|Year : 2021 | Volume
| Issue : 1 | Page : 55-58
Detection of human papilloma virus in prostate tissue from prostatic cancer patients in AL Najaf, Iraq
Anwar Abd Hashim Al Ramahy
Department of Medical Microbiology, Al-Sadr Medical City Hospital, Najaf, Iraq
|Date of Submission||04-Nov-2020|
|Date of Acceptance||24-Dec-2020|
|Date of Web Publication||13-Mar-2021|
Dr. Anwar Abd Hashim Al Ramahy
Department of Medical Microbiology, Al-Sadr Medical City Hospital, Baghdad
Source of Support: None, Conflict of Interest: None
Background: The prostatic tumor was the third form of cancer in males being >55 years of age, after the pulmonary and colon cancer. The prostate was infected by many viruses causing repeated or chronic inflammation or enhance carcinogenesis. The aim of this study was to determine the presence and prevalence of human papillomavirus (HPV) in the prostate tissue of patients with prostate cancer. Methods: Prostate tissue samples were collected from paraffin embedding block of 70 patients diagnosed with HPV from archives of AL-Sadder Hospital in AL-Najaf City and some archives of private histopathology laboratories from January 2016 to December of the same year. We investigated the presence of HPV DNA using the polymers chain reaction technique polymerase chain reaction (PCR). Results: The sample size was 60 patients and 10 as control. The number of benign patients is 40/60 (66.7%), whereas the number of malignancies patients is 20/60 (33.3%). There are 13/60 prostatic patients who have HPV. Eight patients of them have a malignancy prostatic tumor and five with a benign prostatic tumor. There is a highly significant association between HPV and malignancy compared with the benign prostatic disease (P = 0.016). PCR results were showed a highly significant specificity (P < 0.001) to detect the presence of HPV in malignancy and benign prostatic disease. PCR results showed a highly significant specificity (P < 0.001) to detect the presence of HPV in malignancy and benign prostatic disease. Conclusions: Our study showed an association between prostatic disease and the presence of HPV DNA in samples, but there is a highly significant difference in HPV prevalence between the malignant prostate cancer and benign prostate tissue samples.
Keywords: Benign prostatic hyperplasia, human papillomavirus, polymerase chain reaction, prostate cancer
|How to cite this article:|
Al Ramahy AA. Detection of human papilloma virus in prostate tissue from prostatic cancer patients in AL Najaf, Iraq. Biomed Biotechnol Res J 2021;5:55-8
|How to cite this URL:|
Al Ramahy AA. Detection of human papilloma virus in prostate tissue from prostatic cancer patients in AL Najaf, Iraq. Biomed Biotechnol Res J [serial online] 2021 [cited 2022 May 19];5:55-8. Available from: https://www.bmbtrj.org/text.asp?2021/5/1/55/311093
| Introduction|| |
Prostate cancer is one of the life-threatening disorders and the most common malignancy seen worldwide in men with >160 000 new cases each year in the U.S.A. It was also considered the most common cancer in males in 84 countries.
Viral infections, in particular, may lead to chronic inflammation of the prostate and initiation or development of prostate cancer. The prostate tissue is prone to sexually transmitted infection with several viruses that having oncogenic potentials. The oncogenic virus includes polyomaviruses (SV40), human papillomavirus (HPVs), and members of the herpes virus family.
Human papillomaviruses (HPV) are a small group of nonenveloped viruses with circular double-stranded DNA genomes with sizes close to 8 kb belonging to the papillomaviridae family with strong similarities to polyomaviruses. Papillomaviruses infect epithelial cells and depend on epithelial differentiation for completion of their life cycle. The expression of viral gene products was closely regulated as the infected basal cell migrates toward the epithelial surface. Expression of E6 and E7 in the lower epithelial layers drives cells into synthetic phase (S-phase), which creates an environment that was conducive for viral genome replication and cell proliferation, genome amplification, which is necessary for the production of infectious virions, was prevented until the levels of viral replication proteins rise, and depends on the co-expression of several viral proteins.
