LOH of PTCH1 region in BCC and ovarian carcinoma: microsatellite vs. HRM analysis.
Loss of heterozygosity (LOH) of tumor suppressor genes is a frequent event in tumorigenesis. LOH is most often analyzed by microsatellite typing, but here we offer a fast and efficient method for simultaneous SNP genotyping and mutation scanning, which can also be used for LOH detection. High resolution melting (HRM) provides simple variant detection, and can be adopted for a wide range of applications. When a melting profile for a specific SNP is determined, the screening can be done without the need for sequencing, and only the melting profiles differing from the established melting profiles should be sequenced. LOH of PTCH1 gene is often found in a series of different tumors, for example basal cell carcinoma (BCC) and ovarian carcinoma (OC). In this study LOH was detected in 50 % of BCC and 27.27 % of OC, and the detection rates of microsatellite typing and HRM were comparable. Both methods depended only on the heterozygosity of the loci analyzed, but HRM offers an additional advantage of detection of all sequence variants in the gene of interest.
Division of Molecular Medicine, Rudjer Boskovic Institute, Zagreb, Croatia.
Vesna
V
Musani
Maja
M
Sabol
Diana
D
Car
Petar
P
Ozretic
Slavko
S
Oreskovic
Dinko
D
Leovic
Sonja
S
Levanat
D001483:Base Sequence; D002280:Basal Cell Carcinoma / Q000235:genetics*; D017931:DNA Primers; D005260:Female; D006801:Humans; D019656:Loss of Heterozygosity; D018895:Microsatellite Repeats / Q000235:genetics*; D010051:Ovarian Neoplasms / Q000235:genetics*; D000072079:Patched Receptors; D000072081:Patched-1 Receptor; D011956:Receptors, Cell Surface / Q000235:genetics*; D012878:Skin Neoplasms / Q000235:genetics*
D017931:DNA Primers; C000606236:PTCH1 protein, human; D000072079:Patched Receptors; D000072081:Patched-1 Receptor; D011956:Receptors, Cell Surface