Background: While histopathology is fundamental to cancer diagnosis, its potential for comprehensive molecular profiling remains underexplored. This study aimed to develop an integrated computational pathology pipeline that predicts gene mutations from histopathology images and explores histology-multi-omics correlations. Methods: We analyzed 874 H&E-stained whole-slide images from 647 head and neck squamous cell carcinoma patients across three international cohorts. The pipeline comprised: 1) Cancer-Net, a segmentation model trained on similar to 200,000 annotated image tiles to identify tumor regions; and 2) GEM-Net, an enhanced teacher-student multiple instance learning framework integrating a Transformer architecture for global slide-level feature aggregation, to capture global context for mutation prediction. Sparse canonical correlation analysis (SCCA) was then applied to explore associations between the image features extracted by GEM-Net and transcriptomic/ proteomic pathways. Results: Cancer-Net achieved high tumor segmentation performance (AUC = 0.995). GEM-Net demonstrated robust mutation prediction (AUC = 0.702-0.876), outperforming CNN and Transformer baselines. A clinically relevant dual-thresholding strategy based on GEM-Net outputs demonstrated the potential to reduce theoretical need for molecular testing by 10-26%. Visual explanations from GEM-Net aligned with immunohistochemical staining patterns. SCCA interpreted the morphological features learned by GEM-Net, revealing associations with molecular pathways such as cell cycle dysregulation in TP53-mutant tumors and tight junction disruption in FAT1-mutant cases. Conclusions: This integrated pipeline enables accurate mutation prediction and provides interpretable links between histopathology and multi-omics data, suggesting potential utility for clinical decision support and cancer biology research.

BackgroundOral squamous cell carcinoma (OSCC) represents a common malignancy characterized by significant morbidity and mortality rates, highlighting the critical necessity for novel therapeutic approaches. Consequently, investigating differentially expressed genes linked to inflammation and pyroptosis may identify potential prognostic biomarkers and therapeutic targets.MethodsTo address this research gap, our study utilized an extensive bioinformatics approach by analyzing the Cancer Genome Atlas Head and Neck Squamous Cell Carcinoma (TCGA-HNSC) dataset through differential expression analysis to identify genomic features associated with OSCC. Subsequent analyses included Gene Ontology and pathway enrichment assessments, along with survival analyses using Cox regression models, to evaluate the prognostic significance of the identified differentially expressed genes. Furthermore, immune infiltration analysis and somatic mutation assessments were conducted to elucidate the relationship between immune cell types and key prognostic genes. Additionally, copy number variation analysis was performed to highlight genomic alterations associated with immune-related and prognostic-related differentially expressed genes(DEGs).ResultsOur analysis identified a total of 3,495 differentially expressed genes, among which 53 immune-related and prognosis-related differentially expressed genes demonstrated a significant correlation with the prognosis of OSCC. Immune infiltration analysis further revealed the presence of 28 immune cell types within OSCC samples, with a notable prevalence of activated CD8 T cells and regulatory T cells, underscoring their association with critical prognostic genes. Additionally, pathway analysis highlighted the activation of cytokine signaling pathways and their associations with processes relevant to systemic lupus erythematosus. A prognostic risk model derived from these findings effectively stratified patients based on overall survival, identifying four key genes—CTSG,HKDC1,PTX3 and SPP1—as crucial prognostic indicators. This analysis may uncover potential prognostic biomarkers and therapeutic targets.ConclusionThis study established a novel OSCC prognostic risk model based on inflammation- and pyroptosis-related interactive genes. CTSG, HKDC1, PTX3 and SPP1 were validated as independent prognostic biomarkers, and the risk score was closely associated with tumor microenvironment features, metabolic activity, and therapeutic sensitivity. This work may provides substantial references for the exploration of novel biomarkers for OSCC treatment and facilitates clinical decision-making.

[This corrects the article DOI: 10.3389/fonc.2022.807597.].

