High-throughput tissue microarray analysis of cyclin E gene amplification and overexpression in urinary bladder cancer.
Studies by comparative genomic hybridization revealed that the 19q13 chromosomal region is frequently amplified in bladder cancer. The cyclin E gene (CCNE), coding for a regulatory subunit of cyclin-dependent kinase 2, has been mapped to 19q13. To investigate the role of cyclin E alterations in bladder cancer, a tissue microarray of 2,317 specimens from 1,842 bladder cancer patients was constructed and analyzed for CCNE amplification by fluorescence in situ hybridization and for cyclin-E protein overexpression by immunohistochemistry. Fluorescence in situ hybridization analysis showed amplification in only 30 of the 1,561 evaluable tumors (1.9%). Amplification was significantly associated with stage and grade (P: < 0.0005 each). Immunohistochemically detectable cyclin E expression was strong in 233 (12.4%), weak in 354 (18.9%), and negative in 1, 286 of the 1,873 interpretable tumors. The majority (62.1%) of CCNE-amplified tumors were strongly immunohistochemistry-positive (P: < 0.0001). The frequency of protein expression increased from stage pTa (22.2%) to pT1 (45.5%; P: < 0.0001) but then decreased for stage pT2-4 (29.4%; P: < 0.0001 for pT1 versus pT2-4). Low cyclin E expression was associated with poor overall survival in all patients (P: < 0.0001), but had no prognostic impact independent of stage. It is concluded that cyclin E overexpression is characteristic to a subset of bladder carcinomas, especially at the stage of early invasion. This analysis of the prognostic impact of CCNE gene amplification and protein expression in >1,500 arrayed bladder cancers was accomplished in a period of 2 weeks, illustrating how the tissue microarray technology remarkably facilitates the evaluation of the clinical relevance of molecular alterations in cancer.
Richter
J
J
Institute for Pathology and Urologic Clinics, University of Basel, Basel, Switzerland.
Wagner
U
U
Kononen
J
J
Fijan
A
A
Bruderer
J
J
Schmid
U
U
Ackermann
D
D
Maurer
R
R
Alund
G
G
Knönagel
H
H
Rist
M
M
Wilber
K
K
Anabitarte
M
M
Hering
F
F
Hardmeier
T
T
Schönenberger
A
A
Flury
R
R
Jäger
P
P
Fehr
J L
JL
Schraml
P
P
Moch
H
H
Mihatsch
M J
MJ
Gasser
T
T
Kallioniemi
O P
OP
Sauter
G
G
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D019927:Cyclin E / Q000096:biosynthesis / Q000235:genetics*; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D005500:Follow-Up Studies; D005784:Gene Amplification; D015870:Gene Expression; D006801:Humans; D007124:Immunoenzyme Techniques; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology
D019927:Cyclin E; D004273:DNA, Neoplasm; D009363:Neoplasm Proteins