Analysis Of Differential Expression Profile Of Breast Cancerusing RFDD-PCR

Objective To establish restriction fragment differential display polymerase chain reaction (RFDD-PCR) as an efficient technique for constructing and studying gene expression profile of human tissues. To screen differentially expression gene profile of breast cancer and identify candidate genes. Material & Method collecting tissues of mammary adenocarcinoma(T), cancerometastasis lymph node(L) and normal mammary(N) from one mammary infitrating ductal carcinoma case, gene expression profile of each kind of tissue has been constructed using RFDD-PCR technique at equal pace according to operating manual of Qbio-gene company. Then all fragments of the three gene expression profiles have been separated and displayed by electrophoresis on 7% urea denaturing polyacrylamide gel. With the gene database at the website http://www.Qbio-gene.com/display, we identified names of probable fragments by bioinformatics analysis. Through comparison among the three profiles, the numbers and types of most differentially expressed gene fragments were displayed and screened. Results The expression profile of three kinds of tissue has been constructed covering 1716 fragments of mammary adenocarcinoma, 1769 of cancerometastasis lymph nodes and1922 of normal mammary tissue. Among these 5407 fragments, 39.39% were exactly same. While 33.9% sequences of T and L showed differences in abundance or presence, 40.9% of T and N and 39.6% fragments of L and N were observed differentially expressed. These differentially expressed gene fragments are analyzed and screened, part of them could be new breast caner candidate genes. Fragments which are possible to be novel genes/ESTs were selected too. These fragments are to be studied. Conclusions RPDD-PCR is an efficient technique for research in human diseases genomics as a mass screening to complete gene expression profile with high-flux. Through comparison among three or more profiles, screening of candidate genes of a certain disease can be done, and it has good chance to identify novel gene or EST. it is essential for studying disease involving multi-genes as breast cancer to survey whole gene expression profile and screen all possible candidate genes.