HLA typing in Taiwanese patients with oral submucous fibrosis

BACKGROUND: A significant association of certain human leukocyte antigens (HLA) and haplotypic pairs with oral submucous fibrosis (OSF) has been reported. However, controversial result of no HLA association with OSF has also been reported. In this study, the phenotype and haplotype frequencies of HLA-A, -B, -C, -DRB1, and -DQB1 in 135 Taiwanese OSF patients were calculated and compared with those in 540 healthy control Taiwanese. METHODS: The analysis of HLA-A, -B, and -C antigens, and of HLA-DRB1 and -DQB1 alleles in OSF patients and healthy control subjects, was performed by serologic typing and DNA typing using polymerase chain reaction with sequence-specific primers (PCR-SSP), respectively. RESULTS: We found that the phenotype frequency of HLA-B76 (3.0%) in OSF patients was significantly greater than that (0%) in healthy control subjects (corrected P (Pc) = 0.000). In addition, the haplotype frequencies of HLA-B48/Cw7 (3.0%), -B51/Cw7 (6.7%), and -B62/Cw7 (8.2%) in OSF patients were significantly greater than the corresponding haplotype frequencies (0, 0.7, and 1.9%, respectively) in healthy control subjects (Pc = 0.000). The relative risk (RR) values of haplotypes B51/Cw7 (9.57) and B62/Cw7 (4.7) were greater than the RR values of phenotypes B51 (1.81), B62 (2.31), and Cw7 (1.91) in OSF patients. In addition, the etiologic fraction (EF) value of haplotype B51/Cw7 (0.63) was higher than the EF values of phenotypes B51 (0.2) and Cw7 (0.59). CONCLUSIONS: We conclude that some Taiwanese areca quid (AQ) chewers with particular HLA phenotypes and haplotypes are prone to have OSF. In addition, some particular HLA haplotypes may play more important roles than the individual HLA phenotypes for the genetic susceptibility to OSF. However, the significantly increased HLA phenotype B76 and three of the common HLA haplotypes detected are present in only about 20% of incident cases of OSF.     

High incidence of autoantibodies in Taiwanese patients with oral submucous fibrosis.

BACKGROUND: Previous study has shown a high incidence of autoantibodies including antinuclear (ANA), antismooth muscle (SMA), antigastric parietal cell (GPCA), antithyroid microsomal (TMA), and antireticulin antibodies in a small group of 26 patients with oral submucous fibrosis (OSF). The reasons why some of the OSF patients have high titers of autoantibodies in serum have not been completely explained and no further study on autoantibodies in OSF patients has been done in a large group of patients. METHODS: In this study, we determined the serum levels of ANA, SMA, GPCA, and TMA in a large group of 109 male Taiwanese patients with OSF by an indirect immunofluorescence technique (for ANA, SMA, and GPCA), and by a semiquantitative microtiter particle agglutination test (for TMA). The presence of serum autoantibodies in OSF patients was further correlated with patients' oral habits and the severity of OSF measured by maximum mouth opening (MMO) and sites of involvement. RESULTS: We found that the frequencies of presence of serum ANA (23.9%), SMA (23.9%), and GPCA (14.7%) in OSF patients were significantly higher than those (9.2, 7.3, and 5.5%, respectively) in healthy control subjects (P < 0.01, P < 0.005, and P < 0.05, respectively). Although the frequency of presence of TMA (5.5%) in OSF patients was also greater than that (2.8%) in healthy control subjects, the difference was not significant (P > 0.05). The presence of serum GPCA in OSF patients was significantly associated with daily areca quid (AQ) consumption (P < 0.05). The presence of serum ANA in OSF patients associated with daily AQ consumption was of borderline statistical significance (P = 0.066). However, no significant correlations were demonstrated between the presence of serum autoantibodies in OSF patients and other variables of oral habits, MMO, and sites of involvement. CONCLUSION: In this study, all the 109 OSF patients had AQ chewing habit and 73.4% of the OSF patients swallowed the 'juice' of AQ during the chewing process. The presence of serum GPCA and ANA in OSF patients was associated with daily consumption of AQs. AQ chewing caused mucosal microtrauma, and ulcerations facilitated the diffusion of genotoxic and cytotoxic AQ ingredients into the oral and gastric tissues. Altered autoantigens released from AQ ingredients-damaged cells may induce autoantibody production. Higher frequencies of specific HLA-DR antigens in OSF patients may also help autoantibody production. Therefore, we conclude that the high incidence of autoantibodies in OSF patients may be due to AQ chewing habit, toxic AQ ingredients, and genetic susceptibility of the OSF patients.     

