FADD amplification is associated with CD8+ T-cell exclusion and malignant progression in HNSCC.

OBJECTIVE: This study aimed to clarify the relationship between FADD amplification and overexpression and the tumor immune microenvironment. METHODS: Immunohistochemical staining and bioanalysis were used to analyze the association between FADD expression in tumor cells and cells in tumor microenvironment. RNA-seq analysis was used to detect the differences in gene expression upon FADD overexpression. Flow cytometry and multicolor immunofluorescence staining (mIHC) were used to detect the differences in CD8+ T-cell infiltration in FADD-overexpressed cells or tumor tissues. RESULTS: Overexpression of FADD significantly promoted tumor growth. Cells with high FADD expression presented high expression of CD276 and FGFBP1 and low expression of proinflammatory factors (such as IFIT1-3 and CXCL8), which reduced the percentage of CD8+ T cells and created a "cold tumor" immune microenvironment, thus promoting tumor progression. In vivo and in vitro experiment confirmed that tumor tissues with excessive FADD expression exhibited CD8+ T-cell exclusion in the microenvironment. CONCLUSION: Our preliminary investigation has discovered the association between FADD expression and the immunosuppressive microenvironment in HNSCC. Due to the high frequent amplification of the chromosomal region 11q13.3, where FADD is located, targeting FADD holds promise for improving the immune-inactive state of tumors, subsequently inhibiting HNSCC tumor progression.

Role of T-box genes in cancer, epithelial-mesenchymal transition, and cancer stem cells.

Sharing a common DNA binding motif called T-box, transcription factor T-box gene family controls embryonic development and is also involved in cancer progression and metastasis. Cancer metastasis shows therapy resistance and involves complex processes. Among them, epithelial-mesenchymal transition (EMT) triggers cancer cell invasiveness and the acquisition of stemness of cancer cells, called cancer stem cells (CSCs). CSCs are a small fraction of tumor bulk and are capable of self-renewal and tumorsphere formation. Recent progress has highlighted the critical roles of T-box genes in cancer progression, EMT, and CSC function, and such regulatory functions of T-box genes have emerged as potential therapeutic candidates for cancer. Herein we summarize the current understanding of the regulatory mechanisms of T-box genes in cancer, EMT, and CSCs, and discuss the implications of targeting T-box genes as anticancer therapeutics.

Level IV neck dissection in cN0 HPV-negative oropharyngeal squamous cell carcinoma: a retrospective cohort study.

BACKGROUND: As opposed to observation of the neck, elective neck dissection has a survival benefit for cN0 oropharyngeal squamous cell carcinoma (OPSCC). However, there are limited date on level IV neck dissection in human papillomavirus (HPV)-negative OPSCC because most earlier studies did not stratify by P16 or HPV status. Thus, whether to exclude level IV from selective dissection (SND) of cN0 HPV-negative OPSCC remains controversial. METHODS: In this single-center retrospective cohort study, disease-free survival (DFS) was estimated as the primary endpoint for 124 cN0 HPV-negative OPSCC patients who received SND of levels I-III (Group A) and I-IV (Group B). Overall survival (OS) and disease-specific survival (DSS) were considered secondary endpoints. RESULTS: For the entire cohort, the 5-year DFS rates of Groups A and B were 55.0% and 60.1%, respectively. Five-year OS rates were 58.9% and 61.5%, and 5-year DSS rates were 74.0% and 64.8%, respectively. Group B did not show higher 5-year DFS, OS, or DSS than Group A. CONCLUSIONS: This retrospective cohort study validated that in cN0 HPV-negative OPSCC, SND including level IV does not have substantial benefits regarding DFS, OS or DSS.

Excision margin of T3 basal cell carcinoma in maxillofacial area: A retrospective cohort study of Asians.

