Amir Sohrabi

Detection of mucosal type human papillomavirus in cutaneous squamous cell carcinoma in Iran

Squamous cell carcinoma (SCC) is the second most common non-melanoma skin cancer. The oncogenic role of human papilloma virus (HPV) in cutaneous SCC has been suggested by several studies performed on immunosuppresed patients. However, the role of mucosal type HPV in SCC patients with normal immunity has not been studied extensively. Sixty skin biopsies from immunocompetent SCC patients and 60 benign skin specimens were evaluated for mucosal type HPV DNA using polymerase chain reaction (PCR) and immunohistochemistry (IHC). Mucosal type HPV DNA was detected in 18 of 60 cases (30%) and in 7 of 60 controls (11.6%) using PCR. HPV immunostain was positive in 16 of 60 cases (26.6%) and in 15 of 60 controls (25%). Mixed infection with HPV 18, 11, 6 was found in half of the SCC cases. The most prevalent subtype was HPV 18 followed by HPV 6 and 11. The frequency of HPV DNA was significantly elevated in our cases compared to controls (P value <0.01, OR=16.8, 95% CI: 3.3-74.9). Our findings suggest an association of mucosal type HPV, especially HPV 18, with skin SCC in Iranian patients with normal immunity.

C13orf18 and C1orf166 (MULAN) DNA Genes Methylation are Not Associated with Cervical Cancer and Precancerous Lesions of Human Papillomavirus Genotypes in Iranian Women

BACKGROUND Nowadays, molecular biomarkers have critical roles for cancer diagnosis and prognosis in clinical laboratories. Human papillomaviruses are the main agents for etiology of cervical carcinoma. The present survey was conducted to evaluate the genes methylation in cervical cancer and precancerous lesions involvement with HPV genotypes. MATERIALS AND METHODS C13orf18 and C1orf166 (MUL1 or Mulan) DNA methylation as potential biomarkers and risk factors was investigated in 112 liquid based cytology and Formalin-Fixed Paraffin- Embedded tissue specimens in Iranian females with cervical intraepithelial neoplasia and dysplasia. RESULTS In this survey, HPV18 (61.6%) and HPV16 (42.9%) proved to be the most common HPV genotypes identified by In-House Multiplex Real Time PCR. There were no significant relationship between HPV positivity and the methylated DNA genes mentioned above (p>0.05). CONCLUSIONS Our MethyLight data demonstrated that these genes could not be considered as specific, sensitive and suitable prognostic biomarkers in cervical dysplasia related HPV. It is suggested that further studies with more patients should be done on candidate methylated markers in different countries in order to plan for cervical cancer prevention.

Development of In-House Multiplex Real Time PCR for Human Papillomavirus Genotyping in Iranian Women with Cervical Cancer and Cervical Intraepithelial Neoplasia

BACKGROUND HPV related cervical cancer as one of the most common women cancers in developing countries. Regarding accessibility of commercial vaccines, any long or short term modality for integrating preventive immunization against HPV in a national program needs comprehensive information about HPV prevalence and its genotypes. The important role of selecting most accurate diagnostic technologies for obtaining relevant data is underlined by different assays proposed in the literature. The main objective of the present study was to introduce an in-house HPV typing assay using multiplex real time PCR with reliable results and affordable cost for molecular epidemiology surveys and diagnosis. MATERIALS AND METHODS 112 samples of formalin fixed paraffin embedded tissues and liquid based cytology specimens from patients with known different grades of cervical dysplasia and invasive cancer, were examined by this method and the result were verified by WHO HPV LabNet proficiency program in 2013. RESULTS HPV was detected in 105 (93.7%) out of 112 samples. The dominant types were HPV 18 (61.6%) and HPV 16 (42.9%). Among the mixed genotypes, HPV 16 and 18 in combination were seen in 12.4% of specimens. CONCLUSIONS According to acceptable performance, easy access to primers, probes and other consumables, affordable cost per test, this method can be used as a diagnostic assay in molecular laboratories and for further planning of cervical carcinoma prevention programs.

Possible Synergistic Interactions Among Multiple HPV Genotypes in Women Suffering from Genital Neoplasia

Objective: Persistence of HPV infection is the true cause of cervical disorders. It is reported that competition may exist among HPV genotypes for colonization. This survey was designed to establish the multiple HPV genotype status in our community and the probability of multiple HPV infections involvement. Methods: All multiple HPV infections were selected for investigation in women suffering from genital infections referred to private laboratories in Tehran, Iran. A total of 160 multi HPV positive specimens from cervical scraping were identified by the HPV genotyping methods, “INNO-LiPA and Geno Array”. Result: In present study, HPV 6 (LR), 16 (HR), 53 (pHR), 31 (HR) and 11 (LR) were included in 48.8% of detected infections as the most five dominant genotypes. HPV 16 was detected at the highest rate with genotypes 53, 31 and 52, while HPV 53 appeared linked with HPV 16, 51 and 56 in concurrent infections. It appears that HPV 16 and 53 may have significant tendencies to associate with each other rather than with other genotypes. Analysis of the data revealed there may be some synergistic interactions with a few particular genotypes such as “HPV 53”. Conclusion: Multiple HPV genotypes appear more likely to be linked with development of cervical abnormalities especially in patients with genital infections. Since, there are various patterns of dominant HPV genotypes in different regions of world, more investigations of this type should be performed for careHPV programs in individual countries.

