Identification of Immune Subtypes for Predicting the Prognosis of Patients in Head and Neck Squamous Cell Carcinoma.

Head and neck squamous cell carcinoma (HNSCC) is a common malignancy with poor prognosis and immune response, which plays an important role in tumor progression. Recently, immunotherapies have revolutionized the therapeutic means of malignancies including HNSCC. However, the relationship between immunophenotypes of HNSCC and its clinical response to immune-checkpoint inhibitors remains unclear. We aim to identify molecular subtyping related to distinct immunophenotypes in HNSCC. Consensus clustering algorithm was conducted for subtyping. Immunophenotypes between subtypes were compared according to infiltrating immunocytes, immune reactions, major histocompatibility complex (MHC) family, immunoinhibitory, immunostimulatory and immune scores. The relationship between immunophenotype and genotype was investigated from gene mutation and tumor mutation burden. The potential response of Immune-checkpoint blockade (ICB) therapy was estimated with TIDE and ImmuCellAI algorithms, and immune-checkpoint genes. The immune characteristics were also investigated. Biological functions were annotated by the gene-set enrichment analysis (GSEA) algorithm. Two distinct immune subtypes of HNSCC with different survival outcomes, biological characteristics, immunophenotype, and ICB response were identified. The subtype-1 was featured with better prognosis, more infiltrated immunocytes, stronger immune reaction, higher immune-related gene expression, higher immune-checkpoint gene expression (PD-1, PD-L1, and CTLA-4), and better ICB response. A higher immune response in subtype-1 was also revealed by GSEA. Subtype-1 possessed a higher immune response and more sensitivity to ICB therapy leading to a better prognosis. These findings may shed promising light on the immunotherapy strategy in HNSCC.

CKS2 modulates cell-cycle progression of tongue squamous cell carcinoma cells partly via modulating the cellular distribution of DUTPase.

BACKGROUND: CKS2 (CDC28 Protein Kinase Regulatory Subunit 2) is a gene that encodes CKS2 protein that has been characterized as a binding partner of the catalytic subunit of the cyclin-dependent kinases. However, its expression profile and regulatory effects in tongue squamous cell carcinoma has not yet been explored. METHODS: Bioinformatic analysis was conducted using bulk-seq data from The Cancer Genome Atlas and single-cell RNA-seq data from GSE103322. SCC9 and CAL27 cells were used as in vitro cell models for cellular and molecular studies. RESULTS: CKS2 expression was significantly upregulated in tongue squamous cell carcinoma tissues (N = 128) compared with adjacent normal tissues (N = 13). Its upregulation was associated with significantly shorter disease-specific survival and progression-free survival. Cellular status estimation in tumor cells indicated that CKS2 expression was moderately and positively correlated with cell-cycle progression. CKS2 inhibition in SCC9 and CAL27 cells resulted in decreased proliferation, weakened colony formation capability, and cell-cycle arrest at the G2/M phase. Immunofluorescence staining and co-Immunoprecipitation (co-IP) assay confirmed co-localization and interaction between CKS2 and DUTPase. CKS2 knockdown did not alter DUTPase expression but reduced its nuclear distribution. Both CKS2 and DUT expression were moderately correlated with their gene-level copy number. CONCLUSION: CKS2 expression is associated with unfavorable survival of patients with tongue squamous cell carcinoma. Inhibiting its expression could reduce tongue squamous cell carcinoma cell growth and induce G2/M arrest. CKS2 may interact with DUTPase and regulate its nuclear localization. Gene-level copy amplification might be an important mechanism of upregulated CKS2 and DUT in the tumor.

Serum Annexin A2 Level Is Associated With Diagnosis and Prognosis in Patients With Oral Squamous Cell Carcinoma.