OBJECTIVES: Basal cell carcinoma is the most common skin cancer worldwide. Surgical excision is considered as the mainstay of treatment, while the evidence of excision margin in advanced stage is lacking, especially in maxillofacial area. MATERIALS AND METHODS: We conducted a 2-center retrospective cohort study. Disease-free survival rate was estimated for 116 Asian patients with T3 basal cell carcinoma in maxillofacial area who received stand surgical excision with margin of 3-5 mm (Group A), 6-9 mm (Group B), and 10-15 mm (Group C). RESULTS: For the entire cohort, five-year disease-free survival rates of Groups A, B, and C were 82.1%, 93.5%, and 92.4%, respectively. When compared with Group B, Group A was correlated with lower disease-free survival rate (HR 5.48, p = .04), and Group C was not associated with different disease-free survival rate (HR 0.85, p = .62). Perineural invasion (p = .006) and pathologic subtypes of infiltrative basal cell carcinoma (p = .01) were independent prognosticator for disease-free survival rate. CONCLUSIONS: This multicenter cohort study validated that T3 basal cell carcinoma Asian patients of maxillofacial area treated with excision margin of 6-9 mm had a substantial benefit of disease-free survival rate and skin conservation.

Salvage surgery for patients with residual/persistent diseases after improper or insufficient treatment of oral squamous cell carcinoma: can we rectify these mistakes?

BACKGROUND: Patterns of failure after treatment of oral and squamous cell carcinomas (OSCC) are diversified, with recurrences being one of the common causes. A special group of patients are sometimes encountered in the outpatient clinic for improper or insufficient initial treatment with reports of positive margins, implying residual/persistent diseases. The question of whether these patients can be surgically salvaged remain unanswered. METHODS: A retrospective study was performed between January 2013 and December 2017 for patients with residual or rapid recurrent (within 3 months) OSCCs, who received salvage surgeries in our institution. The patients with residual/persistent OSCCs were those with microscopic or macroscopic positive surgical margins, while those with rapid recurrent OSCCs were those with close or negative margins, but unabated painful symptoms right after treatment. Both clinicopathological and prognostic variables were analyzed. The focus was also directed towards lessons for possible initial mistakes, resulting in these residual/persistent diseases. RESULTS: Of 103 patients, 68 (66%) were men, with mean age of 56.3 years. The overall survival reached 60.2%. Regarding the primary OSCC status, most of our patients (n = 75, 72.8%) were diagnosed with ycT2-3 stages. Besides, most patients were found with macroscopic residual diseases (52.4%) before our salvage surgery. The sizes of the residual/persistent OSCCs were generally under 4 cm (87.3%) with minimally residual in 21 (20.4%). Among all the variables, primary T stage (p = 0.003), and residual lesion size (p < 0.001) were significantly associated with the prognosis in multivariate analysis. Though the causes for the initial surgical failure were multifactorial, most were stemmed from poor planning and unstandardized execution. CONCLUSIONS: Cases with residual/persistent OSCCs were mostly due to mistakes which could have been avoided under well-round treatment plans and careful surgical practice. Salvage surgery for cases with smaller residual/persistent OSCCs is still feasible with acceptable outcomes.

The multifaceted roles of nucleophagy in cancer development and therapy.

Autophagy is an evolutionarily conserved process in which the cell degrades its own components and recycles the biomolecules for survival and homeostasis. It is an important cellular process to eliminate pathogens or damaged organelles. Nucleophagy, also termed as nuclear autophagy, is a more recently described subtype of autophagy, in which nuclear components, such as nuclear lamina and DNA, are to be degraded. Nucleophagy plays a double-facet role in the development of cancer. On one hand, the clearance of damaged DNA or nuclear structures via autophagic pathway is crucial to maintain nuclear integrity and prevent tumorigenesis. On the other hand, in later stages of tumor growth, nucleophagy may facilitate cancer cell survival and metastasis in the nutrient-depleted microenvironment. In this review, we discuss the relationship between nucleophagy and cancer along with potential intervention methods to target cancer through manipulating nucleophagy. Given the known observations about nucleophagy, it could be promising to target different nuclear components during the processes of nucleophagy, especially nuclear lamina. Further research on investigating the role of nucleophagy in oncological context could focus on dissecting its remaining molecular pathways and their connection to known tumor suppressors.

