Multiparametric in situ mRNA Hybridization Analysis of Gastric Biopsies Predicts Lymph Node Metastasis in Patients with Gastric Carcinoma

Abstract

We examined the expression level of several genes that regulate different steps in metastasis formation in formalin-fixed, paraffin-embedded biopsies of 189 primary human gastric carcinomas prior to surgical resection in patients in whom lymph node metastasis was not evident by endoscopic ultrasound or computed tomography (CT) scan. The expressions of epidermal growth factor receptor (EGF-R), vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2 and E-cadherin were examined by a colorimetric in situ mRNA hybridization technique. The integrity of the mRNAs was verified, leaving 161 (85.2%) patients for study. After gastrectomy, 82 patients had positive lymph nodes and 79 patients had negative lymph nodes. The concurrent expression levels of MMP-2 and E-cadherin mRNAs were significantly higher and lower, respectively, in the metastatic tumors than the non-metastatic tumors. Expression of EGF-R and VEGF was not different between the metastatic and non-metastatic tumors. However, when only the intestinal-type of gastric cancer was evaluated, the level of VEGF mRNA was significantly higher in tumors associated with lymph node metastasis than in those without metastasis. However, a high MMP-2:Ecadherin ratio significantly correlated with lymph node metastasis in both types of gastric cancer. These results suggest that multiparametric in situ hybridization analysis for several metastasisrelated genes may allow the preoperative prediction of lymph node metastasis from gastric cancer. Key words:Metastasis-related genes -Gastric carcinoma -Biopsy specimen -Lymph node metastasis -in situ mRNA hybridizationThe prognosis and choice of therapy for gastric cancer patients are based on the stage of the disease as well as its metastatic potential. 1) However, routine histopathological examination of primary gastric cancer specimens cannot always identify patients with metastasis, especially lymph node metastasis. Recent advances in molecular biology and in our understanding of the pathogenesis of metastasis have provided new tools with which to predict the metastatic potential of human cancers.To produce a metastasis, tumor cells must complete all the steps in the metastatic cascade, including transformation, growth, angiogenesis, invasion, survival in the circulation, adhesion, extravasation, proliferation, and again angiogenesis at the site of metastasis.2) Numerous reports have demonstrated that in different tumor systems the metastatic potential of cells directly correlates with the expression of different genes that encode for epidermal growth factor receptor (EGF-R), [3][4][5][6][7][8] basic fibroblast growth factor (bFGF), 9-12) vascular endothelial growth factor (VEGF), [13][14][15][16] interleukin-8, 17,18) matrix metalloproteinase (MMP)-2, [19][20][21][22] multidrug resistance (mdr-1) 23) and carcinoembryonic antigen (CEA), 24,25) and inversely with expression of Ecadherin. [26][27][28] Most of these correlative studies reached the inevitable conclusion that the expression of a given gene is neces...

Endoscopic biopsies from gastrointestinal carcinomas and their suitability for molecular analysis: a review of the literature and recommendations for clinical practice and research

“…Among studies investigating ERBB2 copy number by ISH, four used the same criteria to define ERBB2 amplification [ERBB2/chromosome 17 centromere (CEP17) ratio of ≥2.2 in 20 tumour cell nuclei or a ERBB2/CEP17 ratio of ≥2 in 40 tumour cell nuclei where the ratio in 20 nuclei lay between 1.8 and 2.2], whereas all other studies used different criteria . ISH was also used to investigate EGFR copy number in gastric carcinoma . The concordance rate between biopsy and matched resection specimen for EGFR status varied by up to 30% in 80% of patients in one study of 189 patients and was 0% in a much smaller second study of 14 patients …”

“…ISH was also used to investigate EGFR copy number in gastric carcinoma . The concordance rate between biopsy and matched resection specimen for EGFR status varied by up to 30% in 80% of patients in one study of 189 patients and was 0% in a much smaller second study of 14 patients …”

“…6,12,13,48 ISH was also used to investigate EGFR copy number in gastric carcinoma. 6,55 The concordance rate between biopsy and matched resection specimen for EGFR status varied by up to 30% in 80% of patients in one study of *Heterogeneity was defined as the expression of the investigated markers in some but not all the biopsies taken, or in only some regions of the resection specimen. 12 189 patients 55 and was 0% in a much smaller second study of 14 patients.…”

Blockade of NF-κB nuclear translocation results in the inhibition of the invasiveness of human gastric cancer cells

“…MMPs, which are important in the breakdown of the extracellular matrix (ECM), are overexpressed in malignant tumors and have been demonstrated to contribute to tumor proliferation, invasion and metastasis (19). Among the MMPs, MMP-9 has a close association with tumor metastasis, and is considered, in particular, to be an important factor in facilitating invasion and metastasis in gastric carcinoma (20–22). TIMPs (TIMP-1, -2, -3 and -4) have been demonstrated to be the key regulators of MMP activity and ECM degradation (23).…”

Effects of LY294002 on the invasiveness of human gastric cancer in vivo in nude mice

“…Among the MMPs, MMP-2 and MMP-9 are closely related to the metastasis of tumors, and have been specifically considered to be important factors in facilitating lymphatic invasion and metastases in gastric carcinoma [20][21][22] . MVD is a quantitative index for carcinoma angiogenesis, which indicates the regulation of the progress and metastasis of primary gastric carcinoma, and can therefore serve as a marker for carcinoma prognosis [23][24][25] .…”