15] and MIAPaCa-2 pancreatic cancer cells [16]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: N-myc downstream regulated gene-1 (NDRG1) is a metastasis suppressor that is down-regulated in prostate cancer. NDRG1 phosphorylation is associated with inhibition of metastasis and western blots indicate two bands at ~41 and ~46 kDa. Previous investigations by others suggest the higher band is due to NDRG1 phosphorylation. However, the current study using a dephosphorylation assay and the Phos-tag SDS-PAGE assay, demonstrated the 46 kDa NDRG1 protein band was not due to phosphorylation. Further experiments showed the NDRG1 protein bands were not affected upon glycosidase treatment, despite marked effects of these enzymes on the glycosylated protein, fetuin. Analysis using RT-PCR demonstrated only a single amplicon, and thus, the two bands could not a result from an alternatively spliced NDRG1 transcript. Western blot analysis of prostate cancer cell lysates identified the 41 kDa band to be a truncated form of NDRG1, with mass spectrometry confirming the full and truncated proteins to be NDRG1. Significantly, this truncated protein was not present in normal human prostate epithelial cells. Western blot analysis using anti-NDRG1 raised to its N-terminal sequence failed to detect the truncated protein, suggesting it lacked N-terminus amino acids (residues 1-49). Sequence analysis predicted a pseudotrypsin protease cleavage site between Cys49-Gly50. Such cleavage of NDRG1 in cancer cells may result in loss of NDRG1 tumour suppressive activity.
    Full-text · Article · May 2013 · Bioscience Reports
Show more