HSP27 associates with epithelial-mesenchymal transition, stemness and radioresistance of salivary adenoid cystic carcinoma.

Epithelial-mesenchymal transition (EMT) has been shown to associate with cancer stem cells and radioresistance. However, it is obscure whether EMT itself or specific EMT regulators play causal roles in these properties of salivary adenoid cystic carcinoma (SACC). Here, we exhibited that overexpression of HSP27 drove the migration and invasion, induced EMT, as well as mediated TGF-ß1-induced EMT in SACC cells, accompanying the up-regulation of Snail1 and Prrx1. Conversely, HSP27 silencing reduced the migration and invasion and contributed to MET of SACC cells. HSP27 indirectly down-regulates the expression of E-cadherin through activating Snail1 and Prrx1 expressions. Overexpression of Snail1 or Prrx1 restored the migration and invasion in HSP27 knockdown cells. Enforced expression of HSP27 enhanced colony formation, CD133+ /CD44+ population and radioresistance of SACC cell lines. In addition, HSP27 expression was positively associated with radioresistance and poor prognosis of SACC patients as well as with the expression of Prrx1 or Snail1 in SACC tissues. The data confirm an important function for HSP27 in SACC progression through regulating EMT and stemness, and they imply the possible association between EMT and radioresistance of SACC.

Long noncoding RNAs: emerging regulators of tumor angiogenesis.

Long noncoding RNAs (lncRNAs) participate in multiple biological processes especially human diseases, of which, tumor seems to be one of the most significant. Angiogenesis has been deemed to have a pivotal role in a series of tumor biological behaviors in tumorigenesis, progression and prognosis. Emerging evidences suggested that lncRNAs are involved in tumor angiogenesis and lncRNAs have already been verified to be potential biomarkers and promising therapeutic targets. This review summarized emerging angiogenesis-related lncRNAs, discussed their mechanisms interacting with cytokines, cancer stem cells, miRNAs and tumor hypoxia microenvironment, and demonstrated if lncRNAs could be new candidate targets of antiangiogenesis therapy.

The role of tumor microenvironment in collective tumor cell invasion.

For many cancer types, cancer cells invade into surrounding tissues by collective movement of cell groups that remain connected via cell-cell junctions. This migration is completely distinguished from single-cell migration, in which cancer cells disrupt the tight intercellular junctions and gain a mesenchymal phenotype. Recently, emerging evidence has revealed that collective cell invasion depends on not only cell-intrinsic mechanisms but also on extracellular mechanisms by bidirectional interplay between the tumor cell and the tumor environment. Herein, in this review we discuss the role and underline mechanisms of tumor microenvironment in collective tumor cell invasion, particularly focusing on extracellular matrix remodeling and cross-talk between tumor and stromal cells.

Emission Computed Tomography for the Diagnosis of Mandibular Invasion by Head and Neck Cancers: A Systematic Review and Meta-Analysis.

PURPOSE: To detect the diagnostic efficacy of emission computed tomography (ECT) in detecting mandibular invasion caused by head and neck cancers. MATERIALS AND METHODS: Thirteen databases were searched electronically to retrieve studies for inclusion and a manual search also was conducted. Study inclusion, data extraction, and quality assessment were completed by 2 reviewers independently. Meta-DiSc 1.4 and STATA 11.0 were used to conduct the meta-analysis. RESULTS: Seventeen studies involving 668 participants were included. One study had a low risk of bias, 2 had a high risk, and the rest had unclear risk. Meta-analysis showed that for the diagnosis of mandibular invasion single-photon ECT (SPECT) had a mean sensitivity (SEN) of 0.96, a mean specificity (SPE) of 0.66, an area under the curve (AUC) of 0.8989, and a Q* (point on the summary reviewer operator characteristic curve when SEN equaled SPE) of 0.8300. Positron emission tomography combined with computed tomography (PET/CT) had a mean SEN of 0.83, a mean SPE of 0.90, an AUC of 0.9290, and a Q* of 0.8640. The comparison between the diagnostic efficacy of SPECT and PET/CT showed that SPECT was superior for SEN (P = .0014) and PET/CT had a significantly better SPE (P = .001). The summary diagnostic efficacy between these modalities did not differ significantly (P > .05). CONCLUSIONS: The present clinical evidence showed that SPECT is an excellent tool to exclude patients with no mandibular invasion, but is not as good as PET/CT to confirm the diagnosis.

