Alcohol consumption, which affects the structure and composition of the laryngeal microbiota, is one of the most important risk factors for laryngeal squamous cell cancer (LSCC). Our results demonstrated that high enrichment of Fusobacterium nucleatum (F. nucleatum) in LSCC was associated with poor prognosis. F. nucleatum increased miR-155-5p and miR-205-5p expression to suppress alcohol dehydrogenase 1B (ADH1B) and transforming growth factor β receptor 2 (TGFBR2) expression by activating innate immune signaling, resulting in ethanol metabolism reprogramming to allow F. nucleatum accumulation and PI3K/AKT signaling pathway activation to promote epithelial-mesenchymal transition, further exacerbating the uncontrolled progression and metastasis of LSCC. Therefore, the positive feed-forward loop between F. nucleatum and ethanol metabolism reprogramming promotes cell proliferation, migration, and invasion to affect LSCC patient prognosis. The amount of F. nucleatum is a potential prognostic biomarker, which yields valuable insight into clinical management that may improve the oncologic outcome of patients with LSCC.

Microbial community structure and its shift after laryngectomy in laryngeal squamous cell carcinoma (LSCC) patients were investigated. Thirty swab samples of LSCC prior to laryngectomy (SLC), 18 samples after 1-weeks laryngectomy (SLCA1w), and 30 samples after 24-week laryngectomy (SLCA24w) from 30 LSCC patients were sourced. Microbial diversity was profiled through sequencing the V3-4 variable region of the 16S ribosomal ribonucleic acid (rRNA) gene. Quantitative real-time polymerase chain reaction (qPCR) was used to validate the V3-4 region of 16S rRNA sequence data. The community structure and function of throat microbiota were assessed by PICRUSt analysis. Both Alpha and Beta diversity results showed significant differences in the throat microbiota of LSCC patients before and after laryngectomy (P < 0.05). Drinking index of SLC group was positively associated with the genus abundance of Prevotella (P < 0.05). SLCA1w group had lower abundance of Fusobacterium, Leptotrichia, Lachnoanaerobaculum and Veillonella compared to the SLC group (P < 0.05). SLCA24w group had higher abundance of Streptococcus and Leptotrichia, as well as the lower abundance of Fusobacterium and Alloprevotella compared to the SLC group (P < 0.05). The different propensity of throat microbiome of SLC group could implicate human cancer signaling pathways evident through Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) analysis (P < 0.05). Our study clarifies altered throat microbial community structure and function in LSCC patients during the perioperative period and post-operative recovery period. Importance Laryngeal squamous cell carcinoma greatly impacts patient9s livelihood, and non-invasive ways for prognostic assessment is valuable in determining the effectiveness of laryngectomy. We set to study the microbial structure changes in throat before and after laryngectomy and found the gene functions of several throat bacteria to be associated with human cancer signaling pathways. Our findings may offer insights into the disease management of patients with laryngeal squamous cell carcinoma. We hope to provide a perspective in molecular mechanisms to improve the prognosis of laryngeal cancer treatment and to facilitate relevant research.

Background: To investigate how Fusobacterium nucleatum (Fn) promotes oxidative stress and mediates proliferation and autophagy in hypopharyngeal squamous cell carcinoma (HPSCC). Methods: The prognosis for 82 HPSCC cases was retrospectively analyzed. HPSCC cell line FaDu was co-cultured with Fn. Knockdown of NUDT1 (shNUDT1 group) was done after observing DNA damage response. CCK8 and tumorigenesis assays for proliferation observation, mitochondria ROS (MitoROS) measurement to examine intracellular oxidative stress, and ELISA to analyze concentration of 8-oxo-2’-deoxyguanosine (8-oxo-dG) in cells. Dual-luciferase reporter assays clarified miR-361-3p connection with NUDT1. Autophagy flow was observed using electron microscopy and related proteins. Results: Fn was highly associated with NUDT1. The shNUDT1 group experienced lower proliferation compared with normal FaDu (NC group) in vivo and in vitro. The shNUDT1 group showed 8-oxo-dG and γH2AX to be elevated. Intracellular ROS decreased in shNUDT1Fn group when compared to Fn group. Upregulating miR-361-3p could suppress NUDT1 expression and downstream proliferation and autophagy. Fn modulated miR-361-3p via OH⁻, which could be proven by H₂O₂ assay and N-acetylcysteine. Conclusions: Higher Fn in HPSCC patients suggests poorer prognosis. NUDT1 might affect cell proliferation and autophagy and modulate DNA damage response. The oxidative stress induced miR-361-3p/NUDT1 axis is first introduced in microbiome-carcinoma research.

Objective: High levels of red cell distribution width (RDW) may be associated with adverse outcomes in patients with cancer. The purpose of the present study was to investigate the prognostic impact of pretreatment RDW levels on overall survival (OS), cancer-specific survival (CSS), and disease-free survival (DFS) in a large cohort of male laryngeal squamous cell cancer (LSCC) patients. Methods: A total of 809 LSCC patients who were treated between 2007 and 2011 at the Eye & ENT Hospital of Fudan University were enrolled and evaluated retrospectively. OS, CSS, and DFS were analyzed using the Kaplan–Meier method. To evaluate the prognostic significance of RDW levels, univariate, and multivariate Cox analyses were applied. Results: Higher pretreatment RDW levels were significantly associated with high death events, red blood cell count, hemoglobin, radiotherapy, operation therapy, and advanced tumor stage (p < 0.05). From the univariate analysis, we observed that the higher (13.2–13.5%) and the highest (>13.5%) quartiles of RDW level were consistent factors for poor OS, CSS, and DFS in LSCC patients. In the multivariate analysis, after adjusting for confounding factors, the higher and highest quartiles of RDW levels were identified as independent prognostic factors in male LSCC patients. Conclusion: Higher pretreatment RDW levels were demonstrated to be associated with poor clinical outcome in male LSCC patients and might be novel markers for patient stratification in LSCC management.