Laboratory of Immunogenetics, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Recife, Brazil.
Laboratory of Ultrastructure, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Recife, Brazil.
Exp Mol Pathol. 2022 Feb;124:104734. doi: 10.1016/j.yexmp.2021.104734. Epub 2021 Dec 13.
High-grade cervical lesions (HSIL) are associated with the presence of high-risk HPV types, tissue expression of p16, and increased chance of malignant progression, requiring surgical intervention. To improve risk evaluation, we assessed the discriminatory power of the histological findings associated with p16 immunohistochemistry (IHC) staining to classify the low-grade cervical lesion (LSIL) and HSIL.
We collected cervical biopsies from colposcopy-visible lesions and non-affected tissue (adjacent to the lesions) of 62 Brazilian women and labeled them with anti-p16 antibodies. In addition to the observational pattern and labeling to define the latent classes (affected vs. non-affected), a computational tool was used for semi-quantitative analysis of p16 expression. The intensity of staining of the nucleus or cytoplasm was captured using the Gimp 2.10 software. ROC curves were used to determine cutoff values for p16 expression in patients classified as LSIL and HSIL by latent class statistics for each labeling stratum.
p16 nuclear labeling showed the best sensitivity and specificity to discriminate LSIL with low p16 expression (62%) and HSIL with high p16 expression (37%). Many patients whose lesions had intermediate levels of p16 nuclear staining were subsequently stratified according to the expression of p16 in the cytoplasm, indicating that five of 21 LSIL were at risk of progression, and 13 of 41 HSIL at risk of regression.
We suggest a hierarchical analysis, with histology at the first level, followed by a labeling analysis in the nucleus and then in the cytoplasm to increase the accuracy of the HPV cervical lesion stratification.
高级别宫颈病变（HSIL）与高危型 HPV 类型的存在、p16 的组织表达以及恶性进展的可能性增加有关，需要手术干预。为了提高风险评估，我们评估了与 p16 免疫组化（IHC）染色相关的组织学发现对低级别宫颈病变（LSIL）和 HSIL 的区分能力。
我们收集了来自阴道镜可见病变和巴西 62 名女性未受影响组织（病变旁）的宫颈活检，并标记了抗 p16 抗体。除了观察模式和标记以定义潜在类别（受影响与未受影响）外，还使用计算工具对半定量分析 p16 表达。使用 Gimp 2.10 软件捕获核或细胞质染色的强度。使用 ROC 曲线确定通过潜在类别统计为 LSIL 和 HSIL 分类的患者中 p16 表达的截断值，每个标记层。
p16 核标记显示出最佳的敏感性和特异性，可区分低 p16 表达的 LSIL（62%）和高 p16 表达的 HSIL（37%）。许多核 p16 染色水平中等的病变患者随后根据细胞质中 p16 的表达进行分层，表明 21 例 LSIL 中有 5 例有进展风险，41 例 HSIL 中有 13 例有进展风险。
我们建议进行层次分析，首先进行组织学分析，然后进行核标记分析，最后进行细胞质标记分析，以提高 HPV 宫颈病变分层的准确性。