Fibular flap mandibular reconstruction for third-stage medication-related osteonecrosis of the jaw: A retrospective single-center study.

Background/purpose: The incidence and patient population of medication-related osteonecrosis of the jaw (MRONJ) has dramatically increased due to the use of drugs suppressing bone metastasis. However, its clinical treatment is still very difficult. The aim of this study was to evaluate the effectiveness and outcome of immediate fibular flap reconstruction for MRONJ in the mandible. Materials and methods: Patients who underwent immediate fibular flap reconstruction for MRONJ in the mandible in our institution from 1990 to 2022 were screened and identified. Their demographics, drug history, symptoms, surgical parameters and follow-up data were collected and analyzed. Results: In total, 25 patients with MRONJ stage 3 were included. The main cause of drug administration was osseous metastasis (88%), and zoledronate was the main drug. Pain, swelling (44%), pyorrhea (28%), extraoral fistulas (16%) and necrotic bone exposure (12%) were the main chief complaints. After segmental mandibulectomy, the fibular flap harvest was 9.73 ± 3.37 cm, and 18/25 (72%) were cut into two segments to reconstruct the mandible. Sixty-eight percent had an intraoral skin paddle placed. All of the flaps survived, and 21/25 (84%) of the soft tissue underwent primary healing. During follow-up, the alleviation of symptoms was effective, and there was no primary disease progression or death. Conclusion: This is the largest comprehensive investigation of fibular flap reconstruction for MRONJ in the mandible, which is proved to be an alternative and effective treatment for managing advanced patients with MRONJ.

Multiparametric CT-based assessment of pretracheal anatomical relationships for safe tracheotomy: A long-forgotten issue revisited.

OBJECTIVES: To assess the anatomical relationships and variations in the pretracheal space and to guide tracheotomy procedures in a safe manner with image-based evidence. MATERIALS AND METHODS: A retrospective study was conducted on unirradiated patients requiring elective tracheotomies. Preoperative contrast-enhanced CT (CECT)/CT venography (CTV) was applied for an anatomical evaluation of the pretracheal region. Vascular morphologies were compared for three vessels: the anterior jugular vein (AJV), the innominate artery (IA) and the inferior thyroid vascular plexus (ITVP). The relationships between the thyroid isthmus and the 2nd-4th tracheal rings were also analyzed. RESULTS: A total of 120 patients were identified, most of whom (n = 110, 91.7%) had head and neck squamous cell carcinomas. Patients with recognizable AJVs (n = 118) were divided into 3 groups: single-branch (n = 11, 9.2%), double-branch (n = 105, 87.5%), and multibranch (n = 2, 1.7%). In addition, IAs were categorized as low-bifurcation (n = 51, 42.5%), high-bifurcation (n = 40, 33.3%), platform (n = 27, 22.5%) and variant types (n = 2, 1.7%). Within the platform types, high-lying IAs (n = 15, 8.3%) might have interfered with the standard tracheal incisions due to possible IA-tracheal overlay. This interference was also related to the height of intraoperative tracheal incisions (rn = 0.364, P = 0.001). Within ITVPs, independent-trunk types were found in 71 cases (59.2%), while common-trunk types were found in 45 (37.5%). In addition, a low thyroid isthmus (suprasternal-isthmus distance <3 cm) was found in 83 cases (69.2%). CONCLUSIONS: CT image-based evidence can prepare junior practitioners with important pretracheal anatomical information, thereby facilitating safer tracheotomy procedures. Our results shed light on vascular relationships for emergent tracheotomy.

