Fan Jie, Li Peng, Fang Qigen, Yang Yang, Zhang He, Du Wei, Liu Shanting, Luo Ruihua
Department of Head Neck and Thyroid Surgery, Affiliated Cancer Hospital of Zhengzhou University, Henan Cancer Hospital, Zhengzhou, China.
Department of Nephrology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Front Oncol. 2022 Aug 16;12:807597. doi: 10.3389/fonc.2022.807597. eCollection 2022.
To reveal a novel pathological feature: heterotypic neutrophil-in-tumor structure (hNiT) first discovered in patients with oropharyngeal squamous cell carcinoma (OPSCC), to analyze the prognostic role of hNiT in OPSCC patients and to explore the role of p16 in the formation of hNiT structures.
Clinically, 197 patients were enrolled. Clinicopathological information was extracted and analyzed. All pathologic sections made from primary tumors were re-evaluated by immunohistochemistry and immunostaining. , we cocultured OPSCC cell line SCC-15 with neutrophils to form hNiT structures, which were then subject to fluorescence staining. By RNAi and overexpression techniques, we investigated the role of CDKN2A in the formation of hNiTs. We validated the two techniques by qPCR and Western Blot.
The hNiT as a novel pathological feature was first discovered in the tissues of OPSCC. The FNiT was significantly associated with tumor stage, disease stage, p16 and tumor grade. A total of 119 patients died of the disease, and the 5-year disease-specific survival (DSS) rate was 36%. The median survival time was 52.6 months. In patients with an FNiT<0.5%, the 5-year DSS rate was 40%; in patients with an FNiT>=0.5%, the 5-year DSS was 28%, and the difference was significant (p=0.001). Cox model analysis showed that FNiT along with disease stage, p16 and tumor grade was an independent prognostic factor for DSS. Immunostaining results of p16 expression showed hNiT formation was negatively correlated to p16 in OPSCC as well as in the hNiT formation assays indicated by fluorescent staining. Function assays of CDKN2A implied that reduce CDKN2A promoted the formation of hNiT while elevated CDKN2A impeded the hNiT formation.
The hNiT as a novel pathological feature is associated with the adverse prognosis of OPSCC patients with p16 inhibiting the formation of hNiT structures.
揭示一种首次在口咽鳞状细胞癌（OPSCC）患者中发现的新病理特征：异型肿瘤内中性粒细胞结构（hNiT），分析hNiT在OPSCC患者中的预后作用，并探讨p16在hNiT结构形成中的作用。
临床上，纳入197例患者。提取并分析临床病理信息。对所有原发性肿瘤制作的病理切片进行免疫组织化学和免疫染色重新评估。我们将OPSCC细胞系SCC - 15与中性粒细胞共培养以形成hNiT结构，然后对其进行荧光染色。通过RNAi和过表达技术，我们研究了CDKN2A在hNiT形成中的作用。我们通过qPCR和蛋白质免疫印迹法验证了这两种技术。
hNiT作为一种新的病理特征首次在OPSCC组织中被发现。FNiT与肿瘤分期、疾病分期、p16和肿瘤分级显著相关。共有119例患者死于该疾病，5年疾病特异性生存率（DSS）为36%。中位生存时间为52.6个月。在FNiT<0.5%的患者中，5年DSS率为40%；在FNiT>=0.5%的患者中，5年DSS率为28%，差异有统计学意义（p = 0.001）。Cox模型分析表明，FNiT以及疾病分期、p16和肿瘤分级是DSS的独立预后因素。p16表达的免疫染色结果显示，在OPSCC中hNiT的形成与p16呈负相关，在荧光染色所示的hNiT形成试验中也是如此。CDKN2A的功能试验表明，降低CDKN2A促进hNiT的形成，而升高CDKN2A则阻碍hNiT的形成。
hNiT作为一种新的病理特征与p16抑制hNiT结构形成的OPSCC患者的不良预后相关。
