NKG2D及其配体MICA/B在口腔鳞癌患者外周血NKT细胞中的表达及其意义

目的:探讨NKG2D及其配体在口腔鳞癌患者与正常人中外周血NKT的表达及其临床意义。方法:通过流式细胞术检测28例口腔鳞癌及25例正常人的NKT细胞计数,PCR检测NKG2D及其配体在不同组中NKT及配体MICA/B的表达情况,分析表达率与临床意义。结果:与正常组相比口腔鳞癌病人中的外周血中CD3+CD56+NKT的含量较低((1.74±0.15)个/μL),并且口腔鳞癌患者CD3+CD56+NKT细胞中NKG2D,MICA和MICB表达均下降,与对照组比较差异有统计学意义。结论:口腔鳞癌患者外周血中的NKT细胞数量和NKG2D及其配体MICA/B表达量相对下降,这可能影响NKT在肿瘤免疫中的作用,使口腔鳞癌细胞逃避免疫监视。他们可以作为口腔鳞癌免疫状态的参考指标,也为口腔鳞癌的治疗提供一种新的方向。     

EGCG对舌鳞癌Tca8113细胞增殖与凋亡的实验研究

目的:研究表没食子儿茶素没食子酸酯(EGCG)对体外培养的人舌鳞癌Tca8113细胞增殖抑制及诱导凋亡的作用。方法:采用CCK-8法检测不同浓度的EGCG(0、10、20、40、80mg/L)对Tca8113细胞的体外增殖抑制作用;采用流式细胞术及DAPI荧光染色法检测EGCG对Tca8113细胞凋亡的影响。结果:CCK-8法检测显示EGCG对Tca8113细胞有明显增殖抑制作用并呈现时间及浓度依赖性(P<0.01);流式细胞仪结果显示,经10mg/L EGCG处理的Tca8113细胞凋亡率还不能认为与对照组有区别,其余EGCG处理组细胞凋亡率均高于对照组并呈现时间及浓度依赖性(P<0.01);DAPI染色在免疫荧光显微镜下可见典型的细胞核皱缩及凋亡小体等。结论:EGCG对Tca8113细胞具有明显的增殖抑制及诱导凋亡作用,且呈现时间及浓度依赖性。     

Is panendoscopy a necessary staging procedure in patients with lacking risk factors and oral squamous cell carcinoma?

ObjectivesRoutine panendoscopy is used to detect synchronous malignancies of the upper aerodigestive tract in staging of oral squamous cell carcinoma. The goal of this study was to investigate the occurrence of synchronous malignancies at time of diagnosis using panendoscopy. To challenge the role of panendoscopy as inherent part of routine staging procedures, we were especially interested in low risk patients.Materials and methodsRetrospectively, a cohort of 484 patients with pathologically confirmed diagnosis of primary oral and oropharyngeal squamous cell carcinoma was investigated. Electronically recorded findings of in-house conducted panendoscopy were retrieved and evaluated for the occurrence of pathological changes of the mucosa. In case of synchronous malignancies, findings were correlated to preoperative radiographic imaging. Patients were classified as high or low risk. Patients with lacking risk factors (no smoking, no drinking in history) were defined as low risk patients.ResultsOverall, we detected three synchronous malignancies of the upper aerodigestive tract (3/484; 0.6%). Two non-small cell lung cancers were detected in patients with a smoking history of 60 pack years. One esophageal carcinoma in situ was detected in a patient with reported alcohol consumption. No synchronous malignancy was detected in patients without risk factors and no malignancy was previously detected by diagnostic imaging.ConclusionPre-treatment panendoscopy can reveal synchronous malignancies of the upper aerodigestive tract in patients with primary oral squamous cell carcinoma. Risk stratification of patients can avoid unnecessarily conducted panendoscopy in patients without risk factors. This may lead to a higher cost-efficacy in public health system, less treatment-related complications and earlier treatment initiation.     

口腔鳞癌发展过程中抑癌基因PTEN的蛋白表达及意义

目的：探讨抑癌基因PTEN(phosphatase and tensin homologue deleted on chromosome ten)在口腔鳞癌(oral squamous cell cancer，OSCC)发展过程中的蛋白表达及临床病理意义。方法：应用免役组化S—P法检测10例正常口腔粘膜、10例上皮单纯增生、15例上皮异常增生及32例OSCC组织中抑癌基因PTEN的蛋白表达情况，同时结合患者的临床病理资料进行分析。结果：正常口腔粘膜和上皮单纯增生组织中VTEN蛋白全部阳性表达；上皮异常增生组织中PTEN蛋白阳性表达率为93．3％；OSCC组织中PTEN蛋白的阳性表达率为71．9％，其中高、中、低分化OSCC组织中PTEN蛋白的阳性表达率分别为85．7％、75％和33．3％。PTEN在OSCC组织中的蛋白表达与患者的性别、年龄无明显相关性，但与组织分化程度明显相关。结论：OSCC组织(尤其低分化)中PTEN蛋白的阴性表达率较高．说明PTEN基因突变或缺失在OSCC的发生、发展过程中起重要作用。     

