Aberrant DNA methylation of tumor-related genes in oral rinse: a noninvasive method for detection of oral squamous cell carcinoma

BACKGROUND:

The early detection of oral squamous cell carcinoma (OSCC) is important, and a screening test with high sensitivity and specificity is urgently needed. Therefore, in this study, the authors investigated the methylation status of tumor‐related genes with the objective of establishing a noninvasive method for the detection of OSCC.

METHODS:

Oral rinse samples were obtained from 34 patients with OSCC and from 24 healthy individuals (controls). The methylation status of 13 genes was determined by using methylation‐specific polymerase chain reaction analysis and was quantified using a microchip electrophoresis system. Promoter methylation in each participant was screened by receiver operating characteristic analysis, and the utility of each gene's methylation status, alone and in combination with other genes, was evaluated as a tool for oral cancer detection.

RESULTS:

Eight of the 13 genes had significantly higher levels of DNA methylation in samples from patients with OSCC than in controls. The genes E‐cadherin (ECAD), transmembrane protein with epidermal growth factor‐like and 2 follistatin‐like domains 2 (TMEFF2), retinoic acid receptor beta (RARβ), and O‐6 methylguanine DNA methyltransferase (MGMT) had high sensitivity (>75%) and specificity for the detection of oral cancer. OSCC was detected with 100% sensitivity and 87.5% specificity using a combination of ECAD, TMEFF2, RARβ, and MGMT and with 97.1% sensitivity and 91.7% specificity using a combination of ECAD, TMEFF2, and MGMT.

CONCLUSIONS:

The aberrant methylation of a combination of marker genes present in oral rinse samples was used to detect OSCC with >90% sensitivity and specificity. The detection of methylated marker genes from oral rinse samples has great potential for the noninvasive detection of OSCC. Cancer 2012. © 2012 American Cancer Society.     

A minimally invasive immunocytochemical approach to early detection of oral squamous cell carcinoma and dysplasia

Squamous dysplasia of the oral cavity indicates increased risk of progression to squamous cell carcinoma (SCC). An important advance would be the development of a minimally invasive assay for identification of oral SCC and dysplasia. We have investigated the suitability in this context of immunostaining oral smears for minichromosome maintainance proteins (MCMs), sensitive and specific biomarkers of cell cycle entry. Immunohistochemical examination of 66 oral tissue samples showed a greater frequency of Mcm-2 expression in surface layers of moderate/severe dysplasia and SCC compared to benign keratosis/mild dysplasia. Immunocytochemistry for Mcm-2/Mcm-5 was performed on 101 oral smears. Conventional smears included 23 from normal mucosa, benign proliferative disease and mild dysplasia, all of which were MCM negative. Of 52 conventional smears of SCC tissue samples, 18 were inadequate. However, MCM-positive cells were present in 33/34 adequate samples. Of 26 liquid-based cytology smears, 19 out of 20 smears from SCC were adequate and all were MCM positive. Six smears from benign lesions were adequate and MCM negative. We conclude that MCMs are promising markers for early detection of oral SCC and dysplasia, particularly in a liquid-based cytology platform. Detection of MCMs would be amenable to automation and potentially applicable in the developing world. Further studies are now warranted.     

基于纤支镜刷片细胞学的肺癌细胞核形态的多参数定量研究

目的探讨基于纤维支气管镜刷片细胞涂片中,多参数的细胞核形态定量分析对肺癌的鉴别价值。方法采用回顾性的分析方法,利用HMIAS-2000医学图文分析测量系统,对组织学确诊的纤维支气管镜刷片的细胞核进行形态定量研究(腺癌48例、鳞癌28例、小细胞癌22例,阴性对照组40例)。结果在22个形态定量参数中,19项参数有显著统计学差异(P<0.01),2项有统计学差异(P<0.05),1项无统计学差异(P>0.05)。结论多参数的形态计量揭示了纤支镜刷片细胞学的肺癌与肺良性病变细胞核形态学计量特征,对辅助鉴别诊断良恶性具有一定的意义。     

Development and validation of a risk model for noninvasive detection of cancer in oral potentially malignant disorders using DNA image cytometry

