多重PCR对头颈部肿瘤中MTS_1基因外显子2的检测分析

目的　为了解MTS1基因变异在头颈部肿瘤发生发展中的作用。方法　采用多重PCR技术扩增MTS1基因外显子 2。结果　本组MTS1基因外显子 2纯合子缺失的频率为 37.3 % (17/4 5 ) ,分化较好鳞癌MTS1基因缺失频率 31.5 % (6 /19)明显低于分化较差的鳞癌的 5 7.1% (8/14) ,但差异无显著性 (P >0 .0 5 )。结论　本研究提示MTS1基因变异与头颈部恶性肿瘤的发生发展有关     

口腔鳞癌根治性手术前后血清TGF-β1的变化

目的　探讨转化生长因子β1 (TGF-β1 )和口腔鳞癌侵袭及转移的关系。方法　应用酶联免疫吸附法测定 40例口腔鳞癌根治性手术前后患者血清 TGF- β1 浓度 ;免疫组化法分析癌灶组织的 TGF- β1 蛋白表达水平。结果　伴有颈淋巴结转移组的血清 TGF- β1 浓度和 TGF- β1 蛋白表达水平高于无淋巴结转移组 (P<0 .0 5～ 0 .0 1) ,切除癌灶后上述各组的血清 TGF- β1 浓度均明显下降 (P<0 .0 5 )。结论　 TGF- β1 可能与人口腔鳞癌的侵袭和转移有密切关系     

口腔颌面部鳞癌组织中p16基因变异的初步研究

目的 检测口腔颌面部鳞癌中p16基因的缺失和突变，以了解p16基因在口腔颌面部恶性肿瘤发生发展过程中的作用。方法 应用聚合酶链反应－单链构象多态性分析（PCR－SSCP）技术，检测33例原发性口腔颌面部鳞癌的石蜡标本，将p16基因的变异频率与口腔颌面部鳞癌的临床分期，组织学分级及淋巴结转移进行统计学分析。结果 p16基因的缺失率为21％，点突变率为6％，变异频率为27％。临床Ⅰ、Ⅱ、Ⅲ、Ⅳ期每两期之间无显著性差异（P＞0．05）；而临床Ⅰ、Ⅱ期和Ⅲ、Ⅳ期之间有显著性差异（P＜0．05）。组织学分级和有，无淋巴结转移之间p16基因变异无显著性差异（P＞0．05）。结论 p16基因的缺失和突变是口腔颌面部鳞癌中常见的分子事件，它的失活在口腔颌面部鳞癌的发生发展中起着重要作用，说明其变异与肿瘤的临床分期密切相关，随着肿瘤临床进程的发展，其变异频率增高。研究未发现其变异与肿瘤组织学分级和淋巴结转移之间存在相关关系。     

FHIT,Livin,Smac在口腔鳞癌中的表达及意义

目的:探讨FHIT、Livin、Smac蛋白在口腔鳞癌发生发展中的作用及其相互关系.方法:采用免疫组化SP法检测9例口腔正常黏膜上皮,17例口腔癌前病变,38例口腔鳞癌,10例淋巴结转移灶中FHIT、Livin、Smac的表达.结果:FHIT在口腔正常组、癌前病变组、鳞癌组、淋巴结转移灶组中的阳性率分别为:100.0%、100.0%、76.3%、50.0%,正常组与鳞癌组,癌前病变组与鳞癌组,表达的差异有显著性意义.Livin在口腔正常组、癌前病变组、鳞癌组、淋巴结转移灶组中的阳性率分别为55.6%、58.8%、100.0%、100.0%,正常组与鳞癌组,癌前病变组与鳞癌组,鳞癌组中未转移与有转移,表达的差异有显著性意义.Smac在口腔正常组、癌前病变组、鳞癌组、淋巴结转移灶组中的阳性率分别为100.0%、100.0%、81.6%、60.0%,正常组、癌前病变组合并后与鳞癌组表达的差异有显著性意义.结论:FHIT或/和Smac蛋白缺失可能是口腔鳞癌发生过程中的早期表现;Livin蛋白的异常表达可能对口腔鳞癌的发展起作用;在口腔鳞癌中存在FHIT、Livin和Smac基因的相互调节.     

