PAX1检测对宫颈上皮内瘤变和宫颈癌HPV感染的临床诊断价值

摘要

目的 探讨配对盒家族基因1(paired boxed gene 1,PA X 1)的甲基化定量检测以及免疫组化法等对高级别宫颈癌前病变的临床诊断价值.方法 选取2014年1月至2016年10月在上海交通大学附属第六人民医院就诊的135例高危亚型人乳头瘤病毒(human papilloma virus,HPV)感染者为研究对象,根据病理诊断结果将其分为正常宫颈组(组Ⅰ,40例)、低级别宫颈癌前病变组[宫颈上皮内瘤变(cervical intraepi-thelial neoplasia,CIN)Ⅰ,组Ⅱ,44例]及高级别宫颈癌前病变组(CINⅡ~Ⅲ,组Ⅲ,51例).采集各组患者宫颈组织标本,分别采用免疫组织化学法、免疫印记法、聚合酶链反应(polymerase chain reaction,PCR)、PA X 1定量甲基化聚合酶链反应及焦磷酸测序法检测各组样本中的PAX1甲基化水平及表达情况,绘制患者PAX1甲基化水平接受者操作特征曲线(receiver operating characteristic curve,ROC曲线).结果 组Ⅰ的PA X 1甲基化水平为(4.53±2.62)%,组Ⅱ为(3.24±2.28)%,组Ⅲ为(13.67±5.38)%,三组比较差异具有显著性(F=22.35,P<0.001).ROC曲线下面积为0.9216.PA X 1甲基化水平对高级别宫颈癌前病变的诊断阈值为8.74%,灵敏度为86.27%,特异度为98.81%.结论 采用焦磷酸测序法定量检测宫颈组织中PA X 1甲基化水平能够有效诊断高级别宫颈癌前病变.%Objective To evaluate paired boxed gene 1 (PA X 1) methylation quantitative detection and immunohistochemical diagnostic value of clinical disease for the first high-level cancer gene family. Method The pathological diagnosis as the gold standard, 135 high-risk subtypes of human papilloma virus (HPV) cervical infection were divided into normal group (group Ⅰ, n = 40), low-grade cervical lesions before (groupⅡ, n=44) and high grade cervical lesions before (groupⅢ, n=51) from January 2014 to October 2016 in Sixth People's Hospital Affiliated to Shanghai Jiao Tong University. Collecting cervical tissue samples of each group were used immunohistochemistry (IHC), pyrosequencing detecting PA X 1 methylation and expression level of each group of samples, and statistical analysis, drawing subjects operating characteristic (ROC)curve,to clarify gene methylation level PA X 1 diagnosis of high-grade cervical lesions before.Result PA X 1 methylation level group Ⅰ was (4.53±2.62)%, group Ⅱ was (3.24±2.28)%, Ⅲ group was (13.67±5.38)%, the difference was statistically significant (F = 22.35, P < 0.001). Area under the ROC curve was 0.9216. Experimental results showed that PA X 1 methylation levels can be used for high-grade cervical lesions before diagnosis, the diagnostic threshold of 8.47%, a sensitivity of 86.27% and a specificity of 98.81%.Conclusion Pyrosequencing assay for quantification of cervical tissue methylation levels PA X 1 can effectively diagnose high-level cervical lesions.