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 (cid:83)(cid:82)(cid:87)(cid:72)(cid:81)(cid:87)(cid:76)(cid:68)(cid:79)(cid:3)(cid:69)(cid:76)(cid:82)(cid:80)(cid:68)(cid:85)(cid:78)(cid:72)(cid:85)(cid:86)(cid:3)(cid:68)(cid:81)(cid:71)(cid:3)(cid:85)(cid:76)(cid:86)(cid:78)(cid:3)(cid:73)(cid:68)(cid:70)(cid:87)(cid:82)(cid:85)(cid:86)(cid:3)(cid:90)(cid:68)(cid:86)(cid:3)(cid:76)(cid:81)(cid:89)(cid:72)(cid:86)(cid:87)(cid:76)(cid:74)(cid:68)(cid:87)(cid:72)(cid:71)(cid:3)(cid:76)(cid:81)(cid:3)(cid:20)(cid:20)(cid:21)(cid:3)(cid:79)(cid:76)(cid:84)(cid:88)(cid:76)(cid:71)(cid:3)(cid:69)(cid:68)(cid:86)(cid:72)(cid:71)(cid:3)(cid:70)(cid:92)(cid:87)(cid:82)(cid:79)(cid:82)(cid:74)(cid:92)(cid:3)(cid:68)(cid:81)(cid:71)(cid:3)(cid:41)(cid:82)(cid:85)(cid:80)(cid:68)(cid:79)(cid:76)(cid:81)(cid:16)(cid:41)(cid:76)(cid:91)(cid:72)(cid:71)(cid:3)(cid:51)(cid:68)(cid:85)(cid:68)(cid:73)(cid:192)(cid:81)(cid:16)


Introduction
Cervical cancer associated with Human papilomaviruses especially with High Risk (HR) HPV genotypes is one the most public health problems in developing countries such as Iran.Based on the data reported by WHO, the crude incidence and mortality rates of HPV related cervical cancer until 2010 in Iran are about 1.8 and 0.8 per 100,000 per year, respectively.HPVs 16(HR), 18(HR), 31(HR), 6(LR), 11(LR), 33(HR) are the most common HPV genotypes in Iranian women with and without cervical dysplasia (WHO 2010;Laudadido 2013;Shayanfar et al., 2013).It is proposed that individual genetic and epigenetic characteristics accompanied with HPV infection in cervical cancer have important role as main risk factors in cancer progression (Szalmás and Kónya., 2009;Wentzensen et al., 2009;Mohiuddin et al., 2013;Wang ., 2014).Over the last decade, evaluation of methylation biomarkers in cervical precancerous and cancer have been done in various studies.Although wide attempt have been lesions admitted to Tehran Women Central hospital, Imam Khomeini Hospital and for each patient and clinical data were entered.The ethical considerations of the study were approved and a consent from was signed by each patient.Then Liquid Based Cytology (LBC) preservatives and FFPE blocks were transported to molecular biology department in health reference laboratory of ministry of health and medical

Statistical analysis
of C13orf18 and c1orf166 genes and existence of Human Papillomaviruses, an using SPSS software version 18.5.

Discussion
Cervical carcinoma as a social problem with high morbidity and mortality in the worldwide is discussed.Insomuch, human papillomaviruses genotypes are considered as the main cause of malignancies particulary cervical cancer.Epigenetic and polymorphisms patterns as diagnostic biomarker and HPV genotypes detection can be used for determining appropriate treatment strategy for early stages of cervical cancer and increase survival in cervical malignancies (Nunobiki et al., 2011;Hwang and Shroyer 2012;Wang 2014).Therefore, the extensive studies have been done on the methylation analysis of various genes in different societies (Yang et al., 2009;Eijsink et al., 2011;2012;Snellenberg et al., 2012;Sun et al., 2012;Banzai et al., 2013;Lan et al., 2013;Al-Shabanah et al., 2014;Ma et al., 2014).Functional and MethyLight analysis of these genes shown that C13ORF18 gene might represents cervical cancer suppressive activities upon re-expression and C1ORF166, now known ligase that regulates the organelle's dynamics and NF-kB signaling.It is possible that these genes have functional role during cervical tumor development and progression (Li et al., 2008;Yang et al., 2009;Huisman et al., 2013).The distribution of HPV 16 and HPV 18 along with other high risk HPV genotypes in this study is similar to other surveys (Clifford et al., 2005;Lee et al., 2011;Kasamatsu et al., 2012;Manjari and Sweta., 2012;Hamzi Abdul Raub et al., 2013;Turki et al., 2013;Wang et al., 2013;Othman and Othman., 2014).Although the case studies have been conducted on methylation status of C13ORF18 and C1orf166 genes (Yang et al., 2009;Eijsink et al., 2011;Eijsink et al., 2012).Likely, the methylated genes could be considered as proper diagnostic biomarkers in cervical cancer mid HPVs (Dueñas-González et al., 2005;Szalmás et al., 2009;Wentzensen et al., 2009;Nogueira da Costa and Herceg., 2012).However, they were detected of DNA methylated genes for C13orf18; 65 (58%) and C1orf166; 45 (40%) in this research, but we did not found dysplasia and HPV genotypes previously found positive (p>0.05).Therefore, introducing these methylated genes as prognostic biomarkers of cervical cancer needs further studies with genetic patterns variation and various designs and acceptable number of clinical specimens.
In Conclusions, Obviously increased cervical cancer and precancerous lesions with HPV genotypes infection were seen in different regions of world, hence, further investigations should be emphasized for detection and development of molecular diagnostic biomarkers in cancers.More investigations on CpG methylation regions in different countries with more clinical samples biomarkers and treatment procedures in cancers.

Figure 1 .
Figure 1.Frequency Rate of C13orf18 and C1orf166 DNA Genes Methylation in Cervical Disorders with cervical disorders.The details of prevalence of HPV genotypes and cervical disorder and Cervical Intraepithelial Neoplasia, and frequency rate of DNA genes methyaltion were summarized in Table1and Figure1.