Growth of oral and skin fibroblasts from patients with oral submucous fibrosis

The purpose of the investigation was to evaluate and compare the proliferation (growth) of mouth fibroblasts and skin fibroblasts from patients with oral submucous fibrosis (OSF). Material comprised fibroblasts from fibrous bands situated in the buccal mucosa and from the inner aspect of the forearm of 8 patients with classic features of OSF as well as fibroblasts from 6 buccal mucosa and 8 skin biopsy specimens from healthy non-areca nut chewing individuals. Cells were cultured for 8 days according to standard techniques. Their growth was monitored daily, under optimal conditions as well as exposure to concentrations of arecoline. The data were analyzed using regression analysis, analysis of variance and the Kruskal-Wallis test. We found no statistically significant differences between the proliferation patterns of oral and skin fibroblasts from patients or between those from patients and controls. The reaction of the cells exposed to concentrations of arecoline was similar; at low concentrations (0.1–10 μg/ml) normal growth was maintained, while 100 μg/ml inhibited growth. It is concluded that fibroblasts from mouths affected by OSF have proliferation patterns which fall within normal parameters, that the excessive collagen formation in established OSF is not due to increased fibroblast proliferation and that arecoline does not stimulate fibroblast proliferation.     

Relationship between thickness of fibrosis and epithelial dysplasia in oral submucous fibrosis

Abstract Aim: Although oral submucous fibrosis is characterized by fibrosis of the subepithelial connective tissue, the overlying epithelial changes contribute to malignant transformation. Therefore, the aim of the study was to evaluate the relationship between thickness of fibrosis and epithelial changes in oral submucous fibrosis. Methods: The relationship between thickness of fibrosis and presence or absence of epithelial dysplasia was evaluated in 107 biopsies containing histopathologically‐confirmed oral submucous fibrosis. The results were analyzed using Student’s t‐test or χ²‐test. Results: Fifty‐seven percent (61/107) of oral submucous fibrosis lesions showed a non‐dysplastic overlying epithelium, while 43% (46/107) showed varying degrees of epithelial dysplasia. The mean thickness of fibrosis of non‐dysplastic lesions was 0.91 ± 0.41 mm (mean ± standard deviation) and ranged from 0.25 to 1.9 mm. However, the mean thickness of fibrosis of dysplastic lesions was 1.17 ± 0.52 mm and ranged from 0.48 to 3 mm. The results revealed a significant increase in the incidence of epithelial dysplasia as the thickness of fibrosis increased (P = 0.004). As such, the lesions that showed increased fibrosis were more likely to present with epithelial dysplasia. Conclusions: The advancement of fibrosis increases the risk of development of epithelial dysplasia in oral submucous fibrosis.     

口腔黏膜下纤维性变并存白斑74例的临床病理分析

目的：???观察口腔黏膜下纤维性变（OSF）并存白斑（OLK）患者的临床表现以及病理特征,探讨其上皮异常增生程度与纤维性变严重程度的相关性。方法整理74例OSF并存OLK患者的临床病理资料并进行分析。结果74例OSF并存OLK患者多见于中青年男性,均有咀嚼槟榔史,以20~49岁年龄段发病率较高,临床上同时具有OSF、OLK的临床症状和体征。病理表现为上皮细胞间隙增宽;基底膜增厚、不规则;胶原纤维水肿,变性。结论 OSF并存OLK患者的上皮异常增生程度与纤维性变严重程度无显著相关性;OSF并存OLK并非两种疾病的单纯叠加,而具有其特征性的表现。     

Fingerprinting genomic instability in oral submucous fibrosis

Background:  Oral submucous fibrosis (OSF) is a high-risk pre-cancerous condition where 7–13% of these patients develop head and neck squamous cell carcinoma (HNSCC). To date there is no cancer predictive markers for OSF patients. Genomic instability hallmarks early genetic events during malignant transformation causing loss of heterozygosity (LOH) and chromosomal copy number abnormality. However, to date there is no study on genomic instability in OSF. Although this condition is known as a high-risk pre-cancerous condition, there is no data regarding the genomic status of this disease in terms of genetic susceptibility to malignant transformation.
Methods:  In this study, we investigated the existence of genetic signatures for carcinogenesis in OSF. We employed the high-resolution genome-wide Affymetrix Mapping single nucleotide polymorphism microarray technique to 'fingerprint' global genomic instability in the form of LOH in 15 patient-matched OSF–blood genomic DNA samples.
Results:  This rapid high-resolution mapping technique has revealed for the first time that a small number of discrete hot-spot LOH loci appeared in 47–53% of the OSF tissues studied. Many of these LOH loci were previously identified regions of genomic instability associated with carcinogenesis of the HNSCC.
Conclusion:  To our knowledge, this is the first evidence that genomic instability in the form of LOH is present in OSF. We hypothesize that the genomic instability detected in OSF may play an important role in malignant transformation. Further functional association studies on these putative genes may reveal potential predictive oral cancer markers for OSF patients.     

