Artepillin C as a targeting survivin molecule in oral squamous cell carcinoma cells in vitro: A preliminary study

Background

Survivin, a member of the inhibitor of apoptosis family, is overexpressed in most human tumors, but undetectable in normal adult tissues. It is a promising target molecule in cancer treatment, as interference in its function promotes apoptosis. Artepillin C, a major, biologically active ingredient of Brazilian propolis, possesses anticancer activity against several cancer cells with different tissue origins. However, little is known about its bioactivity on oral squamous cell carcinoma cells or its effect on survivin expression. The aim of this study was to investigate the cytotoxic and antisurvivin activities of artepillin C in oral squamous cell carcinoma cells.

Methods

HSC‐3 human oral squamous cell carcinoma cells were treated with varying doses of artepillin C for up to 72 hours. Cell viability was measured by WST‐1, and the cytotoxic effects of artepillin C on HSC‐3 cells were quantified with flow cytometry. The survivin levels were determined by ELISA.

Results

Artepillin C exhibited dose‐ and time‐dependent cytotoxic effects on HSC‐3 cells. Flow cytometric analysis showed that 22% of untreated HSC‐3 cells underwent spontaneous cell death, whereas 77.32% of the cells were killed in response to the highest dose of artepillin C at 72 hours. Survivin expression was reduced in treated cells.

Conclusions

HSC‐3 cells are vulnerable to artepillin C in a dose‐ and time‐dependent manner. HSC‐3 cell death induced by artepillin C, at least in part, was a result of a decrease in survivin levels.     

Survivin在口腔鳞癌组织中的表达及其与COX-2相关性研究

目的:探讨凋亡抑制基因Survivin在口腔鳞癌组织中的表达及其与COX-2表达的相关性。方法:运用S-P免疫组化技术,检测Survivin和COX-2蛋白在50例口腔鳞癌组织、10例正常口腔黏膜组织中的阳性率。结果:Survivin蛋白在10例正常口腔黏膜组织中呈阴性,而在50例口腔鳞癌组织中有42例阳性,占84%,差异有极显著性差异(P<0.01),COX-2蛋白在口腔鳞癌组织中的阳性率为86%(43/50)。Survivin蛋白阳性率与年龄和性别不相关,而与COX-2蛋白阳性呈正相关(P<0.05)。Survivin蛋白在低分化癌组织中的表达比在高分化癌中高,但在统计学上无明显差异性。结论:肿瘤组织中Survivin蛋白的高阳性表达对口腔鳞癌的发生发展起重要作用;COX-2蛋白在口腔鳞癌组织中也有高阳性表达,并与Survivin有相关性。两者可能存在共同的激活机制,从而抑制口腔鳞癌细胞的凋亡。     

基于免疫相关基因构建口腔鳞癌临床预后模型

目的 筛选口腔鳞状细胞癌(oral squamous cell carcinoma, OSCC)中关键的免疫相关基因(immune-related genes, IRGs),并构建免疫相关预后模型来预测OSCC患者的总生存期,为口腔鳞癌临床治疗方案的制定提供依据。方法 首先从癌症基因组图谱(The Cancer Genome Atlas, TCGA)数据库中下载OSCC基因表达数据和相应临床信息。随后确定差异表达的IRGs,通过单因素Cox回归分析、Lasso-Cox回归分析以及多因素Cox回归分析建立免疫相关预后模型,并验证预后模型的预测性能。结果 本研究构建了由6个IRGs组成的预后模型。低风险组和高风险组的总生存率差异显著(P<0.05)。1、3、5年受试者工作特征(receiver operating characteristic, ROC)曲线的曲线下面积(area under the curve, AUC)分别为0.72、0.73、0.76,这表明预后模型的预测效果较好。随后的单因素和多因素Cox回归分析证明该预后模型的风险评分是独立预后因素(P<0.001)。...     

口腔鳞癌组织中Survivin的表达与血管生成的关系

目的 :观察口腔鳞癌组织中Survivin的表达及其与微血管密度 (microvesseldensity ,MVD)的关系 ,探讨Survivin对口腔鳞癌血管生成的作用及意义。方法 :收集口腔鳞癌标本 42例 ,其中有淋巴结转移者 14例 ,正常口腔黏膜组织 10例 ,用免疫组化染色法探测Survivin和CD3 4 的表达情况并计数微血管密度 (MVD)。结果 :正常口腔黏膜均未见Survivin表达 ,口腔鳞癌中Survivin表达阳性率为 80 .95 % ( 3 4/4 2 )。口腔鳞癌中MVD显著高于正常组织 ,且随病理分化不良而增高 (P <0 .0 5 ) ,有淋巴结转移组MVD显著高于无淋巴结转移组 (P <0 .0 5 ) ,Survivin表达阳性组中MVD明显高于Survivin表达阴性组 (P <0 .0 5 ) ,Survivin的表达与MVD呈正相关 (P <0 .0 5 )。结论 :Survivin在口腔鳞癌组织中的高度表达在口腔鳞癌发生发展中起重要作用 ,并与其血管生成和淋巴结转移有关     