HPV is the most prevalent sexually transmitted infection in the USA. Over of the 100 HPV genotypes, approximately 60 were considered pathogenic for the genital tract and were classified by oncogenic potential. High-risk genotypes, including HPV16, 18, and 33, were more commonly associated with cancers, whereas low-risk types such as HPV6 and 11 typically cause genital warts., HPV is associated with prostate cancer disease. The association between HPV infection and cervical neoplasm was established after the link between genital HPV infections and cervical cancer was first demonstrated in the early 1980s by de Villiers EM. The involvement of HPV infection in a malignant transformation was first discovered by Hausen Zur, who identified a subgroup of HPVs as etiological agents of cervical cancer., A recent meta-analysis study showed a significantly increased risk of prostate cancer with the positivity of overall HPV detected in prostate tissues.
The prevalence of HPV DNA in tumor tissue of patients with prostate cancer has been poorly investigated in Iraq. Therefore, the present study examined the prevalence of high- and low-risk HPV, using a polymerase chain reaction (PCR) technique in AL-Najaf/Iraq.
The study protocol was approved by the Ethical Committee University of Kufa Faculty of Science on September 17, 2017, approval number 3723.
Paraffin embedding blocks were collected for 70 patients who were diagnosed with prostate cancer (20 malignant prostates, 40 benign, and 10 controls) from archives of AL-Sadder Hospital in AL-Najaf City and some archives of private histopathology laboratories from January 2016 to December of the same year.
Prostate cancer was confirmed by a pathologist through standard criteria. Tumors were staged using standard criteria by the Gleason score. Demographic and medical information including age, habitat, and Gleason score was collected from patients' medical records.
The use of paraffin-embedded prostate cancer tissues and the related clinical information were approved by the Research Ethical Committee in the AL-Najaf Health Directorate/Iraq.
The DNA extraction was done according to the commercial kit (Geneaid Biotech Ltd.). To extract DNA, up to 25 mg of tissue cut into 10 Mm fragments was incubated in 200 μl of GST and 20 μl of proteinase K than at 60°C overnight or until complete fragment digestion. Subsequently, the cell lysis was done by adding 200 ml of GSB buffer and shake vigorously for 10 s, and the total product was stored at −20°C freezer to use for the detection of HPV.
Detection of L1 gene HPV
The nucleotide sequences of the forward and reverse primer used for PCR are 5-TGG-ACA-ACA-CAA-GAA-ACA-CAA-ACT-3 and 5-ACC-GGA-GCA-TAT-ACG-GAT-CTT-CCT-3, respectively. (Gen Bank: AJ000151.1). PCR master mix was prepared using (AccuPower PCR PreMix Kit) included 1.5 μl of each specific primer, 12 μl of PCR water, and 5 μl of DNA then placed in standard AccuPower PCR PreMix Kit that containing all other components which needed to PCR reaction such as Taq DNA polymerase, dNTPs, Tris-HCl pH: 9.0, KCl, MgCl2, stabilizer, and tracking dye.
The amplification was reformed by including the reaction mix for 30 cycles in a thermocycler. Each cycle consisted of denaturation of DNA at 95°C for 30 s, followed by annealing at 56°C for 30 s, and extension at 72°C for 1 min with initial delay for 3 min at 95°C at the beginning of the first cycle and 5 min delay at 72°C at the end of the last cycle.
The reaction products were separated on 1% agarose gels with ethidium bromide at 100 V for 60 min. PCR products 342 bp were visualized using a UV transilluminator, as shown in [Figure 1].
|Figure 1: Agarose gel electrophoresis image appear the polymerase chain reaction product analysis of HPV, from in DNA extracted from FFPE prostate tissue section samples, where ladder 2000-100 bp, some positive DNA samples for HPV at 342 bp polymerase chain reaction product size|
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Statistical analysis was done using Statistical Package for the Social Sciences (SPSS) version 21 (GraphPad Software, San Diego, California, USA). Values of P < 0.05 and P < 0.001 were measured to be statistically significant.
| Results|| |
[Table 1] shows the characteristics of patients with prostatic disease, whereas the distribution of prostate cancer according to the presence of HPV by using PCR is shown in [Table 2].
