Identification of differentially expressed genes in normal and malignant prostate by electronic profiling of expressed sequence tags.
Differentially expressed genes between corresponding normal and cancertissue can advance our understanding of the molecular basis of malignancy and potentially serve as biomarkers or prognostic markers of malignancy. To identify differentially expressed genes in prostate cancer, we used a procedure combining electronic expression profiling of the prostate expressed sequence tag (EST) database and molecular biology techniques. A novel electronic expression-profiling algorithm was developed to search publicly available EST sequences for genes that show significant differential expression in prostate cancer compared with normal prostate tissue. Approximately 600 genes expressed in prostate were identified through adequate EST counts of ESTs for electronic profiling. Of these 600 genes, 9 showed statistically significant differences in their EST counts between cancer and normal prostate and were further analyzed. The predictions associated with electronic profiling were experimentally verified for two genes, cysteine-rich secretory protein 3 (CRISP-3) and deadenylating nuclease (DAN), using real-time reverse transcription-PCR with total RNA extracted from cells isolated by laser capture microdissection. In five of five Gleason score 6 cancer cases, CRISP-3 expression was increased >50 fold, whereas the expression of DAN was reduced by >80%.
Asmann
Yan W
YW
Division of Experimental Pathology, Department of Laboratory Medicine and Pathology, Mayo Clinic and Foundation, 200 First Street SW, Rochester, MN 55905, USA.
Kosari
Farhad
F
Wang
Kai
K
Cheville
John C
JC
Vasmatzis
George
G
D000465:Algorithms; D018797:Cell Cycle Proteins; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D011467:Prostate / Q000378:metabolism / Q000502:physiology; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012471:Salivary Proteins and Peptides / Q000096:biosynthesis / Q000235:genetics; D029589:Seminal Plasma Proteins / Q000096:biosynthesis / Q000235:genetics
C489926:CRISP3 protein, human; D018797:Cell Cycle Proteins; C490087:NBL1 protein, human; D011506:Proteins; D012471:Salivary Proteins and Peptides; D029589:Seminal Plasma Proteins