The clinical significance of FAM19A4 methylation in high-risk HPV-positive cervical samples for the detection of cervical (pre)cancer in Chinese women

Abstract

Background: To explore the diagnostic value of FAM19A4 methylation in high-risk human papilloma virus (hrHPV)-positive cervical samples from Chinese women for estimating cervical cancer or its precancerous lesions.

Methods: Cervical samples from 215 women infected with high-risk HPV were collected by smear testing. We purposely chose 61 patients with cervical cancer, 57 with high-grade squamous intraepithelial lesions (HSIL), 31 with low-grade squamous intraepithelial lesions (LSIL), and 66 without cervical intraepithelial neoplasia (CIN) after histological confirmation. Taqman probe-based quantitative PCR (qPCR) was utilized to detect the methylation status of FAM19A4 in the cervical samples and further evaluate the use of this gene in the diagnosis of cervical cancer.

Results: (1) An increasing level of FAM19A4 methylation was detected with increasing progression of cervical lesions, with methylation rates of 10.61%(7/66), 35.48%(11/31), 56.14%(32/57) and 93.44%(57/61) in no CIN, LSIL, HSIL and cervical carcinoma samples respectively. (2) In all hrHPV-positive samples, the levels of FAM19A4 methylation in HPV16/18 groups were higher than that in 12 other hrHPV groups (P < 0.05), but there was no significant difference between two groups after grouping cervical lesions into cervical cancer, HSIL, LSIL and no CIN groups (P>0.05). (3)There were no significant differences of FAM19A4 methylation in different clinicopathological parameters of cervical cancer. (4) Though the sensitivity of FAM19A4 methylation test was inferior to that of cytology and FAM19A4 combining with HPV16/18 genotyping, but showed the best specificity with 81.44% both for detection HSIL alone and ≥ HSIL, with favorable youden index (YI) and area under curve (AUC).

Conclusion: FAM19A4 is a specific biomarker of cancerous lesions of the cervix. FAM19A4 methylation analysis may serve as an auxiliary screening method for diagnosis of cervical (pre)cancer. However, in consideration of the limitations of this retrospective study, prospective population-based studies are necessary for further confirmation of the diagnostic value of FAM19A4 methylation for detection of cervical (pre)cancer in Chinese women.

Keywords: Cervical cancer; Cytology; DNA methylation; FAM19A4; HPV genotyping; Quantitative PCR (qPCR).

Fig. 1

Flowchart of study population. 12 other hrHPV refer to HPV31, 33, 35, 39, 45 51, 52, 56, 58, 59, 66 and 68 infection

Fig. 2

Methylation scores median of different cervical lesions. In this box-and-whisker plot, the boxes represent median values; the upper and lower lines outside the boxes represent the 25th and 75th percentiles, respectively

Fig. 3

The diagnostic power of FAM19A4 methylation, cytology, HPV16/18 genotyping and the combination of FAM19A4 methylation and HPV16/18 genotyping. (a) Power of FAM19A4 methylation, cytology, HPV16/18 genotyping and the combination of FAM19A4 methylation and HPV16/18 genotyping in differentiating ≥HSIL patients from ≤LSIL patients. (b) Power of FAM19A4 methylation, cytology, HPV16/18 genotyping and the combination of FAM19A4 methylation and HPV16/18 genotyping in differentiating HSIL patients from ≤LSIL patients