Objective:To elucidate whether DNA aneuploidy was an independent discriminator for carcinoma within oral potentially malignant disorders (OPMDs), and further establish and validate a risk model based on DNA aneuploidy for the detection of oral cancer.Methods:A total of 810 consecutive patients with OPMD were prospectively enrolled from March 2013 to December 2018, and divided into a training set (n = 608) and a test set (n = 202). Brushing and biopsy samples from each patient were processed by DNA- DNA image cytometry and histopathological examination, respectively.Results:DNA aneuploidy of an outside DNA index ≥ 3.5 in OPMD was an independent marker strongly associated with malignant risk [adjusted odds ratio: 13.04; 95% confidence interval (CI): 5.46–31.14]. In the training and test sets, the area under the curve (AUC) was 0.87 (95% CI: 0.82–0.91) and 0.77 (95% CI: 0.57–0.97), respectively, for detecting carcinoma in OPMD patients. The independent risk factors of lateral/ventral tongue and non-homogenous type combined with a risk model built with a multivariate logistic regression revealed a more favorable diagnostic efficacy associated with the training set (AUC: 0.93; 95% CI: 0.91–0.96) and test set (AUC: 0.94; 95% CI: 0.90–0.98). The sensitivity and specificity of carcinoma detection within OPMD was improved to 100% and 88.1%, respectively.Conclusions:This large-scale diagnostic study established a risk model based on DNA aneuploidy that consisted of a noninvasive strategy with lateral/ventral tongue and non-homogenous features. The results showed favorable diagnostic efficacy for detecting carcinoma within OPMD, irrespective of the clinical and pathological diagnoses of OPMD. Multicenter validation and longitudinal studies are warranted to evaluate community practices and clinical applications.     

Monitoring of circulating tumour-associated DNA as a prognostic tool for oral squamous cell carcinoma

Frequent allelic imbalances (AIs) including loss of heterozygosity and microsatellite instability on a specific chromosomal region have been identified in a variety of human malignancies. The objective of our study was to assess the possibility of prognostication and monitoring of oral squamous cell carcinoma (SCC) by microsatellite blood assay. DNA from normal and tumorous tissues and serum DNA obtained at three time points (preoperatively, postoperatively, and 4 weeks postoperatively) from 64 patients with oral SCC was examined at nine microsatellite loci. In all, 38 (59%) DNA samples from tumorous tissues and 52% from serum showed AIs in at least one locus. Patterns of AIs in the serum DNA were matched to those detected in tumour DNA. Of them, AIs were frequently detected preoperatively (44%, 28 of 64), and postoperatively (20%, 13 of 64). Moreover, among 12 cases with AIs during the postoperative period, six had no evidence of an AI 4 weeks postoperatively, and they had no recurrence and were disease free. In contrast, six patients with AI-positive DNA 4 weeks postoperatively have died with distant metastasis within 44 weeks. Thus, our results suggest that the assessment of microsatellite status in the serum DNA could be a useful predictive tool to monitor disease prognosis.     

Sentinel lymph node biopsy in squamous cell carcinomas of the lips and the oral cavity - a single center experience

BACKGROUND AND OBJECTIVES: Despite the availability of initial data on sentinel lymph node biopsy (SLNB) in OSCC, its value in clinical practice remains unclear. The aim of this study is to evaluate the feasibility and potential of SLNB as a reliable staging procedure in early stage SCC of the lips and the oral mucosa. METHODS: From 2001 to 2004, 40 patients with early stage SCC of the lips and oral cavity were enrolled. The main inclusion criteria were: cN0 category by MRI and ultrasound, tumor category T1-T2, for certain sites also T3. Patients with positive SLN underwent MRND and/or radiation. Patients with negative SLN were monitored monthly. RESULTS: In eight patients, the SLN were positive. Two patients with negative SLN subsequently developed nodal disease (5% false negatives). Twenty-one patients have a follow-up longer than 24 months. Five patients died (one from nodal disease, one from local recurrence, and three from unrelated causes). One patient developed a second primary tumor, the remaining patients are free of disease. CONCLUSIONS: SLNB may contribute to more targeted lymph node dissection strategies. Further studies will have to ensure the safety of this procedure in comparison to selective neck dissection.     

口腔鳞癌细胞中ITGB6基因主要转录调控区域的定位分析

目的： 鉴定ITGB6基因在口腔鳞癌细胞中的主要调控区域,为研究ITGB6基因在口腔鳞癌细胞中的转录调控机制奠定基础。方法：构建含有ITGB6基因转录起始点上游5' 端侧翼区不同长度DNA序列的重组pGL2荧光素酶报告基因质粒,转染口腔鳞癌细胞株TCA8113和SAS,利用双荧光素酶报告基因系统检测启动子转录活性,并通过生物信息学方法预测ITGB6基因中潜在的主要转录因子结合位点。结果：获得一系列不同长度ITGB6基因5' 侧翼区序列的重组荧光素酶报告基因质粒;当ITGB6基因5' 端侧翼片段截短到-187～-35和-35～+27之间时,启动子活性均显著下降;生物信息学分析显示,在-187～-35区域有2个Ets-1的潜在结合位点,而在-35～+27区域有1个IRF-4潜在结合位点。结论：在口腔鳞癌细胞中,ITGB6基因-187~-35和-35~+27区域是主要的转录因子结合区。     

早期口腔鳞癌颈部淋巴结转移的研究进展

口腔鳞癌具有较高的隐匿性颈淋巴结转移率,然而,临床上对早期患者颈部淋巴结的处理还存在较大争议.从肿瘤原发灶浸润厚度、影像学方面探讨早期口腔鳞癌颈部淋巴结转移的因素.对肿瘤原发灶浸润厚度>4 mm的患者,结合肿瘤影像学检查,建议行选择性颈部淋巴结清扫术.     

