Silencing KLF16 inhibits oral squamous cell carcinoma cell proliferation by arresting the cell cycle and inducing apoptosis

Krüppel-like factor 16 (KLF16), a member of the Krüppel-like factor (KLF) family, has been extensively investigated in multiple cancer types. However, the role of KLF16 in oral squamous cell carcinoma (OSCC) remains unknown. Thus, we conducted this study to investigate its related mechanism. KLF16 expression in OSCC cell lines was quantified by western blotting. Then, OECM1 and OC3 cells were divided into Blank, siCtrl, siKLF16#1 and siKLF16#2 groups. Subsequently, cell proliferation was detected using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assays, cell migration and invasion were detected with wound healing and Transwell assays, and cell cycle distribution and cell apoptosis were detected via flow cytometry. KLF16, p21, CDK4, Cyclin D1 and p-Rb expression was detected by western blotting. Finally, xenograft models were established in nude mice to observe the in vivo effects of KLF16 on OSCC. KLF16 protein expression was upregulated in OSCC cells. Compared to the cells in the Blank group, the OECM1 and OC3 cells in the siKLF16#1 group and siKLF16#2 group exhibited a sharp decrease in proliferation but a remarkable increase in apoptosis. Moreover, the proportion of cells in the G0/G1 phase notably increased and that in the S phase decreased, with evident decreases in cell invasion and migration. Moreover, KLF16, cyclin-dependent kinase 4 (CDK4), Cyclin D1 and p-Rb protein expression was upregulated, but p21 expression was downregulated. The mice in the siKLF16#1 and siKLF16#2 xenograft model groups exhibited slower tumour growth and smaller tumours with evident downregulation of Ki67 expression compared to the mice in the Blank group. KLF16 expression was upregulated in OSCC cells, and interfering with KLF16 led to cell cycle arrest, inhibited OSCC cell growth and promoted cell apoptosis.     

Preoperative assessment of the risk of postoperative death in patients with oral squamous cell carcinoma: a consideration beyond age,sex, and stage of cancer

Despite improvements in the management of patients in critical care, about 3% patients who have an operation with curative intent for oral squamous cell carcinoma (SCC) do not survive their stay in hospital. Our aim was to assess the risk factors for postoperative death that were independent of the stage of the cancer, or the age and sex of the patients. We screened 4760 consecutive inpatients at a maxillofacial tertiary care centre from 2011 to 2016, and 34 of them had died within the first three months after operation. We matched them with a further 34 patients with the same TNM stage, age, and sex. General personal and clinical data and preoperative laboratory values were screened, and we applied a Charlson Comorbidity Score (for anaesthetic risk) for each group. Patients’ mean (SD) age was 66 (12) years old. There was no significant difference in sex (p = 1), age (p = 0.718), or TNM classification. Those who died after operation had significantly more renal (p = 0.027) and gastrointestinal (p = 0.006) diseases, but cardiac diseases (p = 0.468) and diabetes mellitus (p = 1) were not significant risk factors in themselves. Patients who died postoperatively had significantly worse risk scores (p = 0.001) overall. The most common causes of death were septic shock (n = 10) and acute cardiac (n = 9) or respiratory failure (n = 7). Our findings suggested that general diseases were not intrinsically a contraindication for operation with curative intent. The Charlson Comorbidity Score helped to detect potentially fatal courses and could be useful in the preoperative assessment of patients whose general health is not good.     

PD1 expression and correlation with its ligands in oral cancer specimens and peripheral blood

ObjectivesThis study analyzed the expression of the PD1 receptor in tumor tissue and peripheral blood of oral squamous cell carcinoma (OSCC) patients, and correlated it with the PD1 ligands PD-L1 and PD-L2. The currently low response rates of checkpoint inhibitor treatment in OSCC could be increased by a better understanding of immune checkpoint biology. Despite evidence in the literature for upregulation of PD1 checkpoint ligands in OSCC tissue, there has been no correlation analysis of the PD1 receptor with its ligands in tissue specimens and peripheral blood of OSCC patients.Materials and methodsAn RT-qPCR analysis of PD1 mRNA expression was performed in oral cancer specimens, healthy mucosa, and corresponding blood samples. A cut-off point (COP) was determined and a chi-square (χ²) test was carried out. PD1 expression was correlated with previously reported PD-L1 and PD-L2 expression values using the Spearman test.ResultsTissue and blood specimens of 48 OSCC patients and 26 healthy individuals were analyzed. PD1 expression in OSCC specimens was significantly increased (p = 0.006) compared with healthy oral mucosa. PD1 overexpression in tissue samples showed a significant association with the presence of malignancy (p = 0.006). PD1 expression in tissue samples showed a significant positive correlation (p < 0.001) with the ligands PD-L1 and PD-L2. In contrast, there was no correlation between PD1 and its ligands in blood samples. However, there was a significant positive correlation (p < 0.001) between the ligands PD-L1 and PD-L2, both in tissue and blood samples.ConclusionsIncreased PD1 expression might be a manifestation of T-cell exhaustion in OSCC specimens, leading to immune tolerance. PD-L1/PD-L2-PD1 interaction may be a major mediator of local immunosuppression in OSCC, requiring advanced multimodal treatment protocols.     