OSF并存OLP34例回顾性分析

目的：总结口腔黏膜下纤维性变（OSF）并存口腔扁平苔藓（OLP）的临床特征,提高对该病的诊断和治疗水平。方法：对34例OSF并存OLP患者的临床资料进行回顾性分析。结果：34例OSF并存OLP患者表现为青壮年男性居多,均有进食刺激性食物和咀嚼槟榔史,29例（85.29%）有吸烟史,27例（79.41%）有饮酒史,28例（82.35%）OSF的病理分期为早期,6例（17.65%）为中期,发病部位以颊部和舌部为主,所有患者均无糜烂和张口受限,治疗后疼痛症状缓解,但白色条纹或斑点始终未消退,已消失的丝状乳头、菌状乳头亦未见恢复。结论：OSF并存OLP并非两种疾病的简单叠加,而是具有其特异性的表现,必要时可行两个典型部位活检进行诊断,治疗中及治疗后须戒烟、戒槟榔。     

Predictors of the severity of oral submucous fibrosis among gutka consumers: a regression analysis

Oral submucous fibrosis (OSMF) is an insidious chronic disease of the oral mucosa that is characterised by severely limited mouth opening, blanching of the oral mucosa, and a burning sensation in the oral cavity. Consumption of betel nut and/or gutka are the known risk factors. We undertook this study to correlate the frequency and duration of gutka intake with the severity of OSMF and to determine the predictors of severe OSMF (mouth opening <20mm). A cross sectional study was conducted on 300 participants (who were known gutka chewers) selected at the Baqai Dental College and Fatima Hospital, Karachi, Pakistan. Participants’ medical and dental histories were recorded. Informed consent was obtained, and clinical oral examination was done. Information regarding the extent of mouth opening, chewing habits, frequency and duration of gutka intake, the site of placing gutka, duration of chewing, and whether they swallowed or spat out the gutka were collected. A standardised questionnaire was used to document the findings. Binary logistic regression was applied using the severity of OSMF (mouth opening <20mm) as an outcome variable. Out of 300 participants, 172 (57.3%) were males; mean (SD) age of the sample was 38.2 (12.3) years. A total of 156 (52%) participants had the habit of chewing gutka, of which 213 (71%) had clinical stage I OSMF, 75 (25%) had stage II, and 12 (4%) had stage III. As per functional staging, 18 (6%) subjects had mouth opening <20mm. Nearly 144 (48%) participants were consuming other forms of tobacco in addition to the gutka. Patients with palpable bands = 232 (77.3%), ulcerative lesions = 212 (70.7%), altered taste sensation = 210 (70%) and altered hunger = 252 (85.7%) were common findings. The duration of gutka intake was found to be positively correlated with the severity of OSMF; however; its frequency was not. Among intraoral findings, the presence of red and white lesions inside the mouth was the most significant predictor of the severity of OSMF.     

口腔黏膜下纤维性变组织中Smad7蛋白的表达研究

目的检测Smad7蛋白在口腔黏膜下纤维性变(OSF)组织中的表达及分布,探讨其在OSF发病机制中的作用。方法采用免疫组化抗生蛋白链菌素生物素复合体法(strept Avidin-Biotin complex,SABC),用Smad7兔抗人多克隆抗体检测20例OSF病变组织及10例正常口腔黏膜组织中的Smad7蛋白的表达及分布。结果Smad7在OSF病变组织中为弱阳性表达,在正常口腔黏膜组织中为阴性表达,差别具有显著意义(P<0.05)。结论Smad7蛋白在OSF病变组织中弱阳性表达,在OSF发病机制中有重要作用。     