采用RNA干扰技术沉默survivin基因对口腔鳞状细胞癌细胞凋亡的影响

目的 应用RNA干扰（RNAi）技术抑制口腔鳞状细胞癌细胞株HSC-3细胞凋亡抑制因子survivin基因的表达,促进HSC-3细胞的凋亡。方法 合成针对survivin基因的小干扰RNA（siRNA）,转染对数生长期的HSC-3细胞。采用半定量逆转录聚合酶链反应检测HSC-3细胞中survivin mRNA的表达,MTT法测定细胞活性,tunnel分析和流式细胞术检测HSC-3细胞的凋亡,并设阴性对照组和空白对照组。结果 转染siRNA组survivin mRNA的表达明显低于阴性对照组和空白对照组;MTT法测定细胞活性,阴性对照组和空白对照组没有明显差别,但siRNA组的细胞活性明显下降;tunnel分析显示,转染siRNA组的凋亡细胞明显多于阴性对照组和空白对照组;流式细胞术分析,转染siRNA组的凋亡率（24.99％±1.33％）明显高于阴性对照组（1.24％±0.13％）和空白对照组（0.10％±0.02％）。结论 采用siRNA沉默survivin基因能促进口腔鳞状细胞癌细胞的凋亡,survivin siRNA基因治疗有可能成为口腔鳞状细胞癌治疗的新技术。     

Phospholipase C gamma 1 is a potential prognostic biomarker for patients with locally advanced and resectable oral squamous cell carcinoma

The aim of this study was to investigate the prognostic and predictive values of phospholipase C gamma 1 (PLCG1) expression in patients with locally advanced and resectable oral squamous cell carcinoma (OSCC), who were treated in a prospective, randomized, phase 3 trial evaluating standard treatment with surgery and postoperative radiation preceded or not by induction docetaxel, cisplatin, and 5-fluorouracil (TPF). Immunohistochemical staining for PLCG1 was performed on the biopsies of 232 out of 256 OSCC patients at clinical stage III/IVA; the PLCG1 positive score was determined by immunoreactive scoring system. The survival analysis was performed by Kaplan–Meier method; hazard ratios were calculated using the Cox proportional hazards model. Patients with a low PLCG1 expression had a significantly better overall survival (P = 0.022), and a trend towards better disease-free survival (P = 0.087), loco-regional recurrence-free survival (P = 0.058), distant metastasis-free survival (P = 0.053), and a high response rate to TPF induction chemotherapy with regard to clinical response (P = 0.052) and pathological response (P = 0.061), compared to those with high PLCG1 expression. Our results suggest that PLCG1 expression could be used as a prognostic biomarker for patients with advanced OSCC; however, it was not an adequate predictive biomarker for TPF induction chemotherapy.     

外周血嗜酸性粒细胞检测对口腔鳞状细胞癌术后的预后意义

目的探讨外周血嗜酸性粒细胞差异对口腔鳞状细胞癌术后患者的预后意义。方法回顾性分析2010年1月至2012年12月绍兴市人民医院口腔科住院并行手术治疗的口腔鳞状细胞癌患者62例,随访至2014年12月。计算嗜酸性粒细胞百分比及计数的均值,将患者分为大于等于均值组及小于均值组,比较两组患者术后的复发率,并将两组患者进行生存分析及多因素COX回归分析。结果按嗜酸性粒细胞计数均值分组,两组患者复发率及生存率差别均有统计学意义(P值分别为0.002和0.016)。按嗜酸性粒细胞百分比均值分组,两组患者复发率差异有统计学意义(P=0.005),生存率差异无统计学意义(P=0.253)。通过单因素生存分析,嗜酸性粒细胞计数是患者生存率的影响因素(P=0.030),多因素COX回归分析,嗜酸性粒细胞百分比与计数对生存率均无明显影响(P>0.05),而年龄及复发与否是患者生存率的影响因素(P<0.05)。结论口腔鳞状细胞癌术后早期患者外周血嗜酸性粒细胞百分比及计数减少提示该病复发的可能性大;尚无足够证据表明外周血嗜酸性粒细胞的改变将影响患者总体生存率。     

The prognostic significance of histological features in oral squamous cell carcinoma

