Application of restriction fragment differential display-polymerase chain reaction in study on differential expression profiles of human diseases.
OBJECTIVE
To establish the restriction fragment differential display-polymerase chain reaction (RFDD-PCR) as an efficient technique for constructing and studying the gene expression profile of human tissues.


METHODS
The tissues of mamma adenocarcinoma (T), cancerometastasis lymph node (L) and normal mammary (N) from one mammary infiltrating ductal carcinoma case were collected, and the gene expression profile of each kind of tissue was constructed using RFDD-PCR technique at equal pace according to the operating manual of Qbio-gene Company. Then all fragments of the three gene expression profiles were separated and displayed by electrophoresis. With the use of gene database at the website http://www.Qbio-gene.com/display, the authors identified the names of the probable fragments by bioinformatics analysis. Through comparison of the three profiles, the numbers and types of most differentially expressed gene fragments were displayed.


RESULTS
The expression profiles of the three kinds of tissue have been constructed covering 1716 fragments of mammary adenocarcinoma, 1769 of cancerometastasis lymph nodes and 1922 of normal mammary tissue. Among these 5407 fragments, 39.39% were exactly the 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 were found to relate with metastasis, differentiation, inflammation and so on.


CONCLUSIONS
RFDD-PCR is an efficient technique for research in human diseases genomics as a mass screening for complete gene expression profile with high-flux. Through comparison among three or more profiles, the screening for candidate genes of a certain disease can be accomplished, and there is probably a chance to identify novel gene or expressed sequence tag.
Zhou
Hong-ying
HY
Department of Anatomy, School of Preclinical & Forensic Medicine, Sichuan University, Chengdu, Sichuan, 610041, PR China.
Mei
Yan
Y
Lu
You-guang
YG
Li
Ai-dong
AD
Tang
En-jie
EJ
Yang
Hui-jun
HJ
D000230:Adenocarcinoma / Q000235:genetics; D001943:Breast Neoplasms / Q000235:genetics; D018270:Carcinoma, Ductal, Breast / Q000235:genetics; D019295:Computational Biology; D004586:Electrophoresis / Q000379:methods; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods