Identification of potential therapeutic target genes,key miRNAs and mechanisms in oral lichen planus by bioinformatics analysis

The study aimed to identify the potential target genes and key miRNAs as well as to explore the underlying mechanisms in the pathogenesis of oral lichen planus (OLP) by bioinformatics analysis. The microarray data of GSE38617 were downloaded from Gene Expression Omnibus (GEO) database. A total of 7 OLP and 7 normal samples were used to identify the differentially expressed genes (DEGs) and miRNAs. The DEGs were then performed functional enrichment analyses. Furthermore, DEG-miRNA network and miRNA-function network were constructed by Cytoscape software. Total 1758 DEGs (598 up- and 1160 down-regulated genes) and 40 miRNAs (17 up- and 23 down-regulated miRNAs) were selected. The up-regulated genes were related to nuclear factor-Kappa B (NF-κB) signaling pathway, while down-regulated genes were mainly enriched in the function of ribosome. Tumor necrosis factor (TNF), caspase recruitment domain family, member 11 (CARD11) and mitochondrial ribosomal protein (MRP) genes were identified in these functions. In addition, miR-302 was a hub node in DEG-miRNA network and regulated cyclin D1 (CCND1). MiR-548a-2 was the key miRNA in miRNA-function network by regulating multiple functions including ribosomal function. The NF-κB signaling pathway and ribosome function may be the pathogenic mechanisms of OLP. The genes such as TNF, CARD11, MRP genes and CCND1 may be potential therapeutic target genes in OLP. MiR-548a-2 and miR-302 may play important roles in OLP development.     

牙周炎组织中miRNA表达谱差异筛选及功能预测分析

目的 筛选牙周炎患者组织中的差异表达miRNA,探讨其生物学功能以及参与的信号通路。方法 通过对微阵列数据库GSE54710中的158例牙周炎患者和40例健康人的牙龈组织中的基因芯片数据进行生物信息学分析,筛选差异表达miRNA,并预测参与的生物学功能和信号通路。采用SPSS 19.0软件包对数据进行统计学分析。结果 5种miRNAs（hsa-miR-451、hsa-miR-223、hsa-miR-486-5p、hsa-miR-3917、hsa-miR-671-5p）显著上调,4种miRNAs（hsa-miR-203、hsa-miR-210、hsa-miR-1246、hsa-miR-1260 ）显著下调。其中,hsa-miR-1260的靶基因584个,hsa-miR-451的靶基因139个。KEGG通路富集分析显示,hsa -miR-1260靶基因显著富集到TGF-beta等12条信号通路,hsa-miR-451靶基因显著富集到17条信号通路。结论 得到牙周炎组织中miRNAs的表达谱,牙周炎诱导的hsa-miR-1260和hsa-miR-451可能在牙周炎的生理病理学中起到关键作用。     

MicroRNA-203 up-regulates nitric oxide expression in temporomandibular joint chondrocytes via targeting TRPV4

ObjectiveMicroRNAs (miRNAs) are recognised as important regulators of a variety of fundamental biologic processes. Our study was undertaken to examine the role of MicroRNA-203 (miR-203) in modulating nitric oxide (NO) expression in female Sprague–Dawley rat mandibular condylar chondrocytes (MCCs) via targeting transient receptor potential vanilloid 4 (TRPV4) and to demonstrate the possible mechanism of NO inhibition by chondroprotective factor 17β-oestradiol (E2).MethodsThe expression of TRPV4 in mandibular condylar cartilage tissue and MCCs was detected by immunohistochemistry, immunofluorescence (IF), RT-PCR and Western blot, respectively. Primary SD rat MCCs were exposed to lipopolysaccharide (LPS), plus Ruthenium Red, 4α-phorbol 12,13-didecanoate (4αPDD), over-expressed miR-203 or E2 (10^?9 to 10^?6 M), the cellular supernatants were used for NO assay, miR-203 levels were measured by quantitative RT-PCR while TRPV4 expression changes were analysed by Western blot. The dual luciferase activity assay was performed to identify the target gene of miR-203.ResultsTRPV4 and miR-203 were stably expressed in MCCs. The MCCs’ expression of NO evoked by LPS could be enhanced or depressed by Ruthenium Red or 4αPDD. The dual luciferase assay suggested that TRPV4 was the direct target gene of miR-203. Over-expression of miR-203 inhibited the expression of TRPV4 and increased NO expression in MCCs. E2 inhibited NO expression by inhibition of miR-203, which was concurrent with the up-regulation of TRPV4 expression level in MCCs.ConclusionOur findings first suggested that miR-203 could up-regulate NO expression in female rat MCCs via targeting TRPV4. Moreover, the inhibition of NO by E2 might be at least in part through this mechanism.     

Urine metabolic profiling for the pathogenesis research of erosive oral lichen planus

ObjectiveOral lichen planus (OLP) is a relatively common chronic immune-pathological and inflammatory disease and potentially oral precancerous lesion. Erosive OLP patients show the higher rate of malignant transformation than patients with non-erosive OLP. Identifying the potential biomarkers related to erosive OLP may help to understand the pathogenesis of the diseases.MethodsMetabolic profiles were compared in control and patient subjects with erosive OLP by using ultra-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UPLC-QTOF-MS) coupled with pattern recognition methods An integrative analysis was used to identify the perturbed metabolic pathways and pathological processes that may be associated with the disease.ResultsIn total, 12 modulated metabolites were identified and considered as the potential biomarkers of erosive OLP. Multiple metabolic pathways and pathological processes were involved in erosive OLP.ConclusionThe dysregulations of these metabolites could be used to explain the pathogenesis of the disease, which could also be the potential therapeutic targets for the disease.     

口腔鳞状细胞癌预后相关基因标志物的分析

［摘要］ 目的 通过生物信息分析途径对口腔鳞状细胞癌患者与正常人群的差异基因进行分析,从分子水平探讨关键基因以及参与的信号通路,初步探索口腔鳞状细胞癌发生、发展的基因标志物。方法 从公共数据库基因表达数据库（GEO）中下载口腔鳞状细胞癌的相关芯片数据（GSE3524和GSE6631）,筛选出口腔鳞状细胞癌组织和对照组织表达有显著差异的基因。并对其功能及预后进行分析。结果 共筛选出129个差异表达基因,其中表达上调45个,下调84个,对其进行基因本体、京都基因与基因组百科全书和蛋白互作网络分析,发现分泌型焦磷酸蛋白（secreted phosphoprotein 1, SPP1）、金属基质蛋白酶（matrix metalloproteinase 1,MMP1）、丝氨酸蛋白酶抑制剂（serpin family E member 1,SERPINE1）、纤溶酶原激活剂（plasminogen activator urokinase,PLAU）处于基因核心节点位置。同时,根据这些关键基因表达水平的高低对口腔鳞状细胞癌患者进行分组,高表达组患者生存时间均低于低表达组,差异有统计学意义（PSPP1=0.045、PMMP1=0.046、PSERPINE1=0.0024和PPLAU=0.00049）。结论 基因组学分析方法筛选出的关键基因和信号通路有助于研究口腔鳞状细胞癌发生、发展的机制,也进一步为治疗靶点及预后分子标志物的选择提供了依据。     

Altered microRNA expression profile with miR-27b down-regulation correlated with disease activity of oral lichen planus

Oral Diseases (2012) 18 , 265–270 Background: Increasing evidence indicates that microRNAs (miRNAs) play a vital role in the pathogenesis of inflammatory and autoimmune diseases. The objective of this study was to investigate the altered miRNA expression profile in patients with oral lichen planus (OLP) and determine the miR‐27b expression. Methods: We compared miRNA expression patterns in oral biopsy specimens from patients with OLP (n = 3) with those from normal controls (n = 3) using microarray technology. We further assessed the miR‐27b expression in specimens from patients with OLP (n = 53) against controls (n = 34) using real‐time quantitative PCR (RT‐QPCR), and miR‐27b expression in specimens from patients with OLP (n = 15) against controls (n = 12) using in situ hybridization (ISH). Results: Using microarray analysis, a total of 46 differentially expressed miRNAs with more than 2‐fold change were identified, including 8 up‐regulated and 38 down‐regulated miRNAs. Both RT‐QPCR and ISH analyses revealed that miR‐27b was significantly down‐regulated in OLP tissue, and miR‐27b expression was even more suppressed in atrophic‐erosive OLP than in reticular OLP. In addition, miR‐27b was found to be expressed in the epithelial keratinocyte layer of both normal and OLP tissues. Conclusion: These data indicate that miRNAs may be the novel candidate biomarkers for the implication of miRNAs in the pathogenesis of OLP.     

