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. 2022 Jan 1;23(1):143-149.
doi: 10.31557/APJCP.2022.23.1.143.

Detection of Global DNA Methylation in Cervical Intraepithelial Neoplasia and Cancerous Lesions by Pyrosequencing and Enzyme-Linked Immunosorbent Assays

Rungtip Thumbovorn  1   2 , Parvapan Bhattarakosol  1   3 , Arkom Chaiwongkot  1   3
Affiliations

Detection of Global DNA Methylation in Cervical Intraepithelial Neoplasia and Cancerous Lesions by Pyrosequencing and Enzyme-Linked Immunosorbent Assays

Rungtip Thumbovorn et al. Asian Pac J Cancer Prev. .

Abstract

Background: Cervical cancer is one of the most significant cancer found in women worldwide especially in developing countries. Previous reports showed that global DNA hypomethylation was correlated with various types of cancer including cervical cancer.

Methods: Long interspersed nuclear element-1 (LINE1) pyrosequencing and Enzyme linked-immunosorbent assay (ELISA) assays were used for detection of global DNA methylation. The ELISA results were compared to bisulfite LINE1 pyrosequencing assay.

Results: Different cervical cancer cell lines (CaSki, SiHa, HeLa, ME180, MS751, C33A) showed low global methylation percentage when compared to normal white blood cells by ELISA assay (1.47%-5.09% vs 8.20%, respectively) and by LINE1 pyrosequencing (20%-45% vs 62%, respectively). Global DNA methylation levels in cervical cancer samples were lower than precancerous lesions (Normal-CIN3) by LINE1 pyrosequencing (mean, 48.8% vs 56.9%, respectively, p<0.05) and ELISA assay (mean, 3.03% vs 3.85%, respectively, p<0.05).

Conclusion: Global DNA hypomethylation was predominantly found in cervical cancer samples detected by ELISA and LINE1 pyrosequencing assays and could be used as triage tests in cervical cancer screening. ELISA assay is a suitable method for detection of global DNA methylation in large population; however, it should be further evaluated in a large clinical samples in order to be used as screening method.

Keywords: Cervical cancer; Long interspersed nuclear element-1 (LINE1); Pyrosequencing; enzyme linked immunosorbent assay; global DNA methylation.

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Conflict of interest statement

The authors declare that there is no conflict of interest.

Figures

Figure 1
Figure 1
Methylation Levels in 167 Cervical Samples with Different Lesions Severity (*=p<0.05 (Kruskal wallis))
Figure 2
Figure 2
Methylation Levels in Ninety-Seven Cervical Samples with Different Lesions Severity Detected by ELISA assay (*=p<0.05 (Kruskal wallis))
Figure 3
Figure 3
Correlation of Bisulfite LINE1 Pyrosequencing and ELISA Assays (N=58)
Figure 4
Figure 4
Receiver Operating Characteristic (ROC) Curve Shows the Sensitivity and Specificity of Methylation Levels to Differentiate between Precancerous (Normal,CIN1,CIN2-3) and Cancerous Lesions by Bisulfite LINE1 Pyrosequencing
Figure 5
Figure 5
Receiver Operating Characteristic (ROC) Curve Shows the Sensitivity and Specificity of Methylation Levels to Differentiate between Precancerous (Normal,CIN1,CIN2-3) and Cancerous Lesions by ELISA Assay
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