Mutations Detection Of Emsy In Sporadic Breast Cancer With PCR-SSCP

Objective: The purpose of this study was to determine whether EMSY gene (C11orf30 in genebank) mutation is present in breast cancer tissue and investigate the role of EMSY gene in human breast tumorigenesis.Methods: Genomic DNA was isolated from 72 sporadic breast cancer tissue and 18 benign breast tumor tissue using the Phenol-Chloroform extraction method. Polymerase chain reaction (PCR) was used to amplify twenty exons of the EMSY gene. The entire EMSY gene coding sequence was amplified by PCR with primers especially designed for mutation screening by single-strand conformation polymorphism(SSCP), and variant bands were sequenced.Results: Among the 72 cases of sporadic breast carcinoma specimens and 18 cases of benign breast tumor specimens, there was an abnormal SSCP electrophoresis band in exon3, and the mutation rate was 1.4%. Through DNA sequencing, the mutation type was missense mutation and the albumen structure was (AAG→CAG, K→Q). Also in exon9, 6 breast carcinoma specimens have conformation change in single stranded. The sequencing result were absence A of GCA, the mutation rate was 8.3%. Further, a novel point mutation in exon20 was detected in all samples (4.2%). Through analysis, there were three synonymous mutations of ACT→ACC (threonine,Thr).Couclusion: The study suggest that EMSY gene mutation is present in sporadic breast cancer in China, but the mutation rate is low. It is likely that there are other important susceptibility genes that remain to be identified with sporadic breast cancer.