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Complementary DNA (cDNA) microarrays were applied to identify aberrantly expressed genes in patients with gastric cancer. Among the most exuberantly expressed genes Glyoxalase I (GLO1) was one that encodes. We assessed whether overexpression of GLO1 was a potential risk factor for gastric cancer. GLO1 mRNA transcripts and protein expression levels were determined by Western blot test and real time quantitative PCR (qPCR). These tests verified that the expression of GLO1 was higher in gastric cancer tissues compared to adjacent noncancerous tissues. Furthermore, Immunohistochemistry were studied in paraffin-fixed sections of gastrectomized specimens of 114 patients. The IHC scores of GLO1 in patients with serosa invasion (T3 and T4) were significantly higher than those without serosa invasion (T1 and T2). Also, the IHC scores of GLO1 were significantly greater in patients with lymph node metastasis than those without metastasis, and were significantly higher in stages III and IV than in stages I and II. Five-year survival of patients with lower expression of GLO1 gene was significantly better than among patients with a higher expression. To investigate the functional significance of GLO1 expression, GLO1 short hairpin RNA (shRNA) expression plasmids were transfected into gastric cancer cell lines. Cancer cells depletion of GLO1 significantly reduced the migration and invasion ability of each cell line. The results point to GLO1 as prospective prognostic marker for gastric cancer and its role in gastric cancer progression.Key words: gastric cancer, molecular biomarker, prognostic factor, GLO1