Human papillomavirus infection among Egyptian females with cervical carcinoma: relationship to spontaneous apoptosis and TNF-alpha.
OBJECTIVE
This study was designed to detect HPV type-16 in Cervical carcinoma (CC) tissue specimens. The results were correlated with clinicopathological parameters of the carcinoma, with spontaneous apoptosis and with immunoreactivity to TNF-alpha antibodies.


METHODS
Fresh frozen tissue specimens representing 30 cases of cervical carcinoma as well as 20 normal cervical tissues (NCT) were the subjects of this study. HPV-16 DNA was detected by Polymerase Chain Reaction (PCR). The occurrence of spontaneous apoptotic cell death was analyzed by the apoptosis assay. Apoptotic cells were also counted by light microscopy and the apoptotic index (AI) was calculated. Electron microscopy was used to confirm the morphology of apoptotic cells. TNF-alpha was quantified using EIA kit.


RESULTS
HPV-16 DNA was more frequent in CC than in NCT. No correlation was observed between HPV infection and grade, stage or pathologic type of CC. The occurrence of spontaneous apoptosis was significantly higher in CC than in NCT, where it was correlated to advanced tumor stage and tumor pathology being more in adenocarcinoma (AC) than in squamous cell carcinoma (SCC). Moreover, AI was negatively correlated to HPV-16 infection. TNF-alpha levels were significantly higher in CC vs. NCT, where they were positively correlated to advanced tumor stage. TNF-alpha levels were correlated to DNA fragmentation and AI (r = 0.47 and 0.57 respectively). A cut-off value for TNF-alpha was calculated to be 9.1 pg/mg protein (using ROC curve). At the determined cut-off point the sensitivity was 70% and the specificity was 80%.


CONCLUSIONS
HPV infection, high levels of TNF-alpha and spontaneous apoptosis were strongly associated with malignant phenotype of cervical tissues. Rate of spontaneous apoptosis was higher in AC compared to SCC. On the other hand, HPV negativity was correlated with AI. Moreover, TNF-alpha and apoptotic cell death were correlated to each other as well as to tumor progression. No correlation was detected between TNF-alpha and HPV-16 infection.
Ahmed
M I
MI
Oncology Diagnostic Unit, Biochemistry Dept., Ain Shams Faculty of Medicine, Abbassia, Cairo, Egypt 11566. mahaim@hotmail.com
Salahy
E E
EE
Fayed
S T
ST
El-Hefnawy
N G
NG
Khalifa
A
A
D000230:Adenocarcinoma / Q000150:complications / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D000328:Adult; D000368:Aged; D017209:Apoptosis; D002294:Carcinoma, Squamous Cell / Q000150:complications / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D002584:Cervix Uteri / Q000166:cytology / Q000821:virology; D015897:Comorbidity; D004279:DNA, Viral / Q000302:isolation & purification; D004534:Egypt; D005260:Female; D006801:Humans; D008875:Middle Aged; D009367:Neoplasm Staging; D027383:Papillomaviridae / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000150:complications / Q000175:diagnosis* / Q000821:virology; D016133:Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis* / Q000378:metabolism; D014412:Tumor Virus Infections / Q000150:complications / Q000175:diagnosis* / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000150:complications / Q000378:metabolism / Q000473:pathology / Q000821:virology*
D004279:DNA, Viral; D014409:Tumor Necrosis Factor-alpha