Expression of reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) as a prognostic indicator in gastric cancer

https://doi.org/10.1016/j.ejca.2005.09.016Get rights and content

Abstract

In this study the expression levels of reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) in gastric cancer cell lines and tissues have been analysed in order to assess their value as a prognostic indicator. The expressions of RECK, activated matrix metalloproteinase (MMP)-7, and vascular endothelial growth factor (VEGF) in gastric cancer tissues and cell lines were evaluated by Western blot analysis; and MMP-2 and MMP-9 were evaluated by gelatin zymography. RECK expression in the context of gastric cancer was also compared with various clinicopathologic parameters and compared to the expression of activated MMP-7, MMP-2, and MMP-9. Fifty-two percent of the 102 gastric cancer tissues and 81.8% of the 11 gastric cancer cell lines exhibited reduced RECK expression. We also detected a significant inverse correlation between RECK expression and macroscopic tumour growth (P = 0.018), lymphatic invasion (P = 0.018), lymph node metastasis (P = 0.000), stage (P = 0.000), and MMP-9 (P = 0.039). No correlation between RECK expression and MMP-7 and MMP-2, VEGF were detected. Our data strongly supports the hypothesis that RECK is a suppressor of malignancy, and constitutes a good prognostic indicator in gastric cancer.

Introduction

The gene for reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) was initially isolated using an expression cloning strategy, which was designed to detect transformation-suppressor genes against activated ras oncogenes [1]. Also, the RECK gene is expressed widely in a variety of normal human tissues and non-neoplastic cell lines, and its expression is either low or undetectable in oncogene-transformed fibroblasts and many tumour-derived cell lines 2, 3. Previous experimental studies have revealed that RECK is able to inhibit tumour angiogenesis, invasion, and metastasis 2, 3, 4, 5. It also inhibits matrix metalloproteinase (MMP)-9, MMP-2, and MT1-MMP (MMP-14) secretion and activity, via an as-yet-unknown mechanism 2, 4. Vascular endothelial growth factor (VEGF) is considered a key mediator of tumour angiogenesis, including neovascularization in human cancer. Several VEGF isoforms are produced from a single gene as a result of alternative splicing [6]. The isoforms differ in their biological properties and in their abilities to bind heparan sulfate proteoglycans [7]. VEGFs are bioactive as freely diffusible proteins in the extracellular space where they become available to endothelial cells, and in one report it has been suggested that soluble isoforms VEGF 121 or VEGF 165 have greater angiogenic and tumourigenic properties than the heparin-bound form [8]. In support of this hypothesis, RECK expression in a variety of human tumours, as compared with normal tissues, was correlated with prolonged survival in patients suffering from hepatocellular carcinoma [9], pancreatic cancer [10], breast cancer [11], non-small cell lung cancer [12], and colorectal cancer [13].

Gastric cancer is the second most common cause of cancer death in the world, and is also one of the most common cancers in East Asia and South America, although its incidence in Western Europe and North America is now declining. Most patients who are diagnosed with gastric cancer exhibit advanced disease, and prognoses are extremely poor, with survival rates rarely exceeding 50%. However, patients suffering from cancers which are limited to the mucosae and submucosae, namely early gastric cancer (EGC), have a five-year survival rate of approximately 95%. Therefore, the management of gastric cancer is intimately dependent on tumour stage. In addition, lymph node metastasis is also very important, even in patients with EGC. Five-year survival rates of the node negative and positive EGC patients are significantly different, with a 93–99% survival rate for the node-negative patients, and a 73–90% survival rate for the node-positive patients [14]. It is therefore important to delineate a novel set of parameters which can be used to target lymph node metastasis and staging.

In this study, we have analysed RECK expression levels in gastric cancer cell lines and tissues, in order to assess their status in gastric cancer tissues, and their value as prognostic indicators. We have also analysed MMP-7 activation, and compared the data with those for RECK expression. Our results indicate that the level of RECK expression in gastric cancers correlates inversely with certain metastasis-related clinicopathological and molecular features.

Section snippets

Cell lines and tissue samples

The following human gastric caner cell lines were studied: SNU1, SNU5, SNU16, SNU620, SNU601, SNU638, SNU668, M1, M28, M45, and M74. All cancer cell lines were cultured in RPMI 1640, supplemented with 10% FBS, 1 mM NaCO3, 2 mM l-glutamine, penicillin–streptomycin, in a 5% CO2 atmosphere, at 37 °C. 102 patients with gastric adenocarcinoma, who had undergone curative surgery (total or subtotal gastrectomy with tumour-free resection margins, with regional lymphadenectomy), were included in this

RECK expression in gastric cancer cell lines and tissues

Of the 11 gastric cancer cell lines examined, nine (81.8%) did not show RECK protein expression by western blot analysis (Fig. 1A). Only two cell lines, S16 and M74, exhibited RECK protein expression. Of the 102 gastric cancer tissues, 53 (52.0%) displayed a reduced expression of RECK protein (Fig. 1B).

General clinicopathologic characteristics of studied cases

The 102 patients studied included 61 males (59.8%) and 41 females (40%), and ranged in age from 30 to 78 years, with a mean age of 57.7 years. Tumour size ranged from 3.0 to 17.0 cm, with a mean

Discussion

The inverse relationship between RECK expression in various human tumours and their prognoses has been amply demonstrated in patients suffering from hepatocellular carcinoma [9], pancreatic cancer [10], breast cancer [11], non-small cell lung cancer [12], and colorectal cancer [13]. However, the significance of RECK expression, not only in gastric cancer but also in non-cancerous gastric tissue, has yet to be elucidated and warrants pilot studies in other types of human tumours [12]. In this

Conflict of interest statement

None declared.

Acknowledgement

This work was supported by the Samsung Biomedical Research Institute grant, #SBRI C-A5-114-1.

References (27)

  • G. Neufeld et al.

    Vascular endothelial growth factor (VEGF) and its receptors

    FASEB J

    (1999)
  • H.T. Zhang et al.

    The 121 amino acid isoform of vascular endothelial growth factor is more strongly tumourigenic than other splice variants in vivo

    Br J Cancer

    (2000)
  • T. Masui et al.

    RECK expression in pancreatic cancer: its correlation with lower invasiveness and better prognosis

    Clin Cancer Res

    (2003)
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