Hypopharyngeal squamous cell carcinoma (HSCC) is a highly invasive and fatal tumor with a poor prognosis in head and neck tumors. It is urgent to further study the molecular mechanism of HSCC progression and identify new effective therapeutic targets. Cell division cycle-related protein 3 (CDCA3) was reported overexpressed in several cancers and involved in tumor progression. However, the biological role of CDCA3 and its potential mechanism in HSCC remain undetermined. Reverse transcription quantitative polymerase chain reaction (RT-PCR) and immunohistochemistry were used to detect the expression levels of CDCA3 in HSCC tissue and matched peritumoral tissue. The effects of CDCA3 on cell proliferation, invasion, and migration were explored using the Celigo image cytometry assay, MTT assay, flow cytometric analysis, cell invasion, and migration assays. The results showed that CDCA3 was upregulated in HSCC tissue and FaDu cell line. Knockdown of CDCA3 inhibited the proliferation, invasion, and migration of FaDu cells and promoted apoptosis of FaDu cells. Furthermore, knockdown of CDCA3 blocked the cell cycle in the G0/G1 phase. Mechanistically, CDCA3 may play a role in tumor progression of HSCC through the Akt/mTOR signaling pathway. In summary, these results suggest that CDCA3 serves as an oncogene in HSCC and may be used as a prognostic indicator and a potential therapeutic target for HSCC.

Papillary thyroid carcinoma (PTC) is one of the most common thyroid carcinomas. The gross extrathyroidal extension and extensive metastases of PTC lead to high rates of recurrence and poor clinical outcomes. However, the mechanisms underlying PTC development are poorly understood. In this study, using single-cell RNA sequencing, the transcriptome profiles of two PTC patients were addressed, including PTC1 with low malignancy and good prognosis and PTC2 with high malignancy and poor prognosis. We found that epithelial subcluster Epi02 was the most associated with the malignant development of PTC cells, with which the fold change of Chitinase 3-like 1 (CHI3L1) is on the top of the differentially expressed genes between PTC1 and PTC2 ( P < 0.001). However CHI3L1 is rarely investigated in PTC as far. We then studied its role in PTC with a series of experiments. Firstly, qRT-PCR analysis of 14 PTC patients showed that the expression of CHI3L1 was positively correlated with malignancy. In addition, overexpression or silencing of CHI3L1 in TPC-1 cells, a PTC cell line, cultured in vitro showed that the proliferation, invasion, and metastasis of the cells were promoted or alleviated by CHI3L1. Further, immunohistochemistry analysis of 110 PTC cases revealed a significant relationship between CHI3L1 protein expression and PTC progression, especially the T ( P < 0.001), N ( P < 0.001), M stages ( P = 0.007) and gross ETE ( P < 0.001). Together, our results prove that CHI3L1 is a positive regulator of malignant development of PTC, and it promotes proliferation, invasion, and metastasis of PTC cells. Our study improves understanding of the molecular mechanisms underlying the progression of PTC and provides new insights for the clinical diagnosis and treatment of PTC.

In recent years, the incidence of thyroid cancer has increased significantly. Most of the cases are differentiated thyroid cancer, which is characterized by a good prognosis. However, 15% of patients still have persistent or recurrent disease after initial treatment, and those with locally advanced or metastatic cancer are not cured with established treatments and at risk of death. The treatment of advanced thyroid cancer is still controversial at home and abroad, but it tends to targeted therapy and immunotherapy. With the in-depth understanding of the molecular pathogenesis of thyroid cancer, a variety of new targeted therapies have been approved for advanced thyroid cancer. The 2021 guidelines of the Chinese Society of Clinical Oncology (CSCO) and the 2022 guidelines of the European Society of Oncology (ESMO) regard targeted therapy as a level I recommendation for the treatment of advanced thyroid cancer. This article reviews the new progress in clinical treatment of advanced thyroid carcinoma.

[This corrects the article DOI: 10.3389/fendo.2021.716082.].

