Department of Pathology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, The Netherlands.
DDL Diagnostic Laboratory, Rijswijk, The Netherlands.
Mod Pathol. 2018 Dec;31(12):1842-1850. doi: 10.1038/s41379-018-0101-z. Epub 2018 Aug 22.
In this study, we evaluate the expression of human papillomavirus E4 protein (marker for the onset of a productive infection) and hypermethylation of host-cell CADM1, MAL, and miR124-2 genes (marker for an advanced, transforming infection) in cervical intraepithelial neoplasia (CIN) and cancer. A total of 115 cervical lesions were categorized by 3 pathologists into no dysplasia, CIN1, CIN2, CIN3, or cancer by classical histomorphological grading criteria, and by an immunoscore (cumulative value: 0-6) grading system based on Ki-67 (score: 0-3) and p16 (score: 0-3) expression. Lesions were immunostained for E4 protein and analyzed for hypermethylation of CADM1, MAL, or miR124-2 genes. Expression of E4 and hypermethylation levels were related to CIN grade based on both classical and immunoscore grading. Hypermethylation increased with severity of the lesion as defined by both classical histomorphological grading and immunoscore criteria, and was always present in carcinomas (22/22). Extensive E4 expression decreased with increasing CIN grade and immunoscore, being most frequent in classically graded CIN1 or in lesions with cumulative immunoscore 1-3 and absent in carcinomas. High-grade lesions (CIN2/3 or immunoscore: 4-6) showed less E4 expression, which was inversely related to an increasing hypermethylation. Extensive E4 expression, as observed in a small proportion of high-grade lesions (6/49 and 8/43, respectively), was mostly associated with a negative methylation marker status (5/6 and 7/8, respectively). Our results illustrate the gradual transition of productive CIN (reflected by extensive E4 expression), to advanced transforming CIN (reflected by extensive hypermethylation) and cancer. Expression patterns of E4 and hypermethylation status of host-cell genes, may be used to identify cervical lesions at risk for cervical cancer, providing a better guidance for clinicians on treatment decisions.
在这项研究中，我们评估了人乳头瘤病毒 E4 蛋白（活跃感染的标志物）的表达和宿主细胞 CADM1、MAL 和 miR124-2 基因的高甲基化（高级别、转化感染的标志物）在宫颈上皮内瘤变（CIN）和癌症中的表达。共有 115 例宫颈病变由 3 位病理学家通过经典组织形态学分级标准分为无发育不良、CIN1、CIN2、CIN3 或癌症，并通过基于 Ki-67（评分：0-3）和 p16（评分：0-3）表达的免疫评分（累积值：0-6）分级系统进行分级。对 E4 蛋白进行免疫染色，并分析 CADM1、MAL 或 miR124-2 基因的高甲基化。E4 的表达和高甲基化水平与基于经典和免疫评分的 CIN 分级相关。随着病变的严重程度增加，高甲基化水平也随之增加，这在经典分级的 CIN1 或累积免疫评分 1-3 的病变中始终存在，在癌中总是存在。广泛的 E4 表达随着 CIN 分级和免疫评分的增加而减少，在经典分级的 CIN1 或累积免疫评分 1-3 的病变中最常见，而在癌中不存在。高级别病变（CIN2/3 或免疫评分：4-6）的 E4 表达较少，这与不断增加的高甲基化呈负相关。广泛的 E4 表达（在少数高级别病变中观察到，分别为 6/49 和 8/43）主要与阴性甲基化标志物状态相关（分别为 5/6 和 7/8）。我们的结果表明，从活跃的 CIN（通过广泛的 E4 表达来反映）到高级别的转化性 CIN（通过广泛的高甲基化来反映）和癌症的逐渐转变。E4 的表达模式和宿主细胞基因的高甲基化状态可用于识别有宫颈癌风险的宫颈病变，为临床医生提供更好的治疗决策指导。
