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.

TP53-Associated Ion Channel Genes Serve as Prognostic Predictor and Therapeutic Targets in Head and Neck Squamous Cell Carcinoma.

TP53 mutations are the most occurred mutation in HNSCC which might affect the ion channel genes. We aim to investigate the ion channel gene alteration under TP53 mutation and their prognostic implication. The overall mutation status of HNSCC were explored. By screening the TP53-associated ion channel genes (TICGs), an ion channel prognostic signature (ICPS) was established through a series of machine learning algorithms. The ICPS was then evaluated and its clinical significance was explored. 82 TICGs differentially expressed between TP53WT and TP53MUT were screened. Using univariate regression analysis and LASSO regression analysis and multivariate regression analysis, an ICPS containing 7 ion channel genes was established. A series of evaluation was carried out which proved the predictive ability of ICPS. Functional analysis of ICPS revealed that cancer-related pathways were enriched in high-risk group. Next, for clinical application, a nomogram was constructed based on ICPS and other independent clinicopathological factors. TP53 mutation status strongly affects the expression of ion channel genes. The ICPM we have identified is a strong indicator for HNSCC prognosis and could help with patient stratification as well as identification of novel drug targets.

Magnesium promotes the viability and induces differentiation of neural stem cells both in vitro and in vivo.

OBJECTIVE: Neural stem cells (NSCs) are multipotent stem cells that generating various neural cells, including neurons, astrocytes and oligodendrocytes. This showed that NSCs is an ideal candidate in the application of neural disease treatment. In the current study, we established a simple and efficient method to promote the viability and induce the differentiation of NSCs by stimulating with magnesium. METHODS: The proliferation and differentiation of NSCs was determined by MTT assay and immunostaining. The behavior alteration was measured by rotorod test and Morris water maze. RESULTS: Magnesium enhanced proliferation in NSCs. The ratio of Nestin+, Ki67+ and GFAP+ progenitor cells was increased in the presence of magnesium. Besides, magnesium induced the glial differentiation instead of neuronal differentiation in NSCs. By contrast, transplantation of Mg2+-treated NSCs in vivo generated more neurons. In established PD models, transplantation of Mg2+-treated NSCs could improve the symptoms and recover the memory. CONCLUSION: We established a simple and efficient way to promote the proliferation and induce the differentiation of NSCs. More importantly, this may also facilitate to develop a new method to neural disorder treatment.

Long noncoding RNA GIHCG enhanced tongue squamous cell carcinoma progression through regulating miR-429.

Long noncoding RNAs play essential roles in cancer development and progression. Here, we tried to investigate the role of GIHCG in the progression and metastasis of tongue squamous cell carcinoma (TSCC). In our study, we showed that that the expression level of GIHCG was upregulated in TSCC tissues and cell lines. In addition, we indicated that high GIHCG expression was positively associated with poor overall survival. Moreover, ectopic expression of GIHCG enhanced TSCC cell cycle, proliferation, and migration. Elevated expression of GIHCG inhibited the miR-429 expression in TSCC cells. We demonstrated that the expression level of miR-429 was lower in TSCC tissues and cell lines. Low miR-429 expression was positively associated with poor overall survival. We then determined the correlation between miR-429 and GIHCG expression levels. A statistically significantly inverse correlation was observed between miR-429 and GIHCG expression levels in TSCC tissues. In addition, overexpression of miR-429 suppressed the TSCC cell cycle, proliferation, and migration. Elevated expression of GIHCG promoted TSCC cell cycle, proliferation, and migration through regulating miR-429 expression. These results suggested that GIHCG increased TSCC progression through negative modulation of miR-429. Our results suggested that GIHCG/miR-429 might play a vital role in TSCC progression.

Long non-coding RNA CASC15 promotes tongue squamous carcinoma progression through targeting miR-33a-5p.

Long non-coding RNAs (lncRNAs) have gained a lot of attention because they participate in several human disorders, including tumors. This study determined the role of LncRNA CASC15 (cancer susceptibility candidate 15) in the development of tongue squamous cell carcinoma (TSCC). Here, we identified that CASC15 expression was upregulated in TSCC samples and cell lines. We showed that overexpression of CASC15 promoted cell proliferation, cycle, and migration in TSCC. In addition, we revealed that miR-33a-5p expression was downregulated in TSCC tissues and cell lines. Moreover, we showed that the expression of CASC15 was negatively related with miR-33a-5p expression in TSCC tissues. Ectopic expression of miR-33a-5p suppressed cell proliferation, cycle, and migration in TSCC. Elevated expression of CASC15 suppressed miR-33a-5p expression and promoted ZEB1 expression in SCC4 cell. Ectopic expression of CASC15 promoted TSCC cell proliferation, cycle, and migration through targeting miR-33a-5p. These results suggested that lncRNA CASC15 and miR-33a-5p might be exploited as new markers of TSCC and were potential treatment targets for TSCC patients.

