COL11A1 as a potential prognostic target for oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is a malignant tumor occurring in the oral cavity. However, the molecular mechanism of OSCC is not clear. Bioinformatics was used to screen and identify role of collagen type X1 alpha 1 (COL11A1) on OSCC. 200 patients with OSCC were recruited. Clinical and follow-up data were recorded and COL11A1 expression levels were tested. Pearson chi-square test and Spearman correlation coefficient were used to analyze relationship between prognosis and related parameters in patients with OSCC. Univariate and multivariate Logistic regression, univariate and multivariate Cox proportional risk regression were used for further analysis, survival curve was drawn. Through bioinformatics analysis, OSCC patients with higher expression of COL11A1 have poor overall survival compare with OSCC patients with lower expression of COL11A1 (hazard ratios [HR] = 1.32, P = .047). Pearson chi-square test showed that age (P = .011), tumor grade (P = .023), COL11A1 (P < .001) was significantly correlated with prognosis of OSCC. Univariate Logistic regression analysis showed age (odds ratio [OR] = 2.102, 95% confidence intervals [95%CI]: 1.180-3.746, P = .012), tumor grade (OR = 1.919, 95%CI: 1.093-3.372, P = .023) and COL11A1 (OR = 12.775, 95%CI: 6.509-25.071, P < .001). Multivariate Logistic regression analysis showed that COL11A1 (OR = 12.066, 95%CI: 6.042-24.096, P < .001) was significantly associated with prognosis of patients with OSCC. Univariate Cox regression analysis showed that age (HR = 1.592, 95%CI: 1.150-2.205, P = .005), tumor grade (HR = 1.460, 95%CI: 1.067-1.999, P = .018) and COL11A1 (HR = 1.848, 95%CI: 1.340-2.548, P < .001) were significantly correlated with survival time of OSCC patients. Multivariate Cox regression analysis showed that tumor grade (HR = 1.466, 95%CI: 1.064-2.020, P = .019) and COL11A1 (HR = 1.645, 95%CI: 1.164-2.325, P = .005) were significantly correlated with survival time of OSCC patients. COL11A1 is significantly correlated with occurrence of OSCC. When COL11A1 is highly expressed, prognosis of patients with OSCC is worse and the survival time is shorter.

Corneodesmosin as a potential target of oral squamous cell carcinoma.

OBJECTIVE: The relationship between oral squamous cell carcinoma (OSCC) and Corneodesmosin (CDSN) remains unclear. This study aims to explore the correlation between CDSN and the prognosis and survival time of patients with OSCC. METHODS: Bioinformatics were used to identify the hub role of CDSN in the OSCC. A total of 200 patients with OSCC were recruited. Clinical and follow-up data were recorded, and the expression level of CDSN was detected. Pearson chi-square test and Spearman correlation coefficient were used to analyze the relationship between prognosis and related parameters in patients with OSCC. Univariate and multivariate Logistic regression and Cox proportional risk regression were applied for further analysis, and receiver operating characteristic curve and survival curve of subjects were plotted. RESULTS: CDSN was identified as the most significant hub gene of the OSCC by the cytoHubba. By the comparative toxicogenomics database (CTD) analysis, there was strong relationship between the CDSN and mouth neoplasms, head and neck neoplasms, squamous cell carcinoma of head and neck. The OSCC patients with low expression level of CDSN have poor overall survival compared with the high expression level of CDSN (HR = 0.75, 95% confidence interval [95%CI]: 0.57-0.98, P = .036). Spearman correlation coefficient analysis showed that CDSN expression level was significantly correlated with prognosis (ρ = -0.528, P < .001). Multivariate Logistic regression analysis showed that poor prognosis (odds ratio [OR] = 0.096, 95%CI: 0.049-0.189, P < .001) was significantly associated with low expression of CDSN. Cox regression analysis showed that the survival time of OSCC patients was shorter when CDSN expression was low (HR = 0.588, 95%CI: 0.420-0.823, P = .002). Strong predictive value of CDSN for the OSCC survival time was obtained by the biological process (BP)-neural network and support vector machine (SVM). CONCLUSION: CDSN was significantly correlated with OSCC, and the shorter the survival time of patients with OSCC was, the worse the prognosis was.