口腔黏膜下纤维化中VEGF mRNA的表达研究

目的:研究口腔黏膜下纤维化(oral submucous fibrosis,OSF)患者早、中、晚期组中血管内皮生长因子(vascular endothelial growth factor,VEGF)mRNA的表达,探讨VEGF与OSF微血管病变发生发展的关系.方法:选取OSF患者30例,其中早、中、晚期组各10例,5例健康志愿者为对照组.每例研究对象取其1:3腔黏膜,运用逆转录聚合酶链反应(RT-PCR)方法,研究OSF患者早、中、晚期组中VEGF mRNA的表达水平.结果:VEGF mRNA在OSF早、中期的表达均较正常口腔黏膜高(P＜0.05):VEGF mRNA在OSF早期的表达均明显高于中、晚期(P＜0.05),其中中期的表达与晚期相比差异无显著性(P＞0.05).结论:VEGF对OSF微血管病变的影响主要在黏膜下层,各种调控因素在基因转录水平导致VEGF的高表达可能是OSF局部组织缺血的代偿.     

口腔黏膜下纤维性变中MMP-2基因的表达及意义

目的：检测口腔黏膜下纤维性变中基质金属蛋白酶-2（MMP-2）的表达并探讨其病理意义.方法：抽提11例口腔黏膜下纤维化组织和10例正常口腔黏膜组织的总RNA,通过逆转录聚合酶链反应（PF-PCR）检测MMP-2mRNA在口腔黏膜下纤维性变患者颊黏膜中的表达并与正常口腔黏膜进行比较.结果：口腔黏膜下纤维性变患者颊黏膜组织中MMP-2 mRNA表达高于正常颊黏膜（p＜0.05）.结论：MMP-2基因的表达与口腔黏膜下纤维性变组织重塑过程密切相关.     

TLR2在口腔黏膜下纤维性变组织中的表达及意义

目的：通过检测口腔黏膜下纤维性变（OSF）组织中TLR2的表达情况,初步探讨TLR2在OSF发病中的作用。方法：选取OSF患者30例（早、中、晚期各10例）为实验组,正常者5例为对照组,每例研究对象取其口腔颊黏膜,采用免疫组化SABC法检测各组织中TLR2的表达情况。结果：正常口腔颊黏膜组织中TLR2呈阳性表达,在OSF组织中TLR2表达明显增强（P〈0.05）,主要表达于黏膜下组织;OSF早、中、晚期组织中TLR2的表达无显著性差异（P〉0.05）。结论：与正常口腔黏膜相比,OSF组织中TLR2的表达增高,提示TLR2在OSF发病中起一定的促进作用。     

Comprehensive analysis of microRNAs and their target genes in oral submucous fibrosis

The objective of the study is to understand the role of experimentally validated microRNAs (miRNAs) contributing to the acquisition of oncogenic phenotype in oral submucous fibrosis (OSF) by computational analysis. A comprehensive review was carried out to corroborate and summarize altered miRNA expression in OSF by retrieving relevant publications querying MEDLINE, Web of Science, Embase, and Scopus. The association between the miRNA-mRNA was performed using miRTarBase 8.0. The visualization of the miRNA-mRNA interaction was plotted using Cytoscape. MIENTURNET was used for the pathway analysis. Enrichment analysis was carried out for elucidating the hierarchical functions of miRNAs related to the acquisition of biological processes involved in the development of cancer. Thirteen miRNAs (hsa-miR-499a, hsa-miR-200b, hsa-miR-200c, hsa-miR-1246, hsa-miR-31, hsa-miR-10b, hsa-miR-21, hsa-miR-203, hsa-miR-455, hsa-miR-760, hsa-miR-623, hsa-miR-610, and hsa-miR-509-3-5p) were found to be deregulated in OSF. A total of 371 experimentally validated genes were shown to be interacting with the OSF-associated miRNAs. The targets of antifibrotic and profibrotic miRNAs were enriched in the cancer-related pathways. Dysregulated miRNA and its target genes illustrate the physiological role of miRNAs in fibrosis. Understanding the miRNA-mediated fibrotic signaling and targetting the specific miRNA-target gene interaction might provide relevant cues to ameliorate the fibrotic disease.     

口腔黏膜下纤维性变组织中Smad2/3蛋白的表达研究

目的：检测Smad2／3蛋白在13腔黏膜下纤维性变（OSF）组织中的表达及分布,探讨其在OSF发病机制中的作用。方法：采用免疫组化SABC法,用Smad2／3兔抗人多克隆抗体检测20例OSF病变组织及10例正常13腔黏膜组织中的Smad2／3蛋白的表达及分布。结果：Smad2／3在OSF病变组织中为中等强度表达,在正常口腔黏膜组织中为强阳性表达,具有显著性差异（P〈0．05）。结论：Smad2／3蛋白在OSF病变组织中中等强度表达,在OSF发病机制中有重要作用。