Mutations spanning P53 exons 5-9 detected by non-isotopic RNAse cleavage assay and protein expression in human colon cancer.
The non-isotopic assay (NIRCA), based on the observation that RNAse is able to specifically cleave a single mismatch in RNA/RNA duplexes, has been recently proposed to detect p53 mutations. To verify the use of this method as a valid screening for P53 mutations in a routinely collected cancer series, we used this assay on 3 cases with normal and 5 cases with abnormal P53 expression detected by Western blots. In all cases, P53 exons 5-6, 7 and 8-9 regions were analyzed. There were mutations only in the five overexpressed cases: two cases showed mutations in exon 5, one between intron 6 and exon 6 and two in the region spanning exons 8 and 9. Our experience showed NIRCA to be fast, reliable and providing the ability to study long target regions in a single step, thus making this assay useful for genetic screenings.
Tommasi
S
S
National Cancer Institute, Experimental Clinical Oncology Laboratory, Via Amendola, 209-70126 Bari, Italy.
Abatangelo
M
M
Lacalamita
R
R
Montemurro
S
S
Marzullo
F
F
Paradiso
A
A
D003110:Colonic Neoplasms / Q000235:genetics*; D005091:Exons; D016158:Genes, p53; D006801:Humans; D009154:Mutation; D012260:Ribonucleases / Q000378:metabolism*
D012260:Ribonucleases