Effect of pembrolizumab on T lymphocyte subsets in patients with advanced oral cancer and its therapeutic effect.

BACKGROUND: The aim of this study is to investigate changes of peripheral blood lymphocyte subsets before and after treatment with pembrolizumab for advanced oral cancer and its clinical sig-nificance. METHODS: 32 patients with advanced oral cancer who received pembrolizumab treatment were selected as observation group, 30 healthy people during the same period were selected as control group. Before treatment and in cycles 1, 2, 3 and 4 after treatment, fluid cytometry was used to detect changes in levels of lymphocyte subsets in peripheral blood of patients. RESULTS: CD3+, CD4+, CD4+/CD8 + indexes of patients with advanced oral cancer before treatment were significantly lower than those in control group (P < .05), CD8 + level was significantly increased (P < .05); After 1 cycle of pembrolizumab treatment, there was no significant difference in changes of lymphocyte subsets compared with before immunotherapy; After 2 and 3 cycles of treatment, CD3+, CD4+, CD4+/CD8 + values were higher than before the treatment (P > .05), CD8 + index was slightly lower than before treatment (P < .05); After fourth cycle of treatment, CD3+, CD4+, CD4+/CD8 + values were significantly improved compared to before treatment (P < .05), CD8 + index was significantly lower than before treatment (P < .05); In treatment process of patients with stable disease (SD)/partial response (PR), the CD3+, CD4+, CD4+/CD8 + values of fourth cycles were higher than before treatment (P < .05), CD8 + index was lower than before treatment (P < .05); During treatment of progressive disease (PD) patients, changes of lymphocyte subsets in fourth cycles were not significantly different from those before treatment (P > .05). This article shows through analysis that expression of programmed cell death ligand 1 (PD-L1) and pathological types have no obvious influence on effect of immunotherapy. Multi-factor analysis shows that it is more meaningful to observe the changes of CD3+, CD4 + and CD8 + at the same time to predict effect of immunotherapy. CONCLUSION: Pembrolizumab can regulate changes of T lymphocyte subsets in patients with advanced oral cancer, improve immune status of patients, there is no obvious adverse reaction. Monitoring changes of lymphocyte subsets during treatment can predict effect of immunotherapy.

MKI67 an potential oncogene of oral squamous cell carcinoma via the high throughput technology.

Oral squamous cell carcinoma is a malignant tumor that occurs in the oral cavity, with poor prognosis and easy recurrence. However, the relationship between MKI67 and oral squamous cell carcinoma remains unclear. The oral squamous cell carcinoma datasets GSE138206, GSE146483 and GSE184616 were downloaded from the gene expression omnibus database, and the differentially expressed genes (DEGs) were screened. The protein-protein interaction network was constructed and analyzed by search tool for the retrieval of interacting genes database and Cytoscape software. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) were used for functional enrichment analysis. GO and KEGG analyses were performed on the whole genome, as formulated by gene set enrichment analysis. comparative toxicogenomics database was used to identify the diseases most associated with the core genes. TargetScan was used to screen miRNA regulating central DEGs. A total of 1472 DEGs were identified. GO analysis showed that the differentially expressed genes were mainly enriched in the tissues of extracellular matrix, type i interferon signaling pathway, human papillomavirus infection, adhesion spot, hepatitis C and ECM-receptor interaction. Enrichment items were similar to GO and KEGG enrichment items of differentially expressed genes. 10 core genes were obtained, and their expression was different between oral squamous cell carcinoma and normal tissue samples. MKI67 is highly expressed in oral squamous cell carcinoma and may be an oncogene in oral squamous cell carcinoma.

SPP1 and FN1 are significant gene biomarkers of tongue squamous cell carcinoma.