皮肤镜图像分析技术用于白癜风的诊断与疗效观察

目的探讨皮肤镜图像分析技术在白癜风的早期诊断与疗效观察中的研究。方法收集来院就诊患者,分别经肉眼初诊及皮肤镜偏振法（DP）和皮肤镜浸润法（DS）诊断和疗效评价,对诊断不清者可酌情行组织病理确诊; 并比较DP和DS两种方法的性能。结果在93例白癜风皮损中,进展期59例,其毛周色素残留发生率达91.67%,显著高于稳定期34例的58.33%,两者之间差异具有统计学意义（P〈0.05）; 在进展期毛囊周围色素残留和经治疗后早期色素岛形成的皮损复色迹象发生率达77.28%,明显高于稳定期45.46%。结论在白癜风皮损中发现毛囊周围色素残留是其特异性表现,并可观察到白癜风康复期的早期复色迹象,该研究对白癜风的早期诊疗与疗效判断、减轻患者精神压力、增加治疗信心及提高总体疗效有重要意义。     

Diagnostic value of lncRNAs as potential biomarkers for oral squamous cell carcinoma diagnosis: a meta-analysis

Objective Several studies have revealed the critical role of long non-coding RNAs (lncRNAs) as biomarkers for diagnosing oral squamous cell carcinoma (OSCC). However, the data remain inconsistent. This meta-analysis was performed to summarize the potential of lncRNAs as OSCC biomarkers. Methods We searched PubMed, Cochrane Library, Web of Science, and China National Knowledge Infrastructure databases for literature published until December 10, 2020. Study quality was assessed using Quality Assessment for Studies of Diagnostic Accuracy-2, and sensitivity, specificity, and other measures regarding lncRNAs for OSCC diagnosis were pooled using bivariate meta-analysis models. Data analyses were performed using STATA 14.0. Results Overall, 8 studies with 981 cases and 585 controls were included in the pooled analysis. The pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio, and area under the receiver operating characteristic curve values were as follows: 0.76 [95％ confidence interval (CI), 0.65–0.84], 0.90 (95％ CI, 0.82–0.95), 7.5 (95％ CI, 4.20–13.40), 0.27 (95％ CI, 0.18–0.39), 28 (95％ CI, 13.00–58.00), 0.90 (95％ CI, 0.87–0.93), respectively. Deeks' funnel plot asymmetry test (P = 0.56) indicated no potential publication bias. Conclusion Our meta-analytical evidence suggests that lncRNAs could be employed as a potential non-invasive diagnostic tool for OSCC.     

早期舌鳞癌复发原因及预后因素的分析

背景与目的：早期舌癌的治疗效果理想,但复发是影响早期舌癌治疗效果的重要因素。本研究回顾性分析227例早期舌癌的临床资料,探讨早期舌癌复发的主要原因及影响预后的因素。方法：收集中山大学肿瘤防治中心1992—2003年首治的227例早期舌鳞癌患者的临床资料,Kaplan—Meier法对各影响因素进行生存率分析并计算生存率,log-rank法比较各因素不同水平生存分布的差异。Cox同归模型进行多因素分析,影响复发不同因素间比较用卡方检验。结果：高分化鳞癌组复发率为19.3％,中高低分化鳞癌组复发率为39．2％,二者的差异具有统计学意义（P=0．004）,综合治疗组复发率为15％,单纯手术组复发率为28％．二者的差异具有统计学意义（P=0．014）。复发组及非复发组3年及5年生存率分别为40．7％、87.3％及25．9％、80．3％,二者的差异具有统计学意义（P=0.000）。行颈清扫组及未行颈清扫组3年及5年生存率分别为为76．4％、75．0％及68％、61．2％．二者的差异具有统计学意义（P=0．023）;≥45岁组及〈45岁组3年及5年生存率分别为75．6％、78％及59．5％、66．1％,二者的差异具有统计学意义（p=0．021）。结论：病理分级及是否进行综合治疗是影响早期舌癌复发的原因,患者的年龄、是否行颈清扫、是否复发是影响早期舌癌预后的因素,年龄及是否复发是影响预后的独立因素。