Effect of Myd88 deficiency on gene expression profiling in salivary glands of female non-obese diabetic (NOD) mice

ObjectivesSjögren's syndrome (SS) is a chronic autoimmune disease characterized by inflammatory lesions in the salivary and lacrimal glands, which are caused by distinct lymphocytic infiltrates. Female non-obese diabetic (NOD) mice spontaneously develop inflammatory lesions of the salivary glands with SS-like pathological features. Previous studies have shown that MyD88, a crucial adaptor protein that activates innate immune signaling, affects lymphocytic infiltration, but its detailed role remains unclear. In this study, we investigated the role of MyD88 through gene expression profiling in the early phase of pathogenesis in the salivary glands of female NOD mice.MethodsSubmandibular glands collected from 10-week-old female wild-type and Myd88-deficient NOD mice were used for RNA preparation, followed by microarray analysis. The microarray dataset was analyzed to identify Myd88-dependent differentially expressed genes (DEGs). Data generated were used for GO enrichment, KEGG pathway, STRING database, and INTERFEROME database analyses.ResultsMyd88 deficiency was found to affect 230 DEGs, including SS-associated genes, such as Cxcl9 and Bpifa2. Most of the DEGs were identified as being involved in immunological processes. KEGG pathway analysis indicated that the DEGs were putatively involved in autoimmune diseases, such as systemic lupus erythematosus and rheumatoid arthritis. Furthermore, the DEGs included 149 interferon (IFN)-regulated genes.ConclusionsMyD88 is involved in the expression of specific genes associated with IFN-associated immunopathological processes in the salivary glands of NOD mice. Our findings are important for understanding the role of MyD88-dependent innate immune signaling in SS manifestation.     

Blood-based circulating microRNAs as potential biomarkers for predicting the prognosis of head and neck cancer—a systematic review

Objective

The aim of the present study was to systematically review the role of circulating miRNAs as potential prognostic biomarkers in head and neck cancer patients.

Materials and methods

PubMed, EMBASE, Scopus, Web of Science, and gray literature from January 1990 up to and including September 2019 were searched. The study selection was performed by two independent reviewers according to eligibility criteria.

Results

A total of 13 studies that met the eligibility criteria were included. Significant number of studies were executed majorly in China and predominant number of them were case-control in nature. A total of 22 different miRNAs were found to be concomitant with very poor prognosis in cancers of the head and neck region. Of these, eighteen miRNAs (miR-375, miR-1234, miR-103, miR-638, miR-200b-3p, miR-191-5p, miR-24-3p, miR-572, miR-483-5p, miR-20a, miR-22, miR-29a, miR-29b, mir-let-7c, miR-17, miR-374b-5p, miR-425-5p, and miR-196a) were upregulated and four miRNAs (miR-9, miR-29c, miR-223, and miR-187∗) were downregulated. The hazard ratio (HR) ranged from twofold to fivefold.

Conclusion

Based on the results, circulating miRNA may assist in the prediction of prognosis of head and neck cancer. Further multi-center randomized controlled clinical trials with large sample size are required to validate the results of the present review.

Clinical relevance

Decoding the circulating miRNA profile could aid in accurate prognostication of head and neck cancer.

     

Poor Prognosis of Oral Squamous Cell Carcinoma Correlates With ITGA6

ObjectivesOral cancer is the ninth most common cancer worldwide and a leading cause of cancer-related death. Oral squamous cell carcinoma (OSCC) accounts for 90% of all oral cancers. Autophagy is a conserved essential catabolic process related to OSCC. The aim of this study was to elucidate diagnostic and prognostic autophagy-related biomarkers in OSCC.MethodsThe OSCC gene expression data set was obtained from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) between the OSCC samples and adjacent healthy tissues were identified by R software. The Human Autophagy Database was screened, which revealed 222 autophagy-related genes. The autophagy-related DEGs were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were applied. Protein–protein interaction network analysis was performed in the STRING database. cytoHubba in the Cytoscape software was applied to determine the top 10 hub genes. The data set of patients with OSCC from The Cancer Genome Atlas (TCGA) was used to evaluate the prognostic value of the 10 hub genes. The association between prognosis-related hub genes and immune infiltrates was explored.ResultsTwenty-seven autophagy-related DEGs were identified. The top 10 hub genes were CCL2, CDKN2A, CTSB, CTSD, CXCR4, ITGA6, MAP1LC3A, MAPK3, PARP1, and RAB11A. ITGA6 was identified as the most efficient biomarker. Receiver operating characteristic curve analysis indicated that ITGA6 had the highest diagnostic accuracy for OSCC (area under the curve = 0.925). ITGA6 expression was significantly related to immune infiltrates.ConclusionsThe autophagy-related gene ITGA6 might be an efficient diagnostic and prognostic biomarker in OSCC.     