Tongue squamous cell carcinoma (TSCC) is one of the most common malignant tumor types in the oral and maxillofacial region. The etiology and pathogenesis behind TSCC is complicated. In the present study, three gene expression profiles, namely GSE31056, GSE13601 and GSE78060, were downloaded from the Gene Expression Omnibus (GEO). The GEO2R online tool was utilized to identify differentially expressed genes (DEGs) between TSCC and normal tissue samples. Furthermore, a protein-protein interaction (PPI) network was constructed and hub genes were validated and analyzed. A total of 83 common DEGs were obtained in three datasets, including 48 upregulated and 35 downregulated genes. Pathway enrichment analysis indicated that DEGs were primarily enriched in cell adhesion, extracellular matrix (ECM) organization, and proteolysis. A total of 63 nodes and 218 edges were included in the PPI network. The top 11 candidate hub genes were acquired, namely plasminogen activator urokinase (PLAU), signal transducer and activator of transcription 1, C-X-C motif chemokine ligand 12, matrix metallopeptidase (MMP) 13, secreted phosphoprotein 1 (SPP1), periostin, MMP1, MMP3, fibronectin 1 (FN1), serpin family E member 1 and snail family transcriptional repressor 2. Overall, 83 DEGs and 11 hub genes were screened from TSCC and normal individuals using bioinformatics and microarray technology. These genes may be used as diagnostic and therapeutic biomarkers for TSCC. In addition, SPP1 and FNl were identified as potential biomarkers for the progression of TSCC.

Mechanism of Astragalus membranaceus in the treatment of laryngeal cancer based on gene co-expression network and molecular docking.

Astragalus membranaceus (HUANG QI, HQ) is a kind of traditional Chinese medicine. Researchers have widely concerned its antitumor effect. At present, there is still a lack of research on the treatment of laryngeal cancer with HQ. In this study, we integrated data from the weighted gene co-expression network of laryngeal cancer samples and the components and targets of HQ. A new method for dividing PPI network modules is proposed. Important targets of HQ treatment for laryngeal cancer were obtained through the screening of critical modules. These nodes performed differential expression analysis and survival analysis through external data sets. GSEA enrichment analysis reveals pathways for important targets participation. Finally, molecular docking screened active ingredients in HQ that could interact with important targets. Combined with the laryngeal cancer gene co expression network and HQ PPI network, we obtained the critical module related to laryngeal cancer. Among them, MMP1, MMP3, and MMP10 were chosen as important targets. External data sets demonstrate that their expression in tumor samples is significantly higher than in normal samples. The survival time of patients with high expression group was significantly shortened, which is a negative factor for prognosis. GSEA enrichment analysis found that they are mainly involved in tumor-related pathways such as ECM receptor interaction and Small cell lung cancer. The docking results show that the components that can well bind to important targets of HQ are quercetin, rutin, and Chlorogenic acid, which may be the primary mechanism of the anti-cancer effect of HQ. These findings provide a preliminary research basis for Chinese medicine treatment of laryngeal cancer and offer ideas to related drug design.

[Effect of miR-204-5p on the proliferation, migration, and invasion on tongue squamous cell carcinoma SCC25 cells by targeting bromodomain-containing protein 4].

OBJECTIVE: This study aimed to explore the target relationship between miR-204-5p and bromodomain-containing protein (BRD) 4, as well as their effects on cell proliferation, migration, and invasion in tongue squamous cell carcinoma SCC25. METHODS: Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to detect miR-204-5p and BRD4 expression levels in tongue squamous cell carcinoma and different cell lines. TargetScan and dual luciferase reporter assay were used to confirm the target relationship between miR-204-5p and BRD4. The effects of miR-204-5p on SCC25 cell proliferation were examined by cell counting kit (CCK) 8 assay, whereas those on SCC25 cell migration and invasion were determined by Transwell assay. RT-qPCR and Western blot were used to detect the effects of miR-204-5p mimics and inhibitors on BRD4 expression. Transwell and CCK8 assays were used to detect the effects of miR-204-5p on proliferation, migration, and invasion through BRD4 regulation. RESULTS: miR-204-5p was significantly downregulated in the tissues and cells of squamous cell carcinoma, and BRD4 showed the opposite result. The increase in miR-204-5p expression can inhibit the proliferation, migration, and invasion of SCC25 cells. TargetScan and luciferase test confirmed that miR-204-5p and BRD4 had a negative regulatory relationship with BRD4, respectively. Moreover, miR-204-5p mimics can inhibit BRD4 expression, and miR-204-5p inhibitors can promote BRD4 expression upregulation. When miR-204-5p and BRD4 were overexpressed in SCC25 cells, BRD4 can make up for the inhibitory effect of miR-204-5p on SCC25 cells. CONCLUSIONS: miR-204-5p could inhibit proliferation, migration and invasion in tongue squamous cell carcinoma SCC25 cells by targeting BRD4 gene.

