Analysis of prostate tissue DNA for the presence of human papillomavirus by polymerase chain reaction, cloning, and automated sequencing.
We have analysed the DNA from 24 prostate tissue biopsies, spanning a range of Gleason grading from benign to grade 5 and mixed randomly with cervical cancer samples of known human papillomavirus (HPV) status, for the prevalence of HPV DNA, in a double-blind study to ensure complete objectivity. Polymerase chain reactions (PCR) were performed using general E1 open reading frame primers for HPV under low stringency conditions, in addition to reactions containing primers specific for HPV16, E2, and E6 open reading frames under higher, more stringent PCR conditions. The presence of cellular DNA was verified by the use of primers for hypoxanthine guanine phosphoribosyl transferase. DNA bands were not detected in the prostate biopsies using the HPV16-specific primers under high-stringency PCR conditions, however a predominant band in the 400 bp region was observed in 15 of the prostate biopsies using the general primers and the low annealing temperature of 40 degrees C. This fragment was excised and cloned into the pT7 blue vector and the sequence of the insert determined. Although the cloned sequences initiated and terminated with the two authentic PCR primers, they did not contain a significant HPV-related open reading frame. Our results indicate that HPV type 16 and closely related types, as detected by the general primer pair, are unlikely initiators of prostate carcinogenesis within our population.
Anderson
M
M
Department of Biology, University of York, United Kingdom.
Handley
J
J
Hopwood
L
L
Murant
S
S
Stower
M
M
Maitland
N J
NJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D004268:DNA-Binding Proteins; D004311:Double-Blind Method; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D015513:Oncogene Proteins / Q000235:genetics; D009856:Oncogene Proteins, Viral / Q000235:genetics; D027383:Papillomaviridae / Q000737:chemistry / Q000235:genetics*; D030361:Papillomavirus Infections / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011467:Prostate / Q000737:chemistry* / Q000166:cytology / Q000821:virology*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014412:Tumor Virus Infections / Q000235:genetics*
D017931:DNA Primers; D004279:DNA, Viral; D004268:DNA-Binding Proteins; C093478:E1 protein, Human papillomavirus 16; C052603:E6 protein, Human papillomavirus type 18; D015513:Oncogene Proteins; D009856:Oncogene Proteins, Viral