|Table 2: Distribution of prostate cancer according to the presence of human papillomavirus using polymer chain reaction|
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It was observed that the age range of patients was between 50 and 90 years. The resident of patients in urban was more than in rural (66.67% of prostate patients in urban). The sample size was 60 patients and 10 as control. The number of benign patients is 40/60 (66.7%), whereas the number of malignancies patients is 20/60 (33.3%). There are five types of prostatic patients according to the Gleason grade that are used for differentiating prostatic cell carcinoma. The total number of malignant prostate was 20 and the distribution is as the following: Grade 1 was 6/20 (30%), Grade 2, 3, and 5 was 4/20 (20%), and Grade 4 was 2/20 (10%). The immunohistochemistry (IHC) test was positive in 13 patients from 20 malignancy patients, whereas only seven patients from 40 benign patients had positive results in IHC.
There are 13/60 prostatic patients who have human papillomavirus. Eight patients of them have a malignancy prostatic tumor and five with a benign prostatic tumor. On the other hand, there are 47/60 prostatic patients without HPV. There are twelve patients with malignancy and only 35 patients with benign prostatic patients. There is a highly significant association of HPV with malignancy prostatic disease compared with benign (P = 0.016).
PCR results showed a highly significant specificity (P < 0.001) to detect the presence of HPV in malignancy and benign prostatic disease.
| Discussion|| |
Prostate cancer was the second cancer type which leads to death for men. Age in this study ranges between 50 and 90 years. Many studies agreed with this study. For example, a study by Kerman's Southeast of Iran showed that the life stage 65–74 years is the highest stage of prostatic carcinoma. Saudi Arabia showed that most patients with prostatic carcinoma are between 70 and 79 years. Another study showed that the significant age of the patients with prostatic carcinoma was between 70 and 79 years. The variation in life stages between young and old men and decrease of this percentage in this stage of life may be due to the fact that the youngest men did not examine at the beginning of infection until they get older and most of the life stage 81–90 was a death in prostatic carcinoma before reaching this life stage.
This finding is in agreement with a study conducted in Kerman, south of Iran, which appears over 60% of the total percentage for low Grade 2–4 while a low percentage (about 16%) in high grade. Another study showed that the high percentage of prostatic carcinoma in Grade 5–7 was 37/59 (62%), while in low percentage in Grade 2–4 was 14/59 (23.72%). Further, it showed that the highest percentage of prostatic carcinoma in Grade 5–7 was 41/74 (57.7%), and a low percentage of prostate carcinoma in Grade 2–4 was 4/74 (5.6%). This variance in percentage was due to genetic, racial environmental factors, and dietary factors.
The present study appeared statistically high risk of prostatic cancer in Urban. This study agrees with Kerman South East of Iran which showed a high percentage in Urban (75%) and 25% in Rural this in prostatic cancer, while in beige prostate hyperplasia it was (96%) in Urban and (4%) in Rural. Another study showed that Jamaican men with prostate cancer were with a high percentage in rural (75%) compared with urban (25%). Another study showed that the percentage of prostatic carcinoma was higher in rural in (86%) than in urban. This variance has been due to the difference between urban and rural; the lack of information of patients in rural areas could get or miss the panel diagnosis for prostatic diagnosis.
This study appeared that HPV was found in both malignant and benign prostate tumors. Further, it was found that the percentage of HPV is higher in malignancy prostate tumors. This study agreed with previous studies that showed that the HPV was found in both malignant and benign prostate cancer as 70% in malignant and 20% in benign. Other studies showed that high HPV was found in both malignant and benign prostate tumors. The study showed the percentage of HPV in prostate cancer was 7/10 (70%), whereas in benign prostate was 2/10 for HPV.
The mechanism of HPV infections and prostate cancer development is far from clear. It has been proposed that exposure to environmental factors such as dietary carcinogens, infectious agents, and hormonal imbalances leads to injury of the prostate gland.
In the previous meta-analysis, the studies that used PCR-based methods to detect HPV DNA demonstrated a higher sensitivity compared with non-PCR-based methods. The variations in the sensitivity of HPV detection may be due to the differences in amplification efficiency between various types of HPV primers. Therefore, it is possible that the detection rates of HPV using HPV type-specific PCR primers may be higher compared with those using other PCR primers.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Table 1], [Table 2]