Comparison of In-House Multiplex Real Time PCR, Diagcor GenoFlow HPV Array Test and INNO-LiPA HPV Genotyping Extra Assays with LCD- Array Kit for

BACKGROUND Human papillomavirus is a major etiologic agent for some human common cancers. Cervical precancer and cancer is the most prevalent dysplasia by HPV genotypes. Various rapid and sensitive methods have been developed into readily HPV genotyping. METHODS In the present study, we compared the performance of Real Time PCR, GenoFlow HPV Array, and INNO-LiPA HPV Genotyping Extra Assays with LCD- Array. RESULTS From 108 cervical samples, HPV was detected in 33 women (30.55%). Among detected HPV genotypes, HPV 6 and 11 were dominant genotypes. Comparing these methods revealed that for Real Time PCR, Genoflow, and INNO-LiPA in comparison with LCD Array, sensitivity and specificity were 94.2%, 93%; 76.7%, 93%; 64%, 96.5%, respectively. Overall, accuracy and precision of these methods were more than 80% and 90%, respectively. CONCLUSIONS It seems that these methods are reliable and suitable for detection and genotyping of HPVs in cervical disorders and other dysplasia associated with human papillomaviruses.

Deregulation of miR-21 and miR-29a in Cervical Cancer Related to HPV Infection

BACKGROUND Early diagnosis is an important factor to improve the survival of Invasive Cervical Cancer (ICC) patients. Molecular biomarkers such as micro RNA (miRNA) can be used in the early detection of ICC. The expression of miR-21 and miR-29a are deregulated in many types of human cancers. OBJECTIVE The aim of this study was to investigate the differences in miR-21 and miR-29a expression patterns in the Human Papilloma Virus (HPV) infection and various grades of cervical cancer among Iranian women. METHODS Small RNAs were extracted from positive for HPV, cervical cancer and healthy samples from 43, 50 and 46 individuals, respectively. Expression levels of miR-21 and miR-29a were analyzed by SYBR Green real-time RT-PCR using specific primers, and 5s rRNA as the internal reference gene. RESULTS Results have shown a significant increase in miR-21 and decrease in miR-29 in cancerous samples in comparison with the control groups (P < 0.0001). CONCLUSION This study illustrated that miR-21 and miR-29a could be operated as an oncogene and tumor-suppressor in cervical cancer progression. More studies are needed to demonstrate the role of miR-21 and miR-29a as potential biomarkers for the diagnosis of cervical cancer in future investigations.

Glutathione S-transferase omega gene polymorphism as biomarker for human papilloma virus and cervical cancer in iranian women

Objective: Human papillomavirus (HPV) infection is an important sexually-transmitted infection worldwide. Persistent infections with different high-risk HPV genotypes may cause cervical intraepithelial neoplasia and cervical cancer. Single nucleotide polymorphisms of glutathione S-transferase omega (GSTO) 1 and 2 play an important role in cancer progression. To evaluate GSTO gene polymorphism influence on women’s susceptibility to low-risk or high-risk HPV infections and also risk of cervical cancer development. Material and Methods: We examined 50 patients with cervical cancer, 43 patients who were positive for HPV, and 43 healthy individuals as negative controls. We used polymerase chain reaction-restriction fragment length polymorphism to determine GSTO1 A140D and GSTO2 N142D variants in study participants. Results: We found a significant association between the GSTO1 A140D gene polymorphism and HPV 6, 16, 18, 16/18 infections and cervical cancer in Iranian women. We noted a significant difference for the 140AD/142NN combination genotype between patients in the cervical cancer group and healthy controls. There were no significant differences for the GSTO2 N142D genotype and allele frequencies between the patient (i.e., cervical cancer and HPV-positive) groups and controls. Conclusion: The 140AD genotype, 140D allele, and 140AD/142NN combination genotype seem to confer a protective property in women’s susceptibility to HPV 6, 16, 18, 16/18 infections and cervical cancer. However, the GSTO2 N142D polymorphism is not associated with HPV infections and cervical cancer. It would appear that GSTO1 A140D SNPs likely play a role in the level of susceptibility to HPV-related cervical cancer.

Impact of Human Enterovirus 71 Genotypes in Meningoencephalitis in Iran

Background: Since the importance of poliovirus has diminished, as a result of its elimination in the majority of countries, non-polioviruses are emerging as causative agents of severe central nervous system (CNS) involvement. Outbreaks of enterovirus 71 (EV71)-associated CNS infections have recently been reported in Asia, Australia, and Europe. Objectives: This is the first study on genotyping of EV71 in children with meningoencephalitis to be carried out in Iran, and it was conducted in order to obtain an improved understanding of the disease burden of this virus, particularly with regard to CNS involvement. Patients and Methods: Viral RNA was extracted from 170 cerebrospinal fluid samples obtained from children aged under 8 years with a primary diagnosis of aseptic meningitis. Specific EV71 PCR was conducted to identify the genotype of the detected EV71 viruses. Results: Human enteroviruses (HEVs) were detected in 89 patients (52.3%). EV71 infection was detected in 19 (21.3%) of the 89 EV71-positive patients, and the C genotype was identified in 15 isolates. Conclusions: The C genotype should be considered as the prevalent EV71 circulating genotype in Iran, particularly in cases of aseptic meningitis.