EMS1基因扩增与口腔黏膜癌变的相关研究

目的 了解EMS1基因扩增是否参与口腔黏膜癌变。方法采用显微解剖技术分别获取正常口腔黏膜上皮，口腔白斑患者的单纯增生，轻、中、重度异常增生上皮和原发性口腔鳞癌组织标本78例，应用差示PCR反应检测EMS1基因扩增。结果①分别有20．0％的口腔白斑组织，57．6％的口腔鳞癌组织观察到EMS1扩增；②在口腔黏膜癌变进程中，EMS1扩增开始于中度异常增生黏膜，在伴有淋巴结转移的口腔鳞癌组织中其扩增率有显著增高(P=0．015)。结论EMS1基因扩增与口腔黏膜癌变的演进相平行，似是口腔黏膜癌变的早期分子事件。     

口腔鳞状细胞癌组织中P16蛋白的免疫组化定量分析

目的　探讨P16蛋白在口腔鳞状细胞癌(鳞癌)组织中的表达及与口腔鳞癌发生发展的关系。方法　采用免疫组化S -P法,对5 6例口腔鳞癌组织、30例癌旁组织及10例正常口腔黏膜中P16蛋白的表达进行检测,应用全自动图像分析仪对染色结果进行定量测定。结果　口腔鳞癌组织中P16蛋白的表达量低于癌旁组织及正常黏膜(P <0 .0 5 ) ,口腔鳞癌Ⅲ级低于Ⅰ、Ⅱ级(P <0 .0 5 )。颈淋巴结转移组低于无颈淋巴结转移组(P <0 .0 5 )。结论　P16蛋白在口腔鳞癌组织中的表达常为缺失,说明其作为抑癌基因在口腔鳞癌的发生中起作用。P16蛋白的表达程度越低,分化越差,肿瘤越易于浸润和转移。     

nm23-H1基因在口腔鳞癌颈淋巴结转移不同阶段的调控作用

目的　探讨nm23-H1基因在口腔鳞癌区域淋巴结转移不同阶段的调控作用及影响。方法　利用免疫组化和Western Blot技术,结合淋巴结转移和原发灶浸润分型的资料,分组对照研究nm23-H1基因在200例口腔鳞癌颈淋巴结转移不同阶段的表达情况。结果　淋巴结转移组nm23-H1基因阴性表达率明显高于无淋巴结转移组(P<0·01)。N1期、转移仅累及1个淋巴结、Ⅳc型浸润,以及低分化肿瘤的患者,均有比其它患者明显增高的 nm23-H1基因的阴性表达率(P<0·01);转移至颌下或颌下-颈深上淋巴结平面者的阴性表达率,也高于转移到其它平面者(P<0·05)。结论　nm23-H1基因主要是通过干扰肿瘤实体内高转移细胞亚群的形成和初始转移的发生来调控口腔鳞癌患者的颈淋巴结转移。     

口腔鳞癌中促血管生成素-2的表达与临床病理学特征及血管生成的关系

目的 :研究口腔鳞癌中促血管生成素 2 (Angiopoietin 2 ,Ang 2 )的表达及其与临床病理学特征和微血管密度 (microvesseldensity ,MVD)间的关系。 方法 :用免疫组化SABC法检测口腔鳞癌标本 41例 (有淋巴结转移者13例 )及 3 0例癌旁正常组织和 10例正常口腔黏膜组织中的Ang 2和CD3 4表达并计数微血管密度 (MVD )。结果 :Ang 2表达主要定位于肿瘤细胞胞浆 ;Ang 2在口腔鳞癌组织中的表达显著高于癌旁正常组织 (P <0 .0 5 )和正常口腔黏膜组织 (P <0 .0 1) ;Ang 2表达与肿瘤的淋巴结转移密切相关 (P <0 .0 1) ,而与病人的性别、年龄和TNM分期及肿瘤分化程度无关 (P >0 .0 5 ) ;Ang 2表达阳性组中MVD显著高于Ang 2表达阴性组 (P <0 .0 5 ) ;Ang 2的表达与MVD呈正相关 (P <0 .0 1)。结论 :Ang 2在口腔鳞癌组织中的高表达可能在口腔鳞癌的发展中起重要作用 ,并与肿瘤的淋巴结转移和血管生成有关。     

