Recently, the World Health Organization (WHO) recognized human papilloma virus (HPV)-independent invasive cervical squamous cell carcinoma (SCC) without recognizing the existence of precursor lesions. This is a detailed characterization of 3 preinvasive lesions and 6 invasive SCC negative for HPV-DNA (32 genotypes), HPV-mRNA (14 genotypes) and genomic HPV sequencing. We evaluated histologic features, expression of p16^ink4a, p53, CK7, and CK17, aberrations in 50 cancer genes and chromosomal copy number variations. HPV-negative preinvasive lesions were extensive basaloid or highly differentiated keratinizing intraepithelial proliferations of 3 to 20 cell layers thickness, partly with prominent cervical gland involvement. Overall, 2/3 intraepithelial lesions and the in situ component of 1/6 SCC showed p16^ink4a block staining, while 1/6 in situ component revealed heterogenous p16^ink4a staining. All invasive components of keratinizing SCC were p16^ink4a-negative. Preinvasive and invasive SCC showed inconsistent CK7 and CK17 staining. Nuclear p53 overexpression was restricted to the TP53 gene mutated SCC. The highly vascularized peritumoral stroma showed a dense inflammatory infiltrate including plasma cells and intratumoral and peritumoral eosinophilic granulocytes. Inconsistent somatic gene mutations (PIK3CA, STK11, TP53, SMARC2B, and GNAS) occurred predominantly in nonhotspot locations at low mutational frequency in 3/6 SCC. Consistent aberrations included the pathogenic (angiogenic) germline polymorphism Q472H in the KDR gene (7/9 patients), and chromosome 3q gains (4/9 patients). In conclusion, HPV-negative intraepithelial cervical precancerous lesions exist, either as highly differentiated keratinized intraepithelial proliferations reminiscent of differentiated vulvar intraepithelial neoplasia, or undifferentiated basaloid intraepithelial lesions with occasional p16^ink4a block staining resembling high-grade squamous intraepithelial lesion. Gains of chromosome 3q, angiogenic germline variants the inflammatory infiltrate may contribute to progression of HPV-negative cervical carcinogenesis.

最近，世界卫生组织（WHO）承认了不依赖人乳头状瘤病毒（HPV）的浸润性宫颈鳞状细胞癌（SCC），但没有认识到前驱病变的存在。这是 3 个浸润前病变和 6 个浸润性鳞状细胞癌的详细特征，HPV-DNA（32 种基因型）、HPV-mRNA（14 种基因型）和基因组 HPV 测序均为阴性。我们评估了组织学特征、p16 ^ink4a、p53、CK7 和 CK17 的表达、50 个癌症基因的畸变以及染色体拷贝数变异。HPV阴性的浸润前病变是广泛的基底细胞样或高度分化的角化上皮内增殖，厚度为3至20个细胞层，部分伴有明显的宫颈腺受累。总体而言，2/3 上皮内病变和 1/6 SCC 的原位成分显示 p16 ^ink4a块染色，而 1/6 原位成分显示异质 p16 ^ink4a染色。角化性鳞状细胞癌的所有侵袭性成分均为 p16 ^ink4a阴性。侵袭前和侵袭性 SCC 显示不一致的 CK7 和 CK17 染色。核p53过度表达仅限于TP53基因突变的SCC。高度血管化的瘤周间质显示出致密的炎症浸润，包括浆细胞以及瘤内和瘤周嗜酸性粒细胞。不一致的体细胞基因突变（PIK3CA、STK11、TP53、SMARC2B 和 GNAS）主要发生在 3/6 SCC 中突变频率较低的非热点位置。一致的畸变包括 KDR 基因中的致病性（血管生成）种系多态性 Q472H（7/9 名患者）和染色体 3q 增益（4/9 名患者）。总之，存在 HPV 阴性宫颈上皮内癌前病变，要么是高度分化的角化上皮内增生，让人想起分化的外阴上皮内瘤变，要么是未分化的基底样上皮内病变，偶尔有 p16 ink4a 块染色，类似于高级鳞状上皮内^病变。3q 染色体、血管生成种系变异和炎症浸润的增加可能会导致 HPV 阴性宫颈癌的进展。