HPV 16 detection in cervical lesions, physical state of viral DNA and changes in p53 gene.
BACKGROUND
Persistent infection with high risk human papillomavirus (HPV) has been linked to cervical carcinoma. Integration of viral DNA into host cell DNA is essential for this cancer development, promoting disruption of the HPV E2 gene, thus leading to unregulated increases in E6 and E7 proteins and inactivating the products of p53 and Rb tumor suppressor genes.


OBJECTIVE
To investigate HPV 16 infection in cervical lesions, physical state of viral DNA and p53 gene alterations in a group of women attending a public health service.


METHODS
Prospective, non-controlled, transversal study.


METHODS
Gynecological clinic of the School of Medicine, Universidade Federal Fluminense.


METHODS
43 consecutive patients with cervical lesions referred to our service.


METHODS
Cases were classified via cytology/histology as normal, HPV infection, condyloma, low-grade squamous intraepithelial lesion (LSIL), high-grade squamous intraepithelial lesion (HSIL) and carcinoma. HPV infection was studied via polymerase chain reaction (PCR) using two PCR primer sets, to determine DNA integration. p53 gene changes were investigated by single-strand conformation polymorphism (SSCP) analysis.


RESULTS
One normal case, 7 HPV infections, 6 condylomas, 7 LSIL, 14 HSIL and 8 cancers were found, with 95% positive for HPV genome when tested using both L1 and E6 primers. HPV 16 was most prevalent (73.1%). HPV 16 DNA was integrated within the host genome in 3 LSIL. One LSIL progressed to HSIL by 13 months after first diagnosis. Among HPV 16-positive HSIL cases, 50% contained integrated viral DNA. HPV 16 E2 gene disruption was seen in 7 cancers (87.5%). Only smal-cell carcinoma showed intact HPV 16 E2 gene. Abnormal p53 bands detected by PCR/SSCP were observed in 4 cases: 2 squamous carcinoma with parametrium (exon 8) and two cervical intraepithelial neoplasia (CIN) III (exons 5 and 7). All cases presented HPV 16 E2 gene loss.


CONCLUSIONS
The sample had a high rate of high-risk HPV detected in benign and malignant lesions; high cervical cancer burden; HPV 16 DNA integration in all except one case of cancer; p53 gene changes in CIN III and in invasive cancer cases associated with DNA integration.
do Horto dos Santos Oliveira
Ledy
L
Department of Microbiology and Parasitology, Biomedical Institute, Universidade Federal Fluminense, Niterói, Rio de Janeiro, Brazil. mipledy@centroin.com.br
Rodrigues
Eliane de Vasconcelos Machado
Ede V
de Salles Lopes
Ana Paula Terra Alvim
AP
Fernandez
André de Paula
Ade P
Cavalcanti
Silvia Maria Baeta
SM
D000293:Adolescent; D000328:Adult; D000368:Aged; D001483:Base Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000302:isolation & purification*; D005260:Female; D016158:Genes, p53 / Q000235:genetics*; D006801:Humans; D008875:Middle Aged; D009154:Mutation; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000302:isolation & purification*; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D050725:Papillomavirus E7 Proteins; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology; D016133:Polymerase Chain Reaction; D011446:Prospective Studies; D012097:Repressor Proteins; D014412:Tumor Virus Infections / Q000175:diagnosis* / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000821:virology
D017931:DNA Primers; D004279:DNA, Viral; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16