Methylation of CADM1 and MAL together with HPV status in cytological cervical specimens serves an important role in the progression of cervical intraepithelial neoplasia

Abstract

Cervical cancer (CC) is the second most common type of cancer affecting the female population. The development of CC takes several years, and involves a precancerous stage known as cervical intraepithelial neoplasia (CIN). A key factor in the development of disease is the human papillomavirus (HPV) infection, which initiates carcinogenesis. Furthermore, CC is also impacted by epigenetic changes such as DNA methylation, which causes activation or exclusion of certain genes, and the hypermethylation of cytosines in promoters, thereby switching off previously active genes. The majority of DNA methylation events occur at cytosine-guanine nucleotides, which in the human genome are known as CpG islands. The aim of the present study was to investigate the methylation levels in intronic sequences of the two tumor suppressor genes cell adhesion molecule 1 (CADM1) and T-lymphocyte maturation associated protein (MAL) using cytological samples and to identify potential biomarkers involved in CIN by pyrosequencing. DNA was isolated from cervical smears from patients with CINs, with healthy patients serving as a control group. Samples were converted by treatment with sodium bisulfite and subsequent pyrosequencing to detect the methylation status of the selected genes. The presence of HPV DNA infection analyzed by the polymerase chain reaction, was detected in each sample. Of the total number of samples (n=91), the present study confirmed the presence of one or two high-risk subtypes of HPV in 39 cases (42.85%) and HPV infection was significantly associated with CIN2+ lesions. For the two genes (MAL and CADM1) the present study confirmed that the median methylation was significantly higher in HPV positive patients [P=0.0097, 95% confidence interval (CI): (−0.030, −0.003)/P=0.0024, 95% CI: (−0.06, −0.01)] when compared with patients negative for HPV DNA infection, and the average methylation was demonstrated to be increased with the degree of cervical lesion. The present study used logistical regression to model the dependence between the case/control statuses (control group vs. Dg. 1–4). The area under the curve values for MAL were: 84% for cervical inflammation, 71% for CIN1, 73.4% for CIN2+ and 77% for squamous cell carcinoma (SCC); and for CADM1 were: 88.6% for cervical inflammation, 68% for CIN1, 80% for CIN2+ and 89% for SCC. The present study confirmed that there were statistically significant differences between the methylation levels of individual CpGs and significantly higher median methylation in patients positive for HPV16/18. CADM1 exhibited higher levels of methylation in almost every study group when compared with MAL during the transition of CIN and appeared to be a promising biomarker for future study.

Plasma CADM1 promoter hypermethylation and D‐dimer as novel metastasis predictors of cervical cancer

“…It is the same as to cervical cancer. Downregulated CADM1, caused by its promoter hypermethylation, was involved in the metastasis and prognosis in CC patients . However, to the best of our knowledge rare studies have investigated CADM1 promoter hypermethylation from blood samples.…”

“…Downregulated CADM1, caused by its promoter hypermethylation, was involved in the metastasis and prognosis in CC patients. [28][29][30] However, to the best of our knowledge rare studies have investigated CADM1 promoter hypermethylation from blood samples. Therefore, we firstly showed the high coincidence rate of CADM1 promoter methylation in pair-matched tissue and plasma samples of CC patients.…”

“…Written informed consent for the use of the clinical samples was obtained from all patients and controls. All the women had the following median ages: 40 years (range [30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45]…”

DNA Methylation and Hydroxymethylation in Cervical Cancer: Diagnosis, Prognosis and Treatment

“…It is important that the sensitivity, specificity, and rate of methylation of all the biomarkers are similar ( Table 2). Mersakova et al (2018) and Rong et al (2019) have recently shown that CADM1 is a potential biomarker for cervical cancer. There was a significant difference in the promoter Kan et al, 2014;Lai et al, 2014;Kong et al, 2015;Nikolaidis et al, 2015;Xu et al, 2015Xu et al, , 2018Chen et al, 2016;Liou et al, 2016;Huang et al, 2017;Fang et al, Lee et al, 2006;Yeon et al, 2018 methylation of plasma CADM1 and its D-dimer between healthy individuals and those with cervical cancer.…”

Frequent promoter methylation of HOXD10 in endometrial carcinoma and its pathological significance

“…Therefore, DNA methylation may be used as a marker in early cancer screening (24,(42)(43)(44). Hypermethylation of specific TSG, as well as low mRNA and protein expression levels, has been reported in a number of human malignancies, including EC (28,45,46). HOXD10 is one of the ANTP homeobox genes located on chromosome 2 (2p31).…”