艾滋病感染者口腔疣状肿块人类乳头状瘤病毒感染与P53,Ki-67蛋白过度表达

目的 :了解艾滋病感染者口腔黏膜疣状肿块细胞生物学特性。方法 :应用免疫组织化学法、PCR对艾滋病感染者口腔黏膜疣状肿块、非艾滋病感染者口腔黏膜癌前病变和鳞癌组织中P5 3和Ki 67蛋白、人类乳头状瘤病毒 (HPV ) ,巨细胞病毒 (CMV)和EB病毒 (EBV )进行检测。结果 :( 1)艾滋病感染者口腔黏膜疣状肿块中P5 3蛋白阳性表达率为 2 3 % ,Ki 67蛋白阳性表达率为 76% ,二者均低于非艾滋病感染者口腔黏膜鳞癌(P <0 .0 5 ) ,但与非艾滋病感染者口腔黏膜癌前病变无明显差别 (P >0 .0 5 ) ;( 2 )艾滋病感染者口腔黏膜疣状肿块中 ,HPV感染率为 88.2 % ,明显高与非艾滋病感染者口腔黏膜癌前病变和口腔黏膜鳞癌 (P <0 .0 1)。没有检测到EBV、CMV病毒感染。结论 :艾滋病感染者口腔疣状肿块和HPV感染有关 ,存在抑癌基因突变和细胞过度增殖现象。     

诱导型一氧化氮合酶在口腔黏膜癌前病变——白斑中的表达及意义

目的　探讨口腔黏膜癌前病变 (口腔白斑 )及口腔鳞癌的发生、发展过程中诱导型一氧化氮合酶 (iNOS)的表达及意义。方法　应用免疫组化法对 10例正常口腔黏膜、30例口腔白斑、10例口腔鳞癌分别检测iNOS的表达。结果　iNOS在正常口腔黏膜、口腔白斑、口腔鳞癌的表达阳性率分别为 0、6 6 .7%、80 .0 % ,各组间差异均有非常显著性 (P <0 .0 1)。结论　由iNOS诱导产生的一氧化氮 (NO)可能在口腔鳞癌的发生、发展中起着重要作用。     

Alterations of p16/MTS1 gene in oral squamous cell carcinomas from Taiwanese

To determine the alterations of the p16/MTS1 gene in oral squamous cell carcinoma (OSCC), we examined in Taiwanese patients the mutation, deletion and methylation of p16/MTS1 in primary OSCCs associated mostly with betel quid (BQ)/tobacco use. Among 110 tumors undergoing mutational analyses, seven (6%) showed mutations in exon 2 or the intron 1/exon 2 splice site. All but one mutation disrupted the encoded proteins. Base transitions represented the vast majority (6/7) of the mutations identified in BQ/tobacco consuming subjects. It was noted that 15/56 (27%) tumors examined by restriction fragment methylation analysis revealed a significant level of methylation in different loci of exon 1 as compared with the respective non-cancerous tissue. Mutation of p16/MTS1 was exclusively identified in carcinomas of buccal mucosa, whereas methylation of the p16/MTS1 promoter region occurred preferentially in carcinomas of the tongue (54%) rather than at other sites (22%). Homozygous deletion was not found in 56 paired samples examined, nor was hemizygous deletion indicated in 12 informative cases. The results indicated aberrant methylation and mutation as the molecular abnormality of p16/MTS1 in the OSCC from Taiwanese.     

