高危型人乳头瘤病毒阳性宫颈组织中FAM19A4甲基化检测对宫颈癌前病变的临床意义

Significance of FAM19A4 methylation detection in high-risk human papilloma virus positive cervical tissues for cervical precancerous lesions

目的探究高危型人乳头瘤病毒(HPV)阳性宫颈组织中FAM19A4甲基化检测对宫颈癌前病变的临床意义。方法选择2017年2月至2019年1月在山西省肿瘤医院就诊的115例宫颈疾病患者和65例宫颈正常的体检者。根据宫颈病变情况将受试者分为正常组65例,宫颈癌组50例,癌前病变组99例。采集三组受试者的宫颈脱落细胞进行HPV筛选,对于检出的HPV采用杂交捕获法进行DNA检测,对HPV基因型做分型检测;采用DNA提取试剂盒提取样品DNA,并采用重亚硫酸盐进行处理;通过RT-PCR和Western Blot方法检测各组样品中FAM19A4、PFAM19A4的mRNA和蛋白表达情况。结果三组受试者宫颈组织脱落细胞中HPV检出结果:正常组、癌前病变组和宫颈癌组中HPV检出率分别为7.69%(5/65)、35.35%(35/99)、48.00%(24/50),三组HPV检出率比较,宫颈癌组>癌前病变组>正常组,差异有统计学意义(P<0.05)。分子实验检测结果显示,与正常组相比,宫颈癌组和癌前病变组FAM19A4基因mRNA和蛋白表达降低,PFAM19A4基因表达升高(P<0.01);与癌前病变组相比,宫颈癌组PFAM19A4基因的mRNA和蛋白表达上调更显著(P<0.01)。结论高危型HPV阳性宫颈组织中FAM19A4甲基化程度的升高可增加宫颈癌变的概率,检测FAM19A4甲基化程度可为临床防治宫颈癌提供参考。

Objective To investigate the clinical significance of FAM19A4 methylation detection in high-risk human papilloma virus(HPV)positive cervical tissues for cervical precancerous lesions.Methods A total of 115 patients with cervical disease and 65 healthy persons who received cervical physical examination in Shanxi Cancer Hospital from February 2017 to January 2019 were selected.According to the cervical lesions,the subjects were divided into normal group(65 cases),cervical cancer group(50 cases)and precancerous lesion group(99 cases).The cervical exfoliated cells of three groups of subjects were collected for HPV screening.And for the detected HPV,DNA was detected by hybridization capture method.And the HPV genotype was tested,and DNA extraction kit was used to extract sample DNA,and the DNA was processed with bisulfite.mRNA and protein expression of FAM19A4 and PFAM19A4 in each group were detected by reverse transcription-polymerase chain reaction and western blot.Results The detection results of HPV in the cervical exfoliated cells in normal group,precancerous lesion group and cervical cancer group were 7.69%(5/65),35.35%(35/99),48.00%(24/50),respectively.In comparison on detection rate of HPV,the cervical cancer group was higher than precancerous lesion group,while precancerous lesion group was higher than normal group,and the difference was statistically significant(P<0.05).The results of molecular experiment showed that,compared with the normal group,the expression of FAM19A4 gene mRNA and protein in the cervical cancer group and precancerous lesion group were significantly lower,and the expression of PFAM19A4 gene in the two groups were significantly higher(P<0.01);and compared with the precancerous lesion group,the expression of PFAM19A4 gene mRNA and protein in cervical cancer group were significantly up-regulated(P<0.01).Conclusions The high degree of methylation of FAM19A4 in high-risk HPV-positive cervical tissue can increase the risk of cervical lesions.The detection of FAM19A4 methylation can provide reference to clinical prevention and treatment of cervical cancer.