A comparative study on using superior thyroid artery perforator flaps versus traditional sternocleidomastoid myocutaneous flaps for reconstructions after oral cancer ablation: "New tricks for old dogs"?

OBJECTIVES: To introduce new superior thyroid artery perforator flaps (STAPF), and to compare the clinical outcomes with sternocleidomastoid myocutaneous flaps (SCMMF) for their intraoral applications. MATERIALS AND METHODS: Between January 2013 and December 2020, forty-three oral cancer patients who received post-oncologic reconstructions with one of these two regional flaps were retrospectively collected. Their techniques and outcomes were compared. All the STAPFs were preprepared with radiologic evaluations. RESULTS: Despite the common arterial origins, the compositions and harvesting procedures of STAPF and SCMMF were different. Though SCMMFs (n = 23) were designed in rotational styles, most STAPFs (n = 20) were septocutaneous perforator flaps, with 2 chimeric ones. In addition, the sizes of STAPFs were generally larger than those of SCMMFs (p = 0.006). Success rate for STAPFs was much higher, with only three partial cutaneous necroses. Radiotherapy delay was more frequently found in those reconstructed with SCMMFs (P = 0.046), mostly due to fistula formations. Besides, incomplete level IIB dissections were also reported in 9 (20.9%) patients in SCMMF group. In our study, the overall survival was affected by both flap conditions (p = 0.014, 1.333-12.881) and postoperative surgical complications (except fistula) (P = 0.005, 2.240-84.134). Functionally speaking, post-reconstructive speech and neck mobility (p < 0.001) were better in the STAPF group. CONCLUSIONS: With accumulated experiences on the use of locoregional flaps in the neck, STAPF, when well-prepared, can provide superior reconstructive outcomes for various intraoral defects. As a comparison with SCMMF in the same middle region, STAPF is a viable option with higher success rates and oncological safety for oral cancer patients.

Aberrant Methylation and Immune Microenvironment Are Associated With Overexpressed Fibronectin 1: A Diagnostic and Prognostic Target in Head and Neck Squamous Cell Carcinoma.

Background: Fibronectin 1 (FN1) is involved in cell adhesion and migration processes such as metastasis, wound healing, embryogenesis, blood coagulation, and host defense. However, the role of FN1 in the diagnosis and prognosis of head and neck squamous cell carcinoma (HNSCC) is far from understood. Methods: FN1 expression profiles and clinical parameters from multiple HNSCC datasets were applied to evaluate the association between FN1 expression and HNSCC survival. We also identified FN1 expression in the mRNA and protein levels in 20 pairs of clinical samples by quantitative polymerase chain reaction (qPCR) and immunohistochemistry. Receiver operator characteristic (ROC) analysis was used to demonstrate the potential diagnostic value of FN1 in HNSCC. Aberrant methylation PPI networks were established using multiple bioinformatic tools based on TCGA database. The immune microenvironment and levels of immune checkpoints were investigated between groups with high and low FN1 expression. Results: FN1 was significantly upregulated in HNSCC compared with para-carcinoma tissues on the basis of TCGA database and our clinical samples. Univariate and multivariate Cox regression analysis revealed that FN1 could be an independent indicator for prognosis of HNSCC. GO enrichment and KEGG pathway analysis demonstrated that cell adhesion, focal adhesion, and the PI3K-Akt signaling pathway might be involved in the potential mechanisms of FN1's prognostic performance in HNSCC. Methylation of FN1 was also higher and closely associated with poorer survival in HNSCC. In addition, FN1 expression was positively correlated with three DNA methyltransferases (DNMT1, DNMT3A, and DNMT3B). Furthermore, FN1 was positively associated with CD4+ T cells, endothelial cells, macrophages, and NK cells and negatively correlated with CD8+ T cells Conclusion: FN1 might be an independent prognostic biomarker for HNSCC patients. Hypermethylation, the aberrant proportions of immune cells, and the PI3K/Akt signaling pathway might be involved in the mechanism of FN1's oncogene role in HNSCC.

