Cytological DNA methylation for cervical cancer screening: a validation set

Abstract

BackgroundIn a previous training set with a case-controlled design, cutoff values for host EPB41L3 and JAM3 gene methylation were obtained for the detection of cervical intraepithelial neoplasia (CIN) 2 or more severe lesions (CIN2+). This validation trial was conducted to evaluate the role of DNA methylation in screening for CIN2+ by cervical cytology among unselected participants.MethodsFrom June 1, 2019, to September 1, 2019, in our study center, we collected liquid-based samples from cervical swabs for methylation assays and hrHPV testing in eligible patients. The primary endpoint was the diagnostic accuracy of DNA methylation and hrHPV genotyping for CIN2+ according to confirmed histology results.ResultsAmong 307 participants, compared with hrHPV testing, the methylation assay for CIN2+ had lower sensitivity (68.7% versus 86.1%, p=0.002) but higher specificity (96.7% versus 0.696, p<0.001). The methylation assay also had favorable sensitivity and specificity in patients with negative hrHPV testing (56.3% and 96.9%) and in patients with cervical adenocarcinoma (73.7% and 92.7%). DNA methylation had higher specificity than the hrHPV assay (100.0% versus 44.4%, p<0.001) for identifying residual CIN2+ in patients without residual lesions. Positive cervical DNA methylation was associated with a diagnostic probability of endometrial carcinoma (odds ratio 15.5 [95% confidence interval 4.1-58.6]) but not of ovarian epithelial carcinoma (1.4 [0.3-6.5]).ConclusionsThe host EPB41L3 and JAM3 gene methylation assay in cervical cytology had favorable diagnostic accuracy for CIN2+ and was highly specific for residual CIN2+ lesions The methylation assay is a promising triage tool in hrHPV+ women, or even an independent tool for cervical cancer screening. The methylation status in cervical cytology could also serve as a prognostic biomarker. Its role in detecting endometrial carcinomas is worthy of further exploration.

The research progress on radiation resistance of cervical cancer

“…The combination of high-risk HPV positive testing aids in the diagnosis of CIN2+ and the evaluation of prognosis ( 91 , 92 ). The methylation of EPB41L3 and JAM3 remains unaffected by high-risk HPV and demonstrates good diagnostic accuracy for CIN2+ ( 93 ). Furthermore, previous study has indicated that the detection of gene methylation (PHACTR3, PRDM14, GHSR, FAM19A4, SST, and ZIC1) in urine is beneficial for diagnosing cervical cancer ( 94 ).…”

Diagnostic Accuracy of DNA-Methylation in Detection of Cervical Dysplasia: Findings from a Population-Based Screening Program

“…The potential applicability of the methylation test in combination with other highly sensitive methods (e.g., the HPV-DNA test) should be considered to improve the sensitivity of the test in detecting grade ≥ 1 colposcopy lesions. In addition, further risk stratification could be achieved considering HPV-genotype, which has been suggested as a triage for HPV-positive women [ 27 , 31 ]. Our results confirmed the higher accuracy of the methylation assay when combined with cytology and HPV testing.…”

“…The cytological examination was based on TBS-2001 criteria [11]. The HPV genotyping was carried out after nucleic acid extraction by QIAamp DNA Mini Kit (Qiagen, Hilden, Germany) [14], using the Anyplex II HPV HR detection kit (Seegene Inc., Seoul, Republic of Korea), allowing the detection of 14 HR-HPV genotypes, including HPV- 16,18,31,33,35,39,45,51,52,56,58,59, 66, and 68, as previously described [15,16].…”