Management of the clinically N0 neck in early-stage oral squamous cell carcinoma (OSCC). An EACMFS position paper

Metastasis of oral squamous cell carcinoma (OSCC) to the cervical lymph nodes has a significant impact on prognosis. Accurate staging of the neck is important in order to deliver appropriate treatment for locoregional control of the disease and for prognosis.The management of the neck in early, low volume disease (clinically T₁/T₂ oral cavity tumours) has long been debated. The risk of occult nodal involvement in cT₁/T₂ OSCC is estimated around 20–30%.We describe the natural evolutionary history of OSCC and its patterns of spread and metastasis to the local lymphatic basins. We discuss most published literature and studies on management of the clinically negative neck (cN₀). Particular focus is given to prospective randomized trials comparing the outcomes of upfront elective neck dissection against the observational stance, and we summarize the results of the sentinel node biopsy studies.The paper discusses the significance of the primary tumour histological characteristics and specifically the tumour's depth of invasion (DOI) and its impact on predicting nodal metastasis. The DOI has been incorporated in the TNM staging highlighting its significance in aiding the treatment decision making and this is reflected in world-wide oncological guidelines.The critical analysis of all available literature amalgamates the existing evidence in early OSCC and provides recommendations in the management of the clinically N₀ neck.     

甲苯胺蓝染色在口腔潜在恶性疾患及口腔鳞状细胞癌早期诊断中的临床应用价值研究

目的　综合评价甲苯胺蓝染色在口腔潜在恶性疾患（oral potential malignant disorders,OPMDs）及口腔鳞状细胞癌（oral squamous cell carcinoma,OSCC）早期诊断中的临床应用价值。方法　选取2016年8月至2018年5月于北京大学口腔医院口腔黏膜科就诊的OPMDs及OSCC患者533例。对所有患者进行临床检查、甲苯胺蓝染色和组织病理学检查。以组织病理学诊断为金标准,评价甲苯胺蓝染色的灵敏度、特异度和准确度;绘制受试者工作特征曲线（receiver operator characteristic curve,ROC）及计算曲线下面积（area under curve,AUC）,评价甲苯胺蓝染色的诊断价值;并对上皮异常增生或OSCC的影响因素进行logistic回归分析。结果　甲苯胺蓝染色诊断上皮异常增生或OSCC的灵敏度为75.32%,特异度为84.17%,准确度为81.61%;诊断OSCC的灵敏度为97.10%,特异度为76.51%,准确度为79.17%;诊断上皮异常增生的灵敏度为57.65%,特异度为84.17%,准确度为79.31%。随着上皮异常增生程度的加重,甲苯胺蓝染色诊断的灵敏度越高;且甲苯胺蓝染色诊断轻、中度上皮异常增生的灵敏度均高于临床危险度诊断,其差异均有统计学意义（均P < 0.05）。ROC曲线分析,甲苯胺蓝染色诊断上皮异常增生或OSCC的AUC值为0.797,高于临床危险度诊断的AUC值（0.721）;甲苯胺蓝染色诊断OSCC的AUC值为0.868,高于临床危险度诊断的AUC值（0.802）。logistics回归分析发现,年龄、病程、活检病损部位、临床危险度和甲苯胺蓝染色均与上皮异常增生或OSCC显著相关,甲苯胺蓝染色阳性者患有上皮异常增生或OSCC的风险是甲苯胺蓝染色阴性的13.655倍。结论　综合多项指标评估结果,甲苯胺蓝染色是一种诊断效力强的无创辅助检查技术,推荐应用于OPMDs及OSCC的早期诊断。     

β-catenin及c-Myc蛋白表达与口腔鳞状细胞癌相关性研究

目的 探讨β-catenin和c-Myc在原发性口腔鳞癌(OSCC)中的表达及意义.方法 用免疫组化法检测口腔鳞癌及癌旁正常组织中3-catenin和c-Myc的表达.结果 β-catenin在口腔鳞癌、相应癌旁正常组织中异常表达率分别为56.92％、15.38％,两组之间差异具有统计学意义(P =0.006),β-catenin异常表达增多与肿瘤大小(P=0.003)、局部的淋巴结转移密切相关(P=0.017).在口腔鳞癌、相应癌旁正常组织中c-Myc的阳性表达率为64.62％,7.69％,两组之间差异具有统计学意义(P=0).β-catenin在胞质的表达与c-Myc的表达呈正相关(rs =0.237,P=0.049).结论 c-Myc在口腔鳞癌中过表达,作为原癌基因促进肿瘤发生;β-catenin胞浆异常表达可能与口腔鳞癌的侵袭转移有关.     

MAC30蛋白在口腔鳞状细胞癌中的表达及其临床意义

检测20例正常口腔黏膜和43例口腔鳞状细胞癌患者MAC30蛋白的表达情况。MAC30蛋白在口腔鳞状细胞癌中的表达较正常黏膜明显增加(P<0.05);低分化组和淋巴结转移组较高分化组及淋巴结无转移组阳性表达率明显增高(P<0.05);MAC30蛋白在口腔鳞状细胞癌的发生、发展、分化及转移过程中可能起着重要作用,并可作为一个预测肿瘤转移的新的分子生物学指标。     

OSCC中ER-α、RAR-β、MGMT。及P16INK4a基因启动子甲基化的临床研究

目的：探讨肿瘤相关基因ER-α、RAR-β、MGMT及P16INK4a启动子在口腔鳞状细胞癌（OSCC）组织中的甲基化状态。方法：20例病理确诊为OSCC的组织切片,经酶消化法提取组织DNA后双硫法检测ER-αL、RAR-β、MGMT及P16INK4a基因启动子的甲基化状态,比较分析4种基因启动子甲基化状态和临床病理参数的相关性。结果：20例中,P16INK4a、MGMT启动子甲基化发生率均为10％,RAR-B启动子甲基化发生率为40％,ER-d启动子甲基化发生率为55％,两株OSCC细胞系中,ER-α、RAR-β启动子均出现甲基化,而MGMT及P16INK4a启动子均未见甲基化。结论：RAR-β、ER-α基因启动子的甲基化较P16INK4a、MGMT＇g更为常见,提示前两者可能在OSCC的发生中具有更重要的作用。