BackgroundTo explore the metabolic differences of follicular thyroid carcinoma (FTC) by metabonomics, to find potential biomarkers for the diagnosis of FTC, and to explore the pathogenesis and diagnosis and treatment strategies of FTC.MethodThe metabonomics of 15 patients with FTC and 15 patients with follicular thyroid nodules(FTN) treated in Henan Cancer Hospital were analyzed by liquid chromatography-mass spectrometry (LC-MS).ResultsThe analysis showed that the metabolite profiles of FTC tissues could be well distinguished from those of control tissues, and 6 kinds of lipids were identified respectively, including lysophosphatidic acid(LysoPA) [LysoPA(0:0/18:0),LysoPA(0:0/18:2(9Z,12Z)],LysoPA[20:4(8Z,11Z,14Z,17Z)/0:0)]; phosphatidic acid(PA) [PA(20:3(8Z,11Z,14Z)/0:0),PA(20:4(5Z,8Z,11Z,14Z)/0:0),PA(20:5(5Z,8Z,11Z,14Z,17Z)/0:0)]; lysophosphatidylcholine(LPC) [LPC(18:1),LPC(16:0),LPC[16:1(9Z)/0:0],LPC(17:0),LPC[22:4(7Z,10Z,13Z,16Z),LPC(20:2(11Z,14Z); phosphatidylcholine(PC)(PC(14:0/0:0),PC(16:0/0:0); sphingomyelin(SM) (d18:0/12:0); fatty acid(FA)(18:1(OH3)]. There are 2 kinds of amino acids, including L-glutamate,L-glutamine.There are 3 other metabolites, including retinol,flavin adenine dinucleotide,androsterone glucuronide.Lipid metabolites are the main metabolites in these metabolites.The metabolic pathways related to FTC were analyzed by KEGG and HMDB, and 9 metabolic pathways were found, including 4 amino acid related metabolic pathways, 1 lipid metabolic pathways and 4 other related pathways.ConclusionThere are significant differences in many metabonomic characteristics between FTC and FTN, suggesting that these metabolites can be used as potential biomarkers. Further study found that LysoPA and its analogues can be used as biomarkers in the early diagnosis of FTC.It may be related to the abnormal metabolism of phospholipase D (PLD), the key enzyme of LysoPA synthesis caused by RAS pathway. At the same time, it was found that the metabolic pathway of amino acids and lipids was the main metabolic pathway of FTC. The abnormality of LysoPA may be the cause of follicular tumor carcinogenesis caused by lipid metabolic pathway.

ObjectiveTo 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. MethodsClinically, 197 patients were enrolled. Clinicopathological information was extracted and analyzed. All pathologic sections made from primary tumors were re-evaluated by immunohistochemistry and immunostaining. In vitro, 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. ResultsThe 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 in vitro indicated by fluorescent staining. Function assays of CDKN2A implied that reduce CDKN2A promoted the formation of hNiT while elevated CDKN2A impeded the hNiT formation. ConclusionThe hNiT as a novel pathological feature is associated with the adverse prognosis of OPSCC patients with p16 inhibiting the formation of hNiT structures.

ObjectivesOur aim was to describe our experience in using apatinib as treatment for radioiodine-refractory differentiated thyroid carcinoma (RAIR-DTC).MethodsForty-seven patients undergoing apatinib treatment for RAIR-DTC were prospectively enrolled in this study. The study endpoints were objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS), overall survival (OS), and rate of adverse events.ResultsNo patients achieved complete response, while 36 (76.6%) and 8 (17.0%) patients achieved partial response and stable disease, respectively. The ORR and DCR were 76.6% and 93.6%, respectively. The median PFS and OS were 18 and 59 months, respectively. A total of 91 adverse events occurred, of which 21 were graded as grade 3 or higher. There were no drug-related deaths.ConclusionsApatinib has distinct anti-RAIR-DTC efficacy in terms of ORR, PFS, and OS and has a favorable safety profile. It is a feasible treatment option for RAIR-DTC.

目的 探讨甲状腺癌患者行甲状腺癌根治术和颈部淋巴结清扫术前口服橄榄油预防术后乳糜漏的效果.方法 选择2013年1月至2019年12月河南省肿瘤医院收治的2627例甲状腺癌患者为研究对象,所有患者行甲状腺癌根治术和左侧颈部淋巴结清扫术,按术前是否口服橄榄油将患者分为观察组(n=1348)和对照组(n=1279).观察组患者术前6 h口服橄榄油1 mL·kg-1,对照组患者术前未口服任何油剂或乳剂制剂.对2组患者术中胸导管辨识率、术后乳糜漏发生率及乳糜漏患者术后日最大引流量、总引流量及引流时间进行比较.结果 所有患者均顺利完成手术,术中术后无死亡病例.观察组和对照组患者术中胸导管辨识率分别为91.17％(1229/1348)、65.36％(836/1279),观察组患者术中胸导管辨识率显著高于对照组(χ2=259.951,P<0.05).观察组和对照组患者乳糜漏发生率分别为1.85％(25/1348)、3.44％(44/1279),观察组患者乳糜漏发生率显著低于对照组(χ2=6.451,P<0.05).观察组乳糜漏患者日最大引流量和总引流量显著少于对照组,引流时间显著短于对照组(P<0.05).结论 甲状腺癌患者行甲状腺癌根治术和颈部淋巴结清扫术前口服橄榄油可以显著提高胸导管辨识率,降低乳糜漏发生率.