MicroRNA expression correlates with disease recurrence and overall survival in oral squamous cell carcinoma

Objectives

Locoregional disease recurrence and metastatic events are the leading causes of death and the most important prognostic factors in patients with head and neck squamous cell carcinoma (HNSCC). A major goal of oncology is the identification of clinical and molecular parameters to evaluate the individual risk of recurrence. MicroRNAs (miRNAs) have been shown to correlate well with tumor size and differentiation. Therefore, they are candidate biomarkers for estimating clinical outcomes.

Overexpression and varied clinical significance of Th9 versus Th17 cells in distinct subtypes of oral lichen planus

ObjectiveOral lichen planus (OLP) presents with large numbers of T lymphocytes accumulating beneath the epithelium of the oral mucosa; however, its aetiology remains obscure. A potential role for an emerging novel T cell subset, Th9, in OLP has recently been suggested but remains to be clarified. The current aim was to investigate the expression and potential clinical significance of Th9 cells in distinct subtypes of OLP.Materials and methodsPeripheral blood samples were collected from 41 OLP patients and 18 healthy controls (HCs). Flow cytometric analysis was used to detect the CD4+ T helper subset Th9 (IL-9⁺IL-17⁻CD4⁺ Th cells) and Th17 (IL-9⁻IL-17⁺CD4⁺ Th cells) expression levels.ResultsFlow cytometry results showed significantly elevated levels of Th9 cells in reticular and erosive OLP compared to HCs. Th9 expression in erosive OLP was less than in reticular OLP, indicating that Th9 but not Th17 cells may play a predominant role in reticular disease. However, in erosive OLP patients, we found much higher levels of Th17 cells compared to reticular OLP patients and HCs, indicating that Th17 dominates in erosive OLP. Statistical analysis showed positive correlations of Th9 cells and Th17 cells in patients with reticular or erosive OLP but none in HCs.ConclusionsTh9 and Th17 cells may take the predominant roles in reticular and erosive OLP respectively, and their numbers were positively correlated in reticular and erosive OLP patients. Elevated circulating Th9 cells may help maintain immune balance in OLP immunopathogenesis, which requires further investigation.     

The high expression level of programmed death‐1 ligand 2 in oral lichen planus and the possible costimulatory effect on human T cells

J Oral Pathol Med (2011) 40 : 525–532 Background: Oral lichen planus (OLP) is a T‐cell‐mediated chronic autoimmune disease whose precise etiology is unknown. The recently identified costimulatory programmed death‐1 (PD‐1) molecule and its ligands, PD‐L1 and PD‐L2, have been identified as CD28‐B7 family molecules and constitute a regulatory pathway of potential therapeutic use in immune‐mediated diseases. Methods: We examined the expression of two ligands of PD‐1 at both the protein and gene level in the focal mucosa and peripheral blood of OLP patients using immunohistochemistry and real‐time PCR. Next, we used the PD‐L2.Ig fusion protein and observed its effects on T cells, which were co‐cultured with IFN‐γ‐treated keratinocytes (KCs) in the presence of PHA. Results: We found that the expression of PD‐L2 at both the gene and protein level was statistically different in peripheral blood and local lesion tissue of patients with OLP compared to the normal controls. The proliferation ability of T cells and the expression level of IFN‐γ in the supernatant of the above co‐culture model were significantly augmented (P < 0.05). PD‐L2.Ig fusion protein significantly aggravated the apoptosis of T cells, inhibited the proliferation of T cells and decreased the release levels of IL‐2 and IFN‐γ in the model (P < 0.05). Conclusion: These data show that the increased expression of PD‐L2, as a costimulatory molecule, may have an important modulatory function on the local immune responses of OLP in vivo.