Autophagy is positively associated with the accumulation of myeloid­derived suppressor cells in 4­nitroquinoline­1­oxide­induced oral cancer.

It has previously been demonstrated that autophagy and inflammation act synergistically to promote carcinogenesis. However, the precise roles of autophagy in multistep oral carcinogenesis are still unclear, particularly regarding its association with tumor inflammation. The present study established a 4NQO­induced oral cancer mouse model and investigated autophagy status in the multistep process of oral carcinogenesis using immunohistochemistry, western blotting and immunofluorescence staining. Furthermore, the number of Gr­1+CD11b+ myeloid derived suppressor cells (MDSCs) and CD4+ Foxp3+ regulatory T cells (Tregs) during oral carcinogenesis and the association with autophagy status was also examined. The results revealed that the expression of autophagy biomarkers, including dihydrosphingosine 1-phosphate phosphatase LCB3 (LC3B), p62/SQSTM1 (p62) and Beclin 1 increased during 4NQO­induced carcinogenesis and in human oral cancer. The number of MDSCs and Tregs also increased during oral carcinogenesis. Furthermore, the expression of LC3B and p62 significantly correlated with the accumulation of MDSCs and the expression of Beclin 1 correlated with the increase of Tregs. These data indicated that autophagy may be activated by the tumor inflammation microenvironment during oral carcinogenesis.

LncRNAs as an intermediate in HPV16 promoting myeloid-derived suppressor cell recruitment of head and neck squamous cell carcinoma.

The emerging evidence showed that long noncoding RNAs (lncRNAs) are involved in cell growth and apoptosis as well as cancer progression and metastasis of malignant tumor, however, limited data are available on the role of lncRNAs in human papillomavirus (HPV)-associated Head and neck squamous cell carcinomas (HNSCC). Here, we demonstrated that 23.98% of 196 HNSCC cases in Southwest China could be classified as HPV16 infection. The number of MDSCs in HPV-positive HNSCC was significantly higher than normal control, indicating that HPV infection may promote MDSCs aggregation. Then, we applied an array-based approach to monitor the lncRNA expression between HPV-positive HNSCC, HPV-negative HNSCC and normal oral mucous, and obtained 132 different lncRNAs in different HPV infected states of HNSCC. HOTAIR, PROM1, CCAT1, and MUC19 mRNA levels, determined by qRT-PCR were inversely correlated with MDSCs collection of HPV-associated HNSCC in 2 independent patient cohorts. The results may provide a rationale for the further evaluation of lncRNAs as a molecular target to elucidate the molecular mechanism of HPV promoting MDSCs collection of HNSCC.

[Postoperative infection bacteria and drug resistance in patients with oral and maxillofacial tumors].

PURPOSE: To evaluate postoperative infection patterns of patients suffering from oral and maxillofacial neoplasms. The risk factors giving rise to postoperative infections were analyzed. Infection bacteria and antibiotic resistance were investigated. METHODS: Fifty-three cases suffering from postoperative infection were selected during 2007.12-2012.12 at the Department of Oral & Maxillofacial Surgery, West China College of Stomatology. The relationship between infections and factors including patients' sex, age, type of tumor, operation time and methods were evaluated with Excel and SPSS 21.0 software package, putting emphasis on infection bacteria and drug-resistance. RESULTS: Postoperative infection mainly occurred in patients with oral malignant tumors. Operation types and time had important influence on postoperative infection. The infection bacteria included gram-positive (59.5%) and gram-negative ones (40.5%). Streptococcus pyogenes accounted for the majority of G- bacteria, which was very sensitive to ß-lactam antibiotics. Pseudomonas aeruginosa were multi-drug resistant G- bacteria, which brought difficulties to the treatment of the infection. CONCLUSIONS: Integrant bacterial culture and drug sensitivity test should be performed to choose appropriate antibiotics, and monitor multi-drug resistant bacteria, so as to improve the control rates of postoperative infection.