[Effect of EDTA and hyaluronic acid on the activity of periodontal ligament fibroblasts and the expression of cytokines].

PURPOSE: To investigate the effect of ethylenediaminetetraacetic acid (EDTA) and hyaluronic acid (HA) on the activity of periodontal ligament fibroblasts (PDLFs) and the expression of cytokines. METHODS: Twelve wisdom teeth extracted due to orthodontics treatment were selected and prepared into 5 mm×4 mm root pieces, then divided into EDTA group, HA group, EDTA+HA group and untreated group according to different treatment methods. They were placed in the well plate and PDLFs inoculated on the root piece. After 24 and 48 h of inoculation, the cell proliferation of each group was detected by MTT assay. The adhesion of PDLFs was observed under microscope. The inflammatory cytokines (IL-6, IL-8 and IL-1ß and TNF-α) levels produced by PDLFs were determined by ELLISA and Western Blot. SPSS 22.0 software package was used for statistical analysis. RESULTS: After 24 h and 48 h inoculation, all the treatments promoted cell proliferation and adhesion of PDLFs compared with the untreated group, and the combined treatment promoted cell proliferation and adhesion significantly better than single treatment (P<0.05). The cell proliferation and adhesion effect of EDTA group and HA group increased with the inoculation time, without significant difference between the groups (P>0.05). The results of ELLISA and Western blot showed that compared with the untreated group, the treatment groups inhibited the expression of inflammatory factors IL-6, IL-1ß and TNF-α, and promoted the expression of IL-8, and the effect of EDTA+HA group was much more significant(P<0.05). CONCLUSIONS: EDTA+HA can significantly promote the growth of periodontal ligament fibroblasts and inhibit the expression of inflammatory cytokines, which may be related to its ability to enhance the adhesion of fibroblasts and then improve their viability.

[Expression and clinical significance of Spindly and Bub3 in oral squamous cell carcinoma].

PURPOSE: To investigate the expression and significance of Spindly and Bub3 in oral squamous cell carcinoma(OSCC). METHODS: Sixty-five patients with OSCC admitted to the Fourth Hospital of Hebei Medical University from March 2017 to March 2019 were enrolled. RT-PCR was used to detect the expression of Spindly and Bub3 mRNA in oral squamous cell carcinoma and adjacent normal tissues from the patients. OSCC cell line was cultured. After siRNA transfection interference with the expression of Spindly and Bub3 genes, cell viability was detected by MTT assay, and the cell migration ability was detected by scratch test. Statistical analysis was performed with SPSS 22.0 software package RESULTS: The expression levels of Spindly and Bub3 mRNA in OSCC were significantly higher in adjacent tissues(P<0.05). The expression of Spindly and Bub3 mRNA was related to TNM staging, clinical staging and lymph node metastasis (P<0.05). The 5-year survival rate of patients with high expression of Spindly, Bub3, Spindly/Bub3 were significantly higher than the counterparts with low expression, and the 5-year survival rate of patients with high expression of Spindly/Bub3 was significantly lower than that of patients with high expression of Spindly and Bub3(P<0.05). The expression level of Spindly in siRNA-Spindly group was significantly lower than that in Spindly negative control group and blank control group, and the expression level of Bub3 in siRNA-Bub3 group was also significantly lower than that in Bub3 negative control group and blank control group. The expression level of Spindly in siRNA-Spindly group was significantly lower than that in Spindly negative control group and blank control group, and the expression level of Bub3 in siRNA-Bub3 group was also lower than that in Bub3 negative control group and blank control group. The migration ability of cells in siRNA-Spindly group at 24 and 48 hours was significantly lower than that in Spindly negative control group and blank control group; the migration ability at 24 and 48 hours was significantly lower than that of Bub3 negative control group and blank control group(P<0.05). CONCLUSIONS: Spindly and Bub3 are highly expressed in OSCC. Specific inhibition of Spindly and Bub3 gene expression can reduce the proliferation and migration of cancer cells, which might be used as one of the targets for the treatment of OSCC.