Multiplexed immunobead‐based assay for detection of oral cancer protein biomarkers in saliva

Objective: For clinical applications of biomarkers, there is a need for multiplex assays using high throughput platforms. The objective of this study was to determine the efficacy of Luminex Multianalyte Profiling (xMAP) technology for measurement of salivary proteins and to evaluate whether multiplex assays are as effective as single‐plex assays and enzyme‐linked immunosorbent assay (ELISA). Results: The average levels of interleukin‐8 (IL‐8) from the single‐plex assay were 3313.2 ± 3759.8 pg ml^?1 [oral squamous cell carcinoma (OSCC), n = 20] and 1061.7 ± 1978.8 pg ml^?1 (control, n = 20). The IL‐1β average levels from the single‐plex assay were 945.2 ± 1134.8 pg ml^?1 (OSCC, n = 20) and 314.2 ± 444.8 pg ml^?1 (control, n = 20). The average levels of IL‐8 from the multiplex assay were 2834.9 ± 3385.6 pg ml^?1 (OSCC, n = 20) and 947.3 ± 2036.8 pg ml^?1 (control, n = 20). The IL‐1β average levels from the multiplex assay were 1013.5 ± 1221.1 pg ml^?1 (OSCC, n = 20) and 376.3 ± 576.3 pg ml^?1 (control, n = 20). The correlation coefficient between Luminex and ELISA assay for IL‐8 (n = 19) and IL‐1β (n = 19) was 0.91 and 0.84, respectively. Conclusion: Luminex xMAP single‐plex and multiplex assays are as effective as ELISA assays for quantification of proteins in saliva. Both IL‐8 and IL‐1β were expressed at significantly higher levels in OSCC subjects than in the matched healthy control subjects.     

苯丙芘联合佛波酯恶性转化HIOEC细胞的实验研究

目的:诱导永生化口腔上皮细胞系HIOEC细胞恶性转化,建立HIOEC-B(a)P-TPA细胞系。方法:通过递增浓度的化学致癌剂苯丙芘复合促癌剂佛波酯体外诱导HIOEC细胞,筛选得到具有恶性表型的鳞状细胞癌细胞系HIOEC-B(a)P-TPA。通过微分干涉显微镜和HE染色观察细胞形态学改变,免疫组化染色检测细胞角蛋白和波形蛋白表达情况;软琼脂集落形成、裸小鼠异体皮下成瘤性鉴定HIOEC-B(a)P-TPA细胞的恶性表型。结果:(1)HIOEC细胞诱导培养6个月后,可以在正常生理钙离子浓度、含胎牛血清培养液中生长;(2)细胞在诱导过程中形态多变,最后以成纤维样细胞为主,异形性明显;(3)HIOEC-B(a)P-TPA细胞中细胞角蛋白表达减弱,同时表达波形蛋白;(4)有很强的软琼脂集落形成能力,集落形成率为24.5%;(5)HIOEC-B(a)P-TPA细胞尚未观察到成瘤性。结论:生物化学致癌因素B(a)P和TPA可诱导HIOEC细胞恶性转化,反映了口腔黏膜上皮细胞体外恶变发生、发展过程。     

MDC1蛋白在口腔鳞状细胞癌中的表达及意义

目的：探讨口腔鳞状细胞癌中MDC1（mediator of DNA damage cheekpoint protein1）蛋白的表达水平及意义。方法：采用免疫组织化学Envision法检测10例正常口腔黏膜、60例口腔鳞状细胞癌中MDC1的表达水平．并分析其与临床病理参数之间的关系。结果：MDC1蛋白在口腔鳞状细胞癌中表达高于正常口腔黏膜，两者之间差异有统计学意义（P=0．042），而且MDC1蛋白的表达升高与淋巴结转移有关（P=0．033），但与病理分级（P=0．986）、患者的年龄（P=0．723）、性别（P=0．142）无关。结论：MDC1蛋白在口腔鳞状细胞癌中表达增高，提示它可能在肿瘤的发生和发展中具有一定的作用，为进一步理解细胞DNA损伤与肿瘤发生的关系提供了依据。