Cyclophilin a increases CD68+ cell infiltration in rat experimental periodontitis.

Cyclophilin A (CyPA) is a potent chemokine, which can directly induce leukocyte chemotaxis and contribute to the pathogenesis of inflammation-mediated diseases. This study is to observe the expression and distribution of CyPA and CD68+ cells in the histopathogenesis of rat ligation-induced experimental periodontitis, and assess the role of CyPA in CD68+ cell infiltration in rat experimental periodontitis. Experimental periodontitis was induced by ligation according to our previous method. CyPA expression in gingival tissues was detected by western blotting. Immunohistochemistry was applied for CyPA and CD68 distribution. For further certifying the role of CyPA in CD68+ cell infiltration, the right mandibular first molar received 0.1 µM CyPA locally by gingival injection every 2 days (L + C group), while the left mandibular first molar received saline as a control group (L group). The number of CD68+ cells in the experimental periodontitis was observed by immunohistochemistry. Alveolar bone destruction was assessed by micro-computerized tomography (micro-CT). Osteoclast was observed through TRAP staining. Nuclear factor (NF)-κB phospho-p65 (p p65) and phosphor-IκBα (p IκBα) expressions were detected to investigate NF-κB activation. CyPA showed an increasing trend at 1-6 weeks after ligation. CyPA and CD68+ cells were present in the gingival inflammatory infiltration, and participated in alveolar bone destruction. In the L + C group, the number of CD68+ cells was increased compared with the L group, and greater alveolar bone destruction was observed. NF-κB p p65 and p IκBα expressions were upregulated in the L + C group compared with the L group indicating NF-κB activation. CyPA increases CD68+ cell infiltration in rat experimental periodontitis, suggesting CyPA might be an anti-inflammatory therapeutic target.

EMMPRIN-CypA contributes to the inflammatory processes in human periodontitis through infiltrating CD68+ inflammatory cells.

Extracellular matrix metalloproteinase inducer (EMMPRIN) and its ligand cyclophilin A (CypA) levels increase in human inflammatory diseases, but EMMPRIN-CypA interactions and cell types expressing EMMPRIN and CypA in the pathogenesis of periodontitis are uncertain. Immunohistochemistry, immunofluorescence and western blotting revealed the level of EMMPRIN, CypA, and CD68 in human periodontitis. Double labelled immunofluorescence colocalized the expression of CD68 and CypA, and CD68 and EMMPRIN. Further investigation of EMMPRIN-CypA interactions and CD68+ infiltrating cells was applied using mouse monocyte cell line RAW264.7 in vitro. A higher level of EMMPRIN and CypA staining was detected in human periodontitis, compared with healthy gingiva. Many inflammatory cells, including CD68+ cells, infiltrated gingival tissues of human periodontitis. Both EMMPRIN and CypA could be localized in the CD68+ infiltrating cells. CypA could induce NF-κB activation by increasing expression of NFκB p-p65 in the nucleus of mouse monocytic cells RAW264.7 in vitro. EMMPRIN-CypA may contribute to the inflammatory processes in human periodontitis through infiltrating CD68+ inflammatory cells.

[Influence of smoking on apoptosis in human gingival epithelium].

PURPOSE: Cigarette smoking was a major risk factor for oral hygiene and periodontal status. The study was to investigate the relationship between different exposure to cigarette smoke and apoptosis in stratum spinosum and stratum basale cells of human gingival epithelium. METHODS: 47 smoking patients with crown lengthening surgery or extraction of the impacted teeth were enrolled. It comprised 41 males and 6 females. Their age ranged from 18 to 29 years old (average age: 21.4). According to smoke history (SH), average cigarettes per day (ACD) and total smoke number (TSM), 47 smokers were divided into group I (SH<5a or ACD<10 and TSM<15 thousand, n=19), group II (5a< or = SH or 10< or = ACD and 15 thousand < or =TSM, n=28). Another 10 non-smoking patients were chosen as control. Apoptosis was identified by the terminal deoxy-transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) method. Differences between the two groups were analyzed using Student-Newman-Keuls (SNK).The criteria for statistical significance was accepted at the probability level P<0.05. RESULTS: There was increased apoptosis in stratum spinosumin and stratum basale cells of smokers compared to that of non-smokers. Apoptosis in stratum spinosumin cells was significantly different between smokers and non-smokers (P<0.01), between group I and group II (P<0.01). CONCLUSION: It is conclude that exposure to cigarette smoke can increase apoptosis in stratum spinosumin cells of human gingival epithelium, which may influence its normal metabolism and protein secretion.