Objective:To explore the application value of high frequency ultrasound and ultrasound-guided fine needle aspiration biopsy（US-FNAB） in the diagnosis of papillary thyroid microcarcinoma（PTMC）, and to compare the characteristics and value of the two methods, so as to find a more convenient and non-invasive diagnostic method of PTMC, reduce unnecessary puncture and operation. Methods:The data of 190 postoperative pathologically confirmed PTMC patients admitted to Henan Province Cancer Hospital and Henan Provincial Hospital from January to June 2020 were retrospectively analyzed, with a total of 305 nodules, including 198 PTMC nodules and 107 benign thyroid nodules（BTN）. According to the postoperative pathological results, they were divided into groups, and the relationship between the ultrasound appearance of the nodules and whether the cervical lymph nodes could be explored and PTMC was analyzed by chi-square test and logistic regression, and its diagnostic value was evaluated. The Kappa consistency test was used to analyze the consistency between ultrasound, FNAB and surgical pathological diagnosis results. The accuracy, sensitivity and specificity of high-frequency ultrasound and US-FNAB were compared, and the ROC curve was used to calculate the maximum area under the curve to evaluate its effectiveness. Results:The chi-square test showed that there were statistically significant differences in the morphology, margin, internal echo, echo uniformity, calcification, aspect ratio, blood flow signal, and whether the cervical lymph nodes can be detected and other ultrasound signs between the PTMC group and the BTN group. Logistic regression analysis showed that irregular shape, unclear edges, internal hypoechoic, intranodular calcification are independent risk factors for PTMC. By consistency test, the consistency between high-frequency ultrasound, US-FNAB examination and surgical pathological diagnosis was good, Kappa value was 0.802 and 0.893（P<0.05）. Each nodule was examined by high-frequency ultrasound, and the diagnostic sensitivity, specificity, accuracy and AUC were 95.45%, 83.18%, 91.15% and 0.877 respectively. US-FNAB was performed on 189 of 305 thyroid nodules, and the diagnostic sensitivity, specificity, accuracy and AUC were 96.03%, 93.65%, 95.24% and 0.948 respectively. Conclusion:High frequency ultrasonic features such as internal hypoechoic, calcification in the nodules, unclear edges, and irregular morphology are of high value for the diagnosis of PTMC. Through data analysis, both high-frequency ultrasound and US-FNAB examination have high diagnostic value for PTMC. Compared with US-FNAB, high-frequency ultrasound has the advantages of low examination cost, non-invasive, simple operation and so on. For some patients with PTMC who do not have high risk factors, ultrasound can be used to actively monitor disease progression to avoid some unnecessary surgery.

大多数甲状腺乳头状癌经过适当手术治疗后预后良好,然而局部侵袭使手术治疗复杂化,并且有很高的术后复发率和死亡率[1].7％～16％的甲状腺癌可直接侵犯喉、气管、食管、喉返神经以及颈部大血管等周围组织,也是分化型甲状腺癌最常见的致死原因之一[2].因此,局部控制甲状腺浸润癌是一个重要的临床问题.甲磺酸阿帕替尼是以血管内皮生长因子受体2 (vascular endothelial growth factor receptor 2,VEGFR2)为靶点的小分子酪氨酸激酶抑制剂(tyrosine kinase inhibitor,TKI),已被我国食品药品监督管理总局(China Food and Drug Adminis-tration,CFDA)批准上市[3].目前,关于阿帕替尼作为局部晚期甲状腺癌的新辅助治疗的相关报道比较罕见.本研究报道1例局部晚期甲状腺癌患者,术前细针穿刺细胞病理学确诊为甲状腺乳头状癌.由于首次就诊时肿瘤的根治性切除较困难,因此在术前应用靶向药物甲磺酸阿帕替尼治疗11周后,肿瘤大小明显缩小,停药2周后进行了根治性切除手术,术后病理确诊为经典型乳头状癌,患者术后恢复顺利,随诊无复发迹象.

