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Abstract
This study was done at Medical Biochemistry Department, Faculty of Medicine, Ain Shams University during the period from 2006 – 2009.
The aim of this study was to investigate the impact of HPV high-risk genotypes DNA integration on HNSCC development in Egyptian patients through:
A)Direct detection of the HPV DNA in the genome of cancer tissues and comparing it with safety margin tissues from the same patient to prove that the insult of integration of the virus in the genomic DNA is linked to the process of malignant transformation as a cause-effect relationship.
B)Studying the expression of the cIAP-2 in a trial to link HPV integration with the anti-apoptotic cIAP-2 gene over-expression.
C)Investigating a possible synergistic role of the viral genome integration and other environmental factors such as smoking.
This study included 30 patients who were examined endoscopically and the tumor was resected by hot loop resection and sent for pathological staging and grading of the tumor according to (TNM) classification. Their tissue samples were stratified into two groups:
Group A: Head and neck cancer tissues.
Group B: safety margin of the cancer tissues of the same patients as our control samples.
All patients in our study were subjected to complete detailed history taking, general and local examination, and routine laboratory investigations. All patients were subjected to qualitative detection of HPV gene in their tissue samples by viral DNA extraction and detection using PCR technique.
Also, cIAP-2 gene RNA was extracted and detected by using qualitative RT PCR and semiquantitative RT PCR. In addition, RT-PCR was utilized for detection of β-actin as a house keeping genes (internal control) in all samples. The positivity rate and semi quantitative level of cIAP-2 were calculated by relative quantitation of cIAP-2 to β-actin in the same sample using, “Quantity one” computer program version 4.6.3, (Bio-Rad Laboratories, USA).
The results were calculated and statistically analyzed by the SPSS software version (15).
The results of the study showed the following:
Group I: Cancer tissue samples.
Smokers represented (66.6) % of these patients while non smokers represented (33.3%), showing a significant relation between smoking and head and neck cancer.
Qualitative PCR:The percentage of patients who were HPV positive in this group was (86.6%) declaring a high significant association between HPV positivity and head and neck cancer.
No significant difference was found between the positivity rate of HPV and any of the studied clinicopathological factors.
Qualitative and Semiquantitative RT PCR: The percentage of patients who were cIAP -2 positive in this group was (76.7%) assuming a significant relation between cIAP-2 expression and head and neck cancer.
No significant correlation was found between the positivity rate of cIAP-2 and any of the studied clinicopathological factors.
Qualitative and Semi-quantitative RT PCR assessment of cIAP-2 expression in relation to HPV positivity revealed that: cIAP-2 was expressed in (73.7%) of HPV positive patients, assuming a significant relation between cIAP-2 expression and HPV positivity.
Group II: Safety margin tissue samples.
Qualitative PCR:The percentage of patients who were HPV positive in this group was (16.7%).
Qualitative and Semi quantitative RT PCR: The percentage of patients who were cIAP -2 positive in this group was (0%).
The Overall sensitivity, specificity, positive predective values (PPV), negative predictive values (NPV) and accuracy of HPV and cIAP-2 were as follow:
*The HPV detection by Qualitative PCR was: 86.6% sensitivity, 83.3% specificity, 83.8% PPV, 86.2% NPV and 85% accuracy.
*The cIAP-2 detection by Qualitative and Semi quantitative RT PCR to differentiate cancer from safety margin tissues was:
77% sensitivity, 100% specificity, 100% PPV, 81.1% NPV and 88.3% accuracy.
*The cIAP-2 detection by Qualitative and Semi quantitative RT PCR to differentiate between HPV carcinogenicity mediated by cIAP-2 or other pathways was: 73.7% sensitivity, 100% specificity, 82.6% PPV, 100% NPV and 86.6% accuracy.