Flow cytometric quantitation of the c-myc oncoprotein in archival neoplastic biopsies of the colon.
The c-myc oncogene encoded protein product, p62c-myc, was assayed simultaneously with DNA in populations of individual nuclei extracted from archival biopsies of colonic neoplasia. Both the protein and DNA were assayed fluorimetrically using flow cytometry with a synthetic peptide induced monoclonal antibody (MYC 1-6E10) for the protein and propidium iodide for DNA. The nuclear p62c-myc levels increased progressively from normal mucosa through polyps to carcinomas. However, there was a trend for the more poorly differentiated carcinomas to exhibit lower levels than moderately and well-differentiated tumours, p = 0.085. These results agree with those published previously with the same antibody using Western blotting for protein extracted from fresh frozen tissue and immunocytochemical assessment. Furthermore, flow cytometry is able to effect discriminations between subsets in heterogeneous populations using DNA as a second parameter which Western blot bulk studies cannot.
Watson
J V
JV
MRC Clinical Oncology Unit, Medical Research Council Centre, Cambridge, UK.
Stewart
J
J
Cox
H
H
Sikora
K
K
Evan
G I
GI
D001706:Biopsy; D002457:Cell Extracts / Q000032:analysis; D002467:Cell Nucleus / Q000032:analysis; D003110:Colonic Neoplasms / Q000032:analysis* / Q000473:pathology / Q000648:ultrastructure; D005434:Flow Cytometry; D006801:Humans; D011518:Proto-Oncogene Proteins / Q000032:analysis*; D016271:Proto-Oncogene Proteins c-myc
D002457:Cell Extracts; D011518:Proto-Oncogene Proteins; D016271:Proto-Oncogene Proteins c-myc