Tetrandrine suppresses amyloid-ß-induced inflammatory cytokines by inhibiting NF-κB pathway in murine BV2 microglial cells.

Microglial cells play an important role in mediating neuroinflammation in Alzheimer's disease (AD) by production of a series of proinflammatory mediators and clearance of Aß peptides and senile plaques. Tetrandrine, a bisbenzylisoquinoline alkaloid isolated from the Chinese herb Radix Stephania tetrandra, has been demonstrated to decrease the expression of proinflammatory mediators by inhibition of NF-κB activation. Here we investigated whether tetrandrine may affect the phagocytosis of microglia and the expression of cytokines and NF-κB in murine BV2 microglial cells. We found that fibrillar Amyloid-ß (fAß) induced phagocytosis of microglia and dramatically increased the levels of interleukin 1 beta (IL-1ß) and tumor necrosis factor alpha (TNF-α) as well as the expression of phospho NF-κB p65 in microglia cultures. The treatment with tetrandrine resulted in downregulation of phospho NF-κB p65 expression and strikingly reduced the production of IL-1ß and TNF-α. However, tetrandrine did not affect fAß induced phagocytosis of microglia. In conclusion, tetrandrine can decrease microglial detriment of neurotoxicity while maintaining microglial benefit of neuroprotection. Tetrandrine may be an efficacious and promising remedy in the treatment of AD.

Tetrandrine attenuates spatial memory impairment and hippocampal neuroinflammation via inhibiting NF-κB activation in a rat model of Alzheimer's disease induced by amyloid-ß(1-42).

BACKGROUND: The neuroinflammation characterized by glial activation and release of proinflammatory mediators is considered to play a critical role in the pathogenesis of Alzheimer's disease (AD). Tetrandrine, a bisbenzylisoquinoline alkaloid isolated from the Chinese herb radix Stephania tetrandra, has been demonstrated to decrease the expression of proinflammatory mediators by inhibition of nuclear factor-κB (NF-κB) activation. The purpose of the study was to investigate effects of tetrandrine on experimental model of AD. MATERIALS AND METHODS: Tetrandrine was administered in a rat model of AD induced by amyloid-ß (Aß)(1-42). The learning and memory impairment was examined using Morris water maze; the extent of histological injury in hippocampus was determined by Nissl staining; NF-κB DNA binding activity was assessed by electrophoretic mobility shift assay; the expression of tumor necrosis factor (TNF)-α and interleukin (IL)-1ß was measured by enzyme-linked immunosorbent assay. RESULTS: A significant improvement was observed in learning and memory impairment in rats with tetrandrine, and the increase in NF-κB DNA binding activity, the over-expression in IL-1ß and TNF-α as well as the increased histological injury in hippocampus in rats induced by Aß(1-42) were significantly reduced following administration of tetrandrine. CONCLUSION: Tetrandrine could significantly ameliorate Aß(1-42)-induced spatial learning and memory impairment, and the beneficial effect of tetrandrine treatment could be linked, at least in part, to the inhibition of NF-κB activity and the downregulation of expression of IL-1ß and TNF-α, suggesting that administration of tetrandrine may provide a therapeutic approach for AD.

A modified protocol for the comet assay allowing the processing of multiple samples.