MTS1 gene mutations in archival oral squamous cell carcinomas

Multiple tumor suppressor gene 1 (MTS1) has been found mutated or deleted in a variety of human cancers. Our purpose was to identify and characterize MTS1 gene mutations in primary oral squamous cell carcinomas (SCCs) in each of the three exons of the MTS1 gene. Seventeen archival samples of oral SCC were evaluated for the presence of MTS1 mutations using single strand conformation polymorphism (SSCP) and DNA sequencing. Three of 17 tumors exhibited MTS1 gene mutations; one tumor exhibited a mutation in exon 2 and two tumors exhibited mutations at the splice site junction of intron 2 and exon 3. Three tumors also exhibited a common base change in the 3'untranslated region of exon 3, which is interpreted as a likely polymorphic variant. An examination of the three tumors exhibiting MTS1 point mutations revealed no unique characteristics relative to p53 immunohistochemical activity, mitotic frequency, or degree of histologic differentiation. This study indicates that MTS1 gene mutations may be involved in at least a minor proportion of oral SCCs.     

环氧酶-2在人舌鳞癌及癌前病变组织中的表达

目的　探讨环氧酶 2 (COX 2 )在口腔癌前病变和舌癌组织中的表达及其意义。方法　采用免疫组化法(IHC)检测 6 0例人舌鳞状细胞癌和 2 0例癌前病变石蜡标本的COX 2表达。结果　COX 2在正常舌粘膜上皮组织中仅有 1例 (1/ 10 )弱表达 ,在舌粘膜上皮异常增生及舌癌组织中的阳性率分别为 6 5 0 %、78 3% ,舌癌组织中的表达强度明显高于舌粘膜上皮异常增生组织 (P =0 0 2 17)。结论　COX 2在舌癌及其癌前病变中有不同程度的高表达 ,可以作为化学预防和治疗的靶点     

口腔粘膜鳞癌和癌前病变p53、ras基因突变的检测及p53、p21蛋白的表达

目的　探讨 p5 3、ras基因改变在口腔粘膜鳞癌及癌前病变发生过程中的意义。方法　应用聚合酶链反应和免疫组织化学法检测 2 3例口腔粘膜鳞癌和 15例癌前病变 p5 3、ras基因突变和 p5 3、p2 1蛋白的表达。结果　43.5 % (10 / 2 3)的口腔粘膜鳞癌存在 p5 3基因第 8外显子突变 ,47.8% (11/ 2 3) ras基因有突变 ;口腔粘膜鳞癌 p5 3、p2 1蛋白表达阳性率分别为 43.5 % (10 / 2 3)和 6 0 .9% (14/ 2 3)。癌前病变有 1例存在 p5 3基因第 8外显子和 ras基因突变 (6 .7% ) ;p5 3、p2 1蛋白表达阳性率分别为 13.3% (2 / 15 )和 2 0 % (3/ 15 )。两组间差异显著 (P<0 .0 5 )。结论　 p5 3、ras基因突变和 p5 3、p2 1蛋白过量表达与口腔粘膜鳞癌的发生过程密切相关 ,可作为癌前病变发展为癌危险性的生物指标     

口腔鳞癌和癌前病变中端粒酶活性的检测

目的 探讨端粒酶活化在口腔粘膜鳞癌、癌前病变中的表达状况以及癌变过程中的作用。方法 采用端粒酶（ｔｅｌｏｍｅｒａｓｅ）聚合酶链反应（ｐｏｌｙｍｅｒａｓｅ ｃｈａｉｎ ｒｅａｃｔｉｏｎ,ＰＣＲ）、酶联免疫吸附法和端粒重复放大（ｔｅｌｏｍｅｒｅ ｒｅｐｅａｔ ａｍｐｌｉｆｉｃａｔｉｏｎ ｐｒｏｔｏｃｏｌ,ＴＲＡＰ）电泳法对２０例口腔粘膜鳞癌、１０例口腔粘膜白斑、２０例口腔粘膜扁平苔藓和１０例正常口腔粘     

The correlation between alteration of p16 gene and clinical status in oral squamous cell carcinoma