Background:  Different factors predict nodal metastasis, recurrence and survival in oral cancer. The aim was to assess the prognostic value of histological features related to the primary tumour.
Methods:  A total of 144 patients surgically treated at Odense University Hospital for oral cancer between 1999 and 2004 were included in the study. Postoperative radiation therapy was given in case of close and involved margins or high TNM stages (UICC 1997). Median follow-up time was 38 months. All surgical resections were reviewed and 22 histological characteristics were assessed.
Results:  The predominant sites were floor of mouth (FOM, 39%) and lateral tongue (35%). Fifty-nine per cent had UICC97 stage I–II disease. Five-year cause-specific survival was observed in 65%. Nodal involvement at diagnosis was observed in 36% which was significantly related to grade, neural and vascular invasion; surgical margins and increasing tumour depth. A cut-off value of 2 mm (4 mm for FOM) separated patients without and with nodal metastasis at the time of diagnosis. However, on multivariate analysis, neck disease was only associated with tumour depth and grade. Cox analysis of local recurrence in the oral cavity over time showed that tumour diameter and surgical margins were significant predictors while cause-specific survival was related to diameter, depth of invasion, surgical margins and extracapsular spread (ECS).
Conclusions:  Tumour depth and grade were strong prognostic factors for nodal metastasis, independently of other histological features. Tumour diameter and margins independently predict local recurrences in the oral cavity as well as cause-specific survival. Nodal involvement and ECS were associated with adverse prognosis.     

维吾尔族口腔鳞状细胞癌患者中血清鳞状细胞癌抗原的表达研究

目的 探讨维吾尔族口腔鳞状细胞癌(OSCC)患者血清中鳞状细胞癌抗原(SCC-Ag)的表达.方法 采用化学发光法检测90例维吾尔族OSCC患者(OSCC组)和90例维吾尔族其他口腔恶性肿瘤患者(口腔非OSCC恶性肿瘤组)、101例维吾尔族口腔良性肿瘤患者(良性肿瘤组)和120例维吾尔族正常对照者(正常组)的血清SCC-Ag水平,并对其中60例OSCC和60例口腔非OSCC恶性肿瘤患者治疗前后SCC-Ag水平进行检测,分析SCC-Ag在OSCC诊断、治疗疗效中的意义.结果 OSCC组的SCC-Ag水平明显高于口腔非OSCC恶性肿瘤组(P=0.002),良性肿瘤组(P=0.001)和正常组(P=0.001).OSCC患者血清SCC-Ag阳性率为43.3％(39/90),明显高于其他3组(P=0.000 1).OSCC患者TNM晚期的术前SCC-Ag水平的表达高于早期水平(P<0.05).复发组和未复发组的术前、术后、放化疗后SCC-Ag水平相比较,差异均有统计学意义(P＜0.05).手术及放化疗前后鳞状细胞癌患者血清SCC-Ag水平变化差异有统计学意义(P＜0.0 1),然而,在口腔非OSCC恶性肿瘤组治疗疗效差异无统计学意义(P>0.05).结论 血清SCC-Ag对维吾尔族OSCC患者的早期诊断敏感性不高,但在评价治疗疗效,预测肿瘤复发方面有重要的临床意义.     

Evaluation of myofibroblasts in oral epithelial dysplasia and squamous cell carcinoma

Background:  Carcinogenesis is accompanied by a number of changes in the adjacent stroma including the appearance of myofibroblasts. The purpose of this study was to evaluate and compare the presence of myofibroblasts in normal mucosa, oral epithelial dysplasia, and different grades of oral squamous cell carcinoma.
Methods:  The study sample consisted of three groups, including 40 oral squamous cell carcinomas, 15 dysplasias, and 15 sections of normal oral epithelium. Vimentin, desmin, and alpha-smooth muscle actin were used to identify myofibroblasts.
Results:  The percentage and intensity of alpha-smooth muscle actin were examined, and positive immunostaining was observed in the myofibroblasts of all squamous cell carcinomas; however these cells did not stain in the dysplasias or normal epithelium specimens. The presence of myofibroblasts was significantly higher in oral squamous cell carcinomas compared to both, dysplasias and normal mucosa cases ( P  < 0.001). A significant difference was not observed between the different grades of oral squamous cell carcinoma ( P  = 0.2).
Conclusions:  These findings show the presence of myofibroblasts in the stroma of oral squamous cell carcinoma but not dysplasia and normal mucosa, suggesting further investigation to clarify the role of myofibroblasts in the carcinogenesis of this tumor.     

The prevalence of oral leukoplakia in 138 patients with oral squamous cell carcinoma

OBJECTIVES: To determine the relationship between oral leukoplakia (OL) and oral squamous cell carcinoma (OSCC), and to evaluate possible differences between those carcinomas with and without associated leukoplakia. MATERIAL AND METHODS: A total of 138 patients were studied at the Stomatology Service of the University General Hospital, Valencia, Spain. These patients were divided into two groups: group 1, patients with oral cancer and leukoplakia, and group 2, patients with OSCC but with no associated premalignant lesions. The relationship between this precancerous lesion and the OSCC was evaluated, as well as the possible clinical and histological differences between the tumours of the two groups. RESULTS: Leukoplakia was detected in 27 (19.56%) patients with OSCC. No differences were found between the two groups regarding age and tumour location. However, statistically significant differences were observed with respect to the form, tumour stage and the presence of adenopathies in the cancers with and without leukoplakia; in that the tumours associated with leukoplakia were diagnosed as being at a more initial stage. CONCLUSIONS: Those patients with OL associated with oral cancer presented with tumours at a less advanced stage than those where no associated leukoplakia existed.