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

Sohrabi, Amir,Mirab-Samiee, Siamak,Rahnamaye-Farzami, Marjan,Rafizadeh, Mitra,Akhavan, Setareh,Hashemi-Bahremani, Mohammad,Modarressi, Mohammad Hossein Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.16

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 C10rf166 (MULl 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

Sohrabi, Amir,Mirab-Samiee, Siamak,Modarressi, Mohammad Hossein,Izadimood, Narge,Azadmanesh, Kayhan,Rahnamaye-Farzami, Marjan Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.15

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.

Response to Letter to Editor “JAK2 V617F Mutation in Cervical Cancer Related to HPV & STIs” by Stephen E. Langabeer

Amir Sohrabi 대한암예방학회 2019 Journal of cancer prevention Vol.24 No.1

We thank you for the opportunity to reply to the Langabeer’s comments [1] on our previous manuscript [2]. The comments could be valuable for improving and clarifying some contents. Comments: Firstly, the JAK2 p.V617F (c.1849 G>T) is located in exon 12 (not exon 14) according to various literatures [3]. The acquired mutation can be presented in those patients with hematologic disorders. It appears that single nucleotide polymorphisms (SNPs) in immune mediators such as JAK2 could be associated with the risk of cervical cancer. However, these SNPs should be ascertained for justification in different communities and races with various genetic and epigenetic patterns [4]. The second argument raised by Langabeer [1] is that the RFLP method is an inefficient approach for identification of G>T transversion. During the last decades, several PCR-based assays with different analytical sensitivity and specificity parameters have been developed for clinical and diagnostic applications. These include High Resolution Melting analysis, PCR-Amplification Refractory Mutation System, Reverse Transcription-PCR, direct DNA sequencing, Real Time PCR, PCR-RFLP, Allele Specific PCR, etc. However, they have their disadvantages besides advantages. Nowadays, next generation sequencing and other high throughput approaches with high accuracy and performance are applicable in SNPs studies, although they are limited by high cost and interpretations [5]. It seems that PCR-RFLP approach can be reliable in detecting SNPs as well as other methods in developing countries. The current study has been performed in Reference Health laboratory, MOHME, Tehran, Iran. The JAK2 mutation survey is a part of our current projects in SNPs analysis of cervical cancer subjects related to sexually transmitted infections. The PCR-RFLP has been validated and verified by use of approved positive controls. In addition, all of subjects’ results have been confirmed by wild type, heterozygous and homozygous controls in each experiment. We have been trying to decrease any false positive and negative results by utilizing high quality CE and IVD marked materials and equipment. Nonetheless, the statistical analysis on these subjects would be challenging due to some missing patients’ data in that study. For improving and amending the analysis of SNPs survey, another manuscript is being prepared for better understanding of the JAK2 mutation associated with cervical malignancies as well as other potential polymorphisms in this issue. Obviously, more research is needed to achieve this endeavor for Cancer Care program.

Characterization of JAK2 V617F (1849 G > T) Mutation in Cervical Cancer Related to Human Papillomavirus and Sexually Transmitted Infections

Masoumeh Abdolmaleki,Amir Sohrabi 대한암예방학회 2018 Journal of cancer prevention Vol.23 No.2

Background: Human papillomavirus (HPV) genotypes that infect the genital tract play a main etiologic role in cervical cancer progression. Other environmental factors, such as sexually transmitted diseases and the host genetic pattern, contribute to infection persistence of the uterus and cervical epithelium in sustaining their malignancy. The Janus kinase 2 is a non-receptor tyrosine kinase in cell signaling process of tumor genesis. In the present study, JAK2 V167F mutation was distinguished in women with sexually transmitted infections, such as Herpes simplex virus 2, Chlamydia trachomatis and Mycoplasma genitalium and cervical cancer. Methods: This case-control survey was performed on 195 liquid based cytology of women specimens. Fifty, 98, and 47 samples were from women with known cervical cancer, HPV positive and HPV negative, respectively. Single nucleotide polymorphism analysis, sexually transmitted infections detection and HPV genotyping were carried out using approved PCR- RFLP, in-house multiplex TaqMan Real Time PCR and the reverse dot blot hybridization assay. Results: HPVs 6, 16, 18, 11, 31, and 51 were the most common genotypes. The prevalence rate of multiple HPV genotypes was 46.0% to 10.1%. Analysis of JAK2 V617F (1849 G > T) showed that prevalence of mutation was GG (65.1%), GA (34.9%), and TT (0%), respectively. There were no statistically significant differences between this mutation and variables of population survey (P ≥ 0.05). Conclusions: The molecular epidemiology study on the genetic polymorphisms, i.e., JAK2 V617F and other single nucleotide polymorphisms as a diagnostic tool is necessary for cancer screening and prophylactic programs.