Objective To study the relationship between DNA double-strand break (DSB) repair gene mutations and the risk of papillary thyroid microcarcinoma (PTMC). Methods One hundred patients with PTMC or benign thyroid nodules (BTNs) at Henan Cancer Hospital were retrospectively analyzed. The DSB repair capacity of peripheral blood T lymphocytes in the two groups was assessed by flow cytometry. Data were compared using Student’s t-test to evaluate the relationship between DSB repair capacity and the risk of PTMC. Factors influencing DSB repair capacity were analyzed by multivariate logistic regression analysis. The relationship between PTMC and DSB repair capacity was analyzed by univariate analysis. Targeted next-generation DNA sequencing was applied to screen and analyze DSB repair genes related to PTMC. Results The DSB repair capacity was 31.30% in the PTMC group and 44.40% in the BTN group, with that of the former being significantly lower ( P < 0.05). Multivariate logistic regression analysis of age, sex, obesity status, radiation and other factors showed that radiation exposure was positively correlated with reduced DSB repair capacity( OR = 3.642; 95% CI 1.484–8.935, P = 0.020). Moreover, univariate analysis showed that a reduction in DSB repair capacity was a risk factor for PTMC ( OR = 2.333; 95% CI 1.027–5.300, P = 0.043).Targeted next-generation DNA sequencing was performed on the DSB repair genes discovered, and those that were mutated in association with PTMC were Rad50 and FANCA; Rad51 mutations were related to BTN. Conclusion Radiation exposure is positively associated with induced DSB repair gene mutations, which may cause a reduced capacity for DSB repair and eventually lead to PTMC.

ObjectiveTo analyze the incidence and risk factors for lateral lymph node metastases (LNMs) in T1a papillary thyroid carcinoma (PTC) with a focus on tumor location and size.Materials and MethodsThe incidence of lateral LNM in 345 cases of T1a PTC was retrospectively analyzed. Univariate and multivariate analyses were performed to assess the relationships between lateral LNM and clinicopathological characteristics.ResultsThe incidence of skip metastasis to lateral LNM in T1a PTC located in the upper lobe was 12.1% (8/66). Logistic regression analysis indicated tumor size >5 mm (OR = 5.04, 95% CI = 1.79 to 14.18, P = 0.002), upper lobe location (OR = 7.68, 95% CI = 3.05–19.34, P < 0.001) and the number of central neck LNM (<2: OR = 24.79, 95% CI = 8.23–74.60, P < 0.001; ≥2: OR = 4.99, 95% CI = 1.95–12.73, P < 0.001) were independently associated with lateral LNM. Comparing the lateral and central LNM stratification based on tumor location revealed that both the incidences of lateral (33.3%) and central (30.3%) LNM of T1a PTC located in the upper lobe were higher than those of T1a PTC located in the middle and lower lobes. Of T1a PTC located in the upper lobe, the incidence of lateral LNM was 33.3% (22/66), which was higher than that [30.3% (20/66)] of central LNM. This finding is reversed in all T1a PTC cases and T1a PTC cases with tumor located in the middle and lower lobes.ConclusionA particularly high likelihood of lateral LNM was observed in T1a PTC patients with tumor located in the upper lobe of the thyroid gland, especially the tumor >5 mm in size, which could be considered a risk factor for lateral LNM in the clinical management of T1a PTC.

目的:探索与下咽鳞癌生长、转移及预后相关的分子标记物,为开发下咽鳞癌新的疗法提供理论依据.方法:分析Oncomine肿瘤芯片数据库中细胞分裂周期相关蛋白3(cell division cycle associated protein 3,CDCA3)在头颈部鳞癌及癌旁组织中的表达差异,检测下咽鳞癌及癌旁组织中CDCA3的表达,分析其表达水平与下咽鳞癌患者临床病理特征及预后的关系.结果:通过分析Oncomine肿瘤芯片数据库中CDCA3的DNA拷贝数,发现在肿瘤基因图谱(TCGA)数据集中,相比全血及正常头颈部组织,CDCA3的DNA拷贝数在头颈鳞癌组织中显著上调(P<0.05).下咽鳞癌组织中CDCA3的mRNA相对表达水平较癌旁正常黏膜表达水平升高(P<0.05).CDCA3的蛋白表达水平与肿瘤大小、淋巴结转移及TNM分期等临床病理特征相关(P<0.05).Kaplan-Meier生存曲线分析显示CDCA3阳性表达组患者的总生存期显著低于CDCA3阴性表达组(P<0.05).结论:CDCA3在下咽鳞癌组织中高表达,与下咽鳞癌患者原发肿瘤大小、TNM分期以及淋巴结转移情况密切相关,是影响患者预后的独立危险因素.

