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Performance of Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) on Ethanol Fixed Microscopically Dissected Colonic Aberrant Crypt Foci (ACF)

Published online by Cambridge University Press:  02 July 2020

C.M. Lasko ,
D. Salo ,
C.K. Good  and
R.P. Bird
C.M. Lasko
Affiliation:
Institute of Pathology, Case Western Reserve University, Cleveland, Ohio44106
D. Salo
Affiliation:
Department of Foods and Nutrition, University of Manitoba, Winnipeg, CanadaR3T 2Nz
C.K. Good
Affiliation:
Department of Foods and Nutrition, University of Manitoba, Winnipeg, CanadaR3T 2Nz
R.P. Bird
Affiliation:
Department of Foods and Nutrition, University of Manitoba, Winnipeg, CanadaR3T 2Nz
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Extract

Identification of altered gene expression in colonie preneoplastic lesions may play a key role in early detection and prevention of colon cancer. Considering carcinogen treated rat colonie mucosa harbors a mixed population of both normal and aberrant crypts, it is of interest to independently examine the genetic profile of these two populations. The present study was designed to use a novel method whereby RT-PCR was performed on alcohol fixed ACF which were micro dissected from normal surrounding crypts.

Male F344 rats were injected with azoxymethane, a colon specific carcinogen, at a dose of 15 mg/kg body weight once weekly for 3 weeks. After 28 weeks colons were excised, flushed with RNAse free ice cold phosphate-buffered saline, fixed flat on filter paper, fixed in 70% ethanol and immediately frozen at -80°C After a minimum 24 hours colons were placed on the lid of a sterile, clear petri dish filled with ice.

Type
Neoplasia: Abnormal Cell Growth Or Death/Apoptosis? Insights From Microscopy
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 27 - 28
Copyright
Copyright © Microscopy Society of America 1997

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References

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4.This research was funded by the American Institute for Cancer Research and the Natural Sciences and Engineering Research Council of Canada.Google Scholar