ISSN:
1573-2568
Schlagwort(e):
RNA
;
STOOL
;
REVERSE TRANSCRIPTION-POLYMERASE CHAINREACTION
;
GENE EXPRESSION
;
COLON CANCER
Quelle:
Springer Online Journal Archives 1860-2000
Thema:
Medizin
Notizen:
Abstract While colonoscopy may detect early-stage colontumors, a less invasive and more costeffective techniquewould be beneficial. Stool, which picks up sloughed-offcolonic epithelial cells, would be ideal for sampling the mucosa; shed tumor cells maydisplay alterations in gene expression observed inintact tumors. It is first necessary, however, to showthat RNA can be isolated from human feces and that this RNA contains human gene transcripts. We havetherefore developed a method for the isolation of totalRNA from freshly passed human stool, consisting of lysisin chaotropic agents, repeated extraction with phenol and phenolchloroform, and absorptionwith an RNA-binding resin. After treatment withRNase-free DNase I, we assayed these preparations forthe presence of human RNA by quantitative slot blotting, northern blotting, and reversetranscription-polymerase chain reaction (RT-PCR). Weobtained 5-30 μg RNA per gram of stool from cancerpatients, and about 5 μg RNA per gram of controlstool. Quantitative slot blotting showed that about 10% of this RNAwas of human origin. Both northern blotting and RT-PCRdemonstrated the presence of human RNA in these samples.To unambiguously demonstrate the isolation of RNA from stool, we incubated a mixture ofrat cells and control human stool at 37°C for up to24 hr. RT-PCR of the RNA isolated from this sampleclearly revealed the presence of rat-specific mRNA.These experiments indicate that RNA can be isolatedfrom human stool and that message encoded by human genescan be assayed in these preparations. This procedure mayprovide a powerful tool to identify patients at risk for colon cancer.
Materialart:
Digitale Medien
URL:
http://dx.doi.org/10.1023/A:1026699126899
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