FAM19A4基因启动子甲基化在高危型HPV阴性宫颈癌诊断中的作用

The role of FAM19A4 gene promoter methylation detection in the diagnosis of high-risk HPV-negative cervical cancer

目的探讨FAM19A4基因启动子甲基化在高危型HPV(hr-HPV)阴性宫颈癌诊断中的作用。方法收集2013年6月至2017年12月在广东省妇幼保健院妇科治疗的宫颈癌患者共490例,其中术前宫颈刷液荧光定量PCR检测HPV结果为hr-HPV阴性的患者共25例,提取石蜡包埋标本中的DNA,依次经荧光定量PCR、HC-II、高通量靶向测序3种检测方式检测HPV型别,并评估hr-HPV阴性宫颈癌的临床特征,进一步检测标本中FAM19A4基因甲基化的表达情况。结果 25例样本中经荧光定量PCR及HC-II方法检测结果为hr-HPV阴性共12例(48.0%,12/25),将12例样本进行高通量测序,仍为阴性有9例(36%,9/25);hr-HPV阴性宫颈癌标本的FAM19A4基因甲基化检出率为100%(9/9);hr-HPV阴性宫颈癌与hr-HPV阳性宫颈癌相比,在组织病理学有统计学差异(P <0.05)。结论 hr-HPV阴性宫颈癌一种临床病理较为特殊的宫颈癌,经过更加敏感的检测方法检测后,发病率将进一步下降,结合FAM19A4基因启动子甲基化检测将对hr-HPV阴性宫颈癌的诊断有重要的辅助意义。

Objective To investigate the role methylation of FAM19A4 gene promoter methylation in tissue the diagnosis of high risk HPV(hr-HPV) negative cervical cancer. Methods A total of 490 patients with cervical cancer who were treated by gynecology in Guangdong Provincial Maternal and Child Health Hospital from June 2013 to December 2017 were enrolled. Quantitative PCR was used to detect HPV using preoperative cervical brushing liquid and 25 cases of hr-HPV negative were found. Then the DNA extracted from FFPE tissues of the 25 hr-HPV negative were used to confirm the HPV existence by quantitative PCR,HC-II and High throughput sequencing. The clinical characteristics of hr-HPV negative cervical cancer were evaluated,and the status of FAM19A4 gene methylation in the specimen was detected by qMS-PCR. Results (1) The FFPE DNA results of using quantitative qPCR and HC-II were the same. In the 25 samples,hr HPV was negative in 12 cases( 48.0%, 12/25).12 samples were DNA sequenced and 9 sample were still negative( 9%,36%,9/25).(2) The methylation of FAM19A4 gene methylation was 100%(9/9) in the hr HPV negative cervical cancer group.(3) Compared with hr HPV positive cervical cancer,hr HPV negative cervical cancer was statistically different in histopathology and clinical stage(P<0.05). Conclusions hr HPV negative cervical cancer is a kind of cervical cancer with special clinicopathological features. After more sensitive detection,the incidence will be further reduced. Combined with the FAM19A4 gene promoter methylation assay will have important auxiliary significance for the diagnosis of hr-HPV negative cervical cancer.