Semi-quantitative fluorescent PCR analysis identifies PRKAA1 on chromosome 5 as a potential candidate cancer gene of cervical cancer.
OBJECTIVE
Comparative genomic hybridization has frequently detected amplification of chromosome 5p in cervical cancer, but candidate cancer genes within the region are rarely known. Therefore, we pursued to identify potential candidate gene related to cervical cancer development.


METHODS
A series of 128 cervical tumor samples were examined by semi-quantitative fluorescent differential PCR for copy number changes on three candidate genes (PRKAA1, CTNND2 and POLS) mapped to chromosome 5p and one gene (ERBIN) mapped to chromosome 5q12.3. The impact of gene copy number was later analyzed in relation to HPV infection, tumor stage or tumor radiosensitivity.


RESULTS
DNA copy numbers of PRKAA1, CTNND2 and ERBIN were significantly different from normal controls (P < 0.05). DNA copy number changes did not correlate with HPV infection, tumor stages or tumor radiosensitivity. Using RT-PCR, PRKAA1 mRNA expression in seven tumor samples with known 5p amplification was amplified from 3- to 15-fold. Over-expression of PRKAA1 was further confirmed by immunohistochemical staining on 125 paraffin-embedded cervical cancer tissues. The expression level in cervical tumor was significantly higher than that in normal epithelium (P < 0.001).


CONCLUSIONS
PRKAA1 gene codes for the catalytic alpha 1 subunit of the AMP-activated protein kinase which is an important cellular metabolic stress regulator. It might assist tumor cells growth under stress. Thus, PRKAA1 may be one of the potential candidate genes for cervical carcinogenesis.
Huang
Fung Yu
FY
Department of Obstetrics and Gynaecology, Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China.
Chiu
Pui Man
PM
Tam
Kar Fai
KF
Kwok
Yvonne K Y
YK
Lau
Elizabeth T
ET
Tang
Mary H Y
MH
Ng
Tong Yow
TY
Liu
Vincent W S
VW
Cheung
Annie N Y
AN
Ngan
Hextan Y S
HY
D055372:AMP-Activated Protein Kinases; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D000328:Adult; D000368:Aged; D002294:Carcinoma, Squamous Cell / Q000235:genetics*; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics; D002895:Chromosomes, Human, Pair 5 / Q000235:genetics*; D004273:DNA, Neoplasm / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D005260:Female; D005784:Gene Amplification; D018628:Gene Dosage; D006801:Humans; D008875:Middle Aged; D009097:Multienzyme Complexes / Q000235:genetics*; D009687:Nuclear Proteins / Q000235:genetics; D027383:Papillomaviridae / Q000235:genetics; D030361:Papillomavirus Infections / Q000150:complications / Q000201:enzymology / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D002583:Uterine Cervical Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000821:virology
D048868:Adaptor Proteins, Signal Transducing; D002868:Chromosomal Proteins, Non-Histone; D004273:DNA, Neoplasm; D004279:DNA, Viral; C413364:ERBIN protein, human; D009097:Multienzyme Complexes; D009687:Nuclear Proteins; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases; C526180:PRKAA1 protein, human; D004259:DNA-Directed DNA Polymerase; C118146:TENT4A protein, human