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Diagnostic Technologies and Molecular and Cellular Profiling: Biomarkers in Early Detection and Diagnosis

Mass RNA analysis using stool in colorectal cancer

Ewha Womans University, Seoul, Republic of Korea

Abstract

A29

(Background) It has been estimated that approximately 10¹⁰ of normal adult colonic epithelial cells, each having a lifespan of 3-4days, are shed daily from the lower two-thirds of colon crypts; thus, using colonocytes to develop a screening test is an attainable goal. There is a need for sensitive and specific diagnostic and prognostic molecular markers which can monitor early patterns of gene expression in non-invasive exfoliated colonocytes shed in the stool. There were a few studies of RNA -based detection method because of the characteristics in stool. We report here the mass RNA analysis using stool in colorectal cancer. (Method) The study included 15 patients with colorectal cancer, and 15 control patients without neoplastic disease of the colon and rectum. RNA was isolated from routinely collected stool samples using a modified method. The expression levels of survivin, livin, Akt-1, caveolin, histone deacetylase(HDAC)1, matrix metalloproteinases(MMP)-2, MMP-7, MMP-9, MMP-12, hepatoma derived growth factor(HDGF), peptideYY, hypoxia-inducible factor(HIF)-1, epidermal growth factor receptor(EGFR), N-cadherin, catenin-ß, inducible nitric oxide synthase(iNOS), ring-3, enolase-1, insulin-like growth factor binding protein(IGFBP)-2, tissue inhibitors of metalloproteinase(TIMP)-1 and EphB2 were determined by reverse-transcription polymerase chain reaction(RT-PCR). (Result) The sensitivity of fecal survivin, livin and Akt-1 assays for colorectal cancer was 93%, whrereas sensitivity of the fecal caveolin, HDAC1, MMP-2, HDGF and peptideYY assays for colorectal cancer was 13%, 6%, 20%, 6% and 6%, respectively. The remained 13assays were not shown any expression in both colorectal and normal group. The expression levels of survivin, livin and Akt-1 were higher in colorectal cancer group than normal group(p<0.05). (Conclusion & Inquiry) This fecal survivin, livin and Akt-1 assay had high sensitivity and high expression levels(colorectal cancer as distinguished from normal group) for detecting colorectal cancer, although a larger study is necessary to asses the sensitivity and expression level.