女,49岁.2018年3月16日因间断头痛5年就诊.磁共振成像(MRI)检查显示:右额叶占位,水肿明显,考虑为转移瘤(图1).2018年3月18日出现昏迷,急诊行右额叶肿物切除术+去骨瓣减压术.术后病理诊断:右额叶转移性乳头状癌,免疫组化检查支持诊断甲状腺乳头状癌转移.免疫组化:CK19(+),CD56(-),TG(+),TTF-1(+), 34BE12(-),CK(+),GFAP(-),S-100(-),SyN(-),Ki-67约8％(图2).

Purpose: Our goal was to analyze the feasibility of submandibular gland (SMG) preservation in cT1-2N0 floor of the mouth (FOM) squamous cell carcinoma (SCC) patients. Methods: Patients with cT1-2N0 FOM SCC were retrospectively enrolled and divided into two groups according to the management of the SMG. Level 1b tissues were divided into six groups according to their location with respect to the SMG. The Kaplan-Meier method was used to calculate the locoregional control (LRC) and disease-specific survival (DSS) rates. A Cox model was used to determine the independent risk factors. Results: Twenty-nine patients underwent SMG-preserving neck dissection, and lymph node metastasis occurred in the superior group in 3 of the 37 dissections with a prevalence of 8.1% and in the anterior group in 2 of the 37 dissections with a prevalence of 5.4%. In patients without SMG preservation, lymph node metastasis occurred in the superior group in 7 of the 137 dissections with a prevalence of 5.1% and in the anterior group in 6 of the 137 dissections with a prevalence of 4.4%. The only pattern of SMG involvement was invasion by positive lymph nodes. The 5-year LRC rates for patients with SMG preservation and patients with SMG excision were 84 and 73%, respectively, and the difference was not significant (p = 0.239). The 5-year DSS rates for patients with SMG preservation and patients with SMG excision were 88 and 84%, respectively, and the difference was not significant (p = 0.524). Conclusions: In early-stage FOM SCC patients, SMG involvement is rare, the most common metastatic site in level 1b is the superior group, and SMG preservation does not decrease the LRC or DSS rates. Therefore, the findings suggest that there might be high feasibility of SMG-preserving neck dissection in cT1-2N0 FOM SCC.

Objective To analyze the role of frequency of heterotypic neutrophil-in-tumor structure (FNiT) in the prognosis of patients with buccal mucosa squamous cell carcinoma (BMSCC). Methods In vitro, we cocultured BMSCC cell line-H157 with neutrophils to form heterotypic neutrophil-in-tumor structures, which were then subject to fluorescence staining. Clinically, 145 patients were retrospectively enrolled. Associations between FNiT and clinicopathological variables including age, sex, smoking history, drinking history, betel nut chewing, tumor stage, node stage, metastasis, disease stage, lymphovascular invasion, extranodal extension, perineural invasion, and tumor grade were analyzed by chi-square test, and the main endpoints of interest were recurrence-free survival (RFS) and disease-specific survival (DSS) which were analyzed by the Kaplan-Meier method and Cox model. Results Fluorescent staining results of typical heterotypic neutrophil-in-tumor structure showed that well-differentiated H157 cells had a stronger ability to internalize more neutrophils than poorly-differentiated H157 cells, with the latter often internalizing only one neutrophil or nothing. The mean FNiT was 4.2‰, with a range from 2.3‰ to 7.8‰. A total of 80 patients relapsed and 84 patients died of the disease. The 5-year RFS and DSS rate was 42% and 42%, respectively. Patients with an FNiT≥4.2‰ had a significantly higher risk for locoregional recurrence and cancer-caused death than those with an FNiT<4.2‰ (p=0.001 and p<0.001, respectively). The FNiT alone was independently significant in predicting poor RFS, and the FNiT along with tumor grade was an independent predictor for DSS. Conclusion The FNiT as a novel predictor is significantly negatively associated with both the RFS and DSS of patients with BMSCC.