In the present study, we developed a modified protocol for the basic comet assay that increased efficiency without sacrificing assay reliability. A spreader was used to spread agarose-embedded cells on a slide, making the manipulation and processing of multiple samples easier. Using this technique, we are able to rapidly prepare five or more comet assay samples on one slide. To demonstrate the effect of the protocol modifications on assay reliability, we present an example of how the comet assay was used in our laboratory to analyze the effect of melatonin (N-acetyl-5-methoxitryptamine; MEL) on the DNA repair ability of Gentiana macrophylla Pall. protoplasts after irradiation with different doses of ultraviolet-B radiation. A slight, but statistically significant (P<0.01), dose-related protective effect of MEL was observed in our experiments. The first use of the comet assay was to confirm the antioxidant and DNA repair functions of MEL in plants. The modified protocol is cost-effective and provides substantial advantages over the conventional comet assay.

Glial-derived neurotrophic factor regulates intestinal epithelial barrier function and inflammation and is therapeutic for murine colitis.

Although enteric glial cells (EGCs) have been demonstrated to play a key role in maintaining intestinal epithelial barrier integrity, it is not known how EGCs regulate this integrity. We therefore hypothesized that glial-derived neurotrophic factor (GDNF) produced by EGCs might be involved in this regulation. Here we investigated the role of GDNF in regulating epithelial barrier function in vivo. Recombinant adenoviral vectors encoding GDNF (Ad-GDNF) were administered intracolonically in experimental colitis induced by dextran sulphate sodium (DSS). The disease activity index (DAI) and histological score were measured. Epithelial permeability was assayed using Evans blue dye. The anti-apoptotic potency of GDNF in vivo was evaluated. The expression of tumour necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and myeloperoxidase (MPO) activity were measured by ELISA assay and/or RT-PCR. The expression of ZO-1, Akt, caspase-3, and NF-kappaB p65 was analysed by western blot assay. Our results showed that GDNF resulted in a significant reduction in enhanced permeability, inhibited MPO activity, IL-1beta and TNF-alpha expression, and increased ZO-1 and Akt expression. Moreover, GDNF strongly prevented apoptosis in vivo and significantly ameliorated experimental colitis. Our findings indicate that GDNF participates directly in restoring epithelial barrier function in vivo via reduction of increased epithelial permeability and inhibition of mucosal inflammatory response, and is efficacious in DSS-induced colitis. These findings support the notion that EGCs are able to regulate intestinal epithelial barrier integrity indirectly via their release of GDNF in vivo. GDNF is namely an important mediator of the cross-talk between EGCs and mucosal epithelial cells. GDNF may be a useful therapeutic approach to the treatment of inflammatory bowel disease.

Amelioration of dextran sulfate sodium-induced colitis by neuropeptide Y antisense oligodeoxynucleotide.

BACKGROUND: Neuropeptide Y (NPY) from enteric neurons has been shown to play an important role in immune and inflammatory responses. The purpose of the present study was to investigate the effects of NPY antisense oligodeoxynucleotides (ODNs) on an experimental model of ulcerative colitis (UC). METHODS: NPY antisense ODNs were administered in experimental colitis induced by dextran sulfate sodium (DSS). The disease activity index (DAI) and histological score were observed. The tumor necrosis factor (TNF)-alpha and NPY levels were measured by enzyme-linked immunosorbent assay. Phosphorylated Akt (p-Akt) expression was determined by immunohistochemical staining. Activated nuclear factor (NF)-kappaB was assessed by western blot analysis. Myeloperoxidase (MPO) activity was determined by using MPO assay kit. RESULTS: A significant improvement was observed in DAI and histological score in rats with NPY antisense ODNs, and the increase in NPY and TNF-alpha levels, MPO activity, and the expression p-Akt and p-NF-kappaB in rats with DSS-induced colitis was significantly reduced following the administration of NPY antisense ODNs. CONCLUSION: The administration of NPY antisense ODNs leads to an amelioration of DSS-induced colitis, suggesting that NPY plays an important role in modulating inflammation in colitis, and NPY antisense ODNs may be a useful therapeutic approach to the treatment of UC.