PURPOSE: In several human cancer types, serum annexin A2 levels are increased, but little is known regarding oral squamous cell carcinoma (OSCC). This study aimed to measure serum annexin A2 levels in OSCC patients and assess the association with diagnosis and prognosis. MATERIALS AND METHODS: This case-control study compared serum annexin A2 concentrations in a group of OSCC patients and a control group. The predictor variable was the presence or absence of OSCC, and the outcome variable was the level of serum annexin A2. Annexin A2 concentrations were measured with an enzyme-linked immunosorbent assay, and correlations with clinicopathologic characteristics of OSCC were further evaluated. Receiver operating characteristic (ROC) curves, Kaplan-Meier curves, log-rank analyses, and a Cox proportional hazards model were used to evaluate the diagnostic and prognostic value of annexin A2. RESULTS: Serum samples were taken from 399 individuals: 126 patients with OSCC (aged 62.7 ± 10.6 years, 79 men and 47 women); 115 patients with benign oral disease (aged 63.9 ± 10.8 years, 73 men and 42 women); and 158 healthy controls (aged 65.4 ± 12.8 years, 92 men and 66 women). The annexin A2 level was significantly higher in OSCC patients than in patients with benign disease and controls (27.1 ± 9.81 ng/mL vs 15.9 ± 6.97 ng/mL and 15.0 ± 6.69 ng/mL, respectively). To distinguish OSCC patients from the other 2 groups, ROC curve-area under the ROC curve (AUC) analysis for serum annexin A2 levels provided an AUC of 0.80 (sensitivity, 0.62; specificity, 0.87) and an AUC of 0.77 (sensitivity, 0.57; specificity, 0.89). Furthermore, OSCC patients with high annexin A2 levels had poorer overall survival. CONCLUSIONS: This study suggested that an elevated serum annexin A2 level might be a novel diagnostic and prognostic biomarker for OSCC patients.

Upregulated effects of miR-7 in methicillin-resistant Staphylococcus aureus.

The aim of the study was to investigate the characteristic function of the upregulated effects of miR-7 in methicillin-resistant Staphylococcus aureus (MRSA). After separating the MRSA in clinic, the expression of miR-7 mRNA was tested by reverse transcription polymerase chain reaction. The overexpression, inhibition of miR-7, and control group were established by plasmid in vitro. Following transfection of the bacterial strain, the effect of ß-lactam antibiotics in minimum inhibitory concentration (MIC) was observed using the microporous dilution method, and antibacterial effects in vitro were observed using the dynamic growth curve method. The expression of miR-7 in sensitive MRSA was upregulated distinctly, with significant difference (P<0.05). MIC and the number of bacteria in the miR-7 overexpression group significantly increased while the inhibition group decreased prominently, with significant difference (P<0.05). The control and null plasmid groups revealed no significant difference. In conclusion, miR-7 upregulated the antimicrobial activity of MRSA, and the intervention of its expression may become a possible antibacterial target.

Dual targeting biomimetic liposomes for paclitaxel/DNA combination cancer treatment.

Combinations of chemotherapeutic drugs with nucleic acid has shown great promise in cancer therapy. In the present study, paclitaxel (PTX) and DNA were co-loaded in the hyaluronic acid (HA) and folate (FA)-modified liposomes (HA/FA/PPD), to obtain the dual targeting biomimetic nanovector. The prepared HA/FA/PPD exhibited nanosized structure and narrow size distributions (247.4 ± 4.2 nm) with appropriate negative charge of -25.40 ± 2.7 mV. HA/FA/PD (PTX free HA/FA/PPD) showed almost no toxicity on murine malignant melanoma cell line (B16) and human hepatocellular carcinoma cell line (HepG2) (higher than 80% cell viability), demonstrating the safety of the blank nanovector. In comparison with the FA-modified PTX/DNA co-loaded liposomes (FA/PPD), HA/FA/PPD showed significant superiority in protecting the nanoparticles from aggregation in the presence of plasma and degradation by DNase I. Moreover, HA/FA/PPD could also significantly improve the transfection efficiency and cellular internalization rates on B16 cells comparing to that of FA/PPD (p < 0.05) and PPD (p < 0.01), demonstrating the great advantages of dual targeting properties. Furthermore, fluorescence microscope and flow cytometry results showed that PTX and DNA could be effectively co-delivered into the same tumor cell via HA/FA/PPD, contributing to PTX/DNA combination cancer treatment. In conclusion, the obtained HA/FA/PPD in the study could effectively target tumor cells, enhance transfection efficiency and subsequently achieve the co-delivery of PTX and DNA, displaying great potential for optimal combination therapy.