The purpose of the study was to evaluate the presence of alteration of the tumor suppressor gene p16 and to correlate these changes with the clinical status of the patients in oral squamous cell carcinoma. Forty-eight oral squamous cell carcinomas were included in the analyses. Deletion analysis was performed by the polymerase chain reaction (PCR). Mutation analysis was restricted to exon 1 and exon 2 of the p16 gene, previously shown to have a high incidence of mutations. The sequences containing exon 1 and exon 2 were amplified by PCR and screened with a single-strand conformation polymorphism (SSCP) technique. Samples showing band shifts in SSCP were sequenced by PCR direct sequencing. Western blots were used to detect the protein expression of the p16 gene, and the results were evaluated with regard to their biological relevance in correlation with clinicopathological factors. Seven (14.6%) deletions were found; 5 (10.4%) mutations were discovered and located in different codons; 26 (54%) specimens had no p16 protein expression; in 11 specimens with p16 deletion or mutation, p16 protein could not be detected. One mutation was non-sense. The p16 gene alterations showed no relationship with location and clinical stage of cancer; however, a close relationship between p16 alterations and cancer metastasis to neck lymph node was found. The alteration rate gradually elevated from well to poorly differentiated grades. We perceive two results. First, the alterations of the p16 gene are common in oral squamous cell carcinoma. Second, the alterations of the p16 gene may attribute to the metastatic behavior or histological grade of cancer cells.     

Nachweis von TP53-Mutationen mittels Exfoliativzytologie (brush biopsy) oraler Leukoplakien

Objective. The aim of this study was to determine the prevalence of mutations of the tumour suppressor gene TP53 in oral leukoplakias. Material and method. Brush biopsy specimens of 43 oral leukoplakias, 26 oral squamous cell carcinomas (OSCC) for reference, and the oral mucosa of 4 clinically normal volunteers were collected. DNA of the critical exons 5–8 was analysed by temperature gradient gel electrophoresis (TGGE). Results. The prevalence of mutations was 57.7% in OSCC, 39.5% in leukoplakias and 0% in controls. The highest frequency of mutations was found in exon 5 (46.2%) in OSCC and in exon 6 (23.3%) in leukoplakia. More than one mutation was detected in 26.9% of OSCC and 7% of leukoplakia specimens. At least one mutation was found in 37.5% of T1 OSCC and 100% of T4 OSCC specimens and in 37.1% of the L1 leukoplakia and 100% of L3 leukoplakia specimens. Conclusions. TP53 mutations could be a useful prognostic indicator in precancerous oral lesions. Although the brush biopsy technique appears simple clinically, further investigations are necessary to specify the implications of genetic analysis.     

端粒酶hTR基因在口腔癌前病变及鳞癌中的表达

目的探讨端粒酶ｈＴＲ基因在口腔癌前病变及鳞癌中的表达。方法用原位杂交技术检测８２例标本,其中正常口腔粘膜７例,上皮单纯性增生７例,上皮异常增生３０例,原位癌、鳞癌３８例。结果正常口腔粘膜及上皮单纯性增生组织中ｈＴＲ表达较弱,阳性细胞主要位于基底层,检出率２８．５６％（４／１４）。癌前病变中随着异常增生程度的增加,ｈＴＲ表达逐渐增强,阳性细胞逐步由基底层向棘层波及,检出率６０％（１８／３０）。原位癌及鳞癌均有较强的ｈＴＲ表达,检出率为８１．５８％（３１／３８）。结论端粒酶的激活可能出现在口腔癌前病变的晚期阶段,在口腔鳞癌的形成过程中起了关键性作用。     

C‐deletion mutation of the p53 gene at exon 4 of codon 63 in the saliva of oral squamous cell carcinoma in central India: a preliminary study

Aim: The purpose of this study was to detect the C‐deletion mutation of the p53 gene at exon 4 of codon 63 in the saliva of oral squamous cell carcinoma in central India. Methods: The study was carried out in 30 oral squamous cell carcinoma cases and five healthy controls with no habit of betel nut and tobacco chewing. The C‐deletion mutation of the p53 gene at exon 4 of codon 63 was detected in the saliva samples by using polymerase chain reaction. Results: In this study, C‐deletion at exon 4 of codon 63 was detected in 28 of 30 oral squamous cell carcinoma cases (93.33%), but was negative in all five healthy controls and two oral squamous cell carcinoma cases. Conclusions: The results indicated that C‐deletion mutation at exon 4 of codon 63 of the p53 gene in the saliva might be a plausible molecular marker for oral squamous cell carcinoma patients with a habit of betel nut and tobacco lime quid chewing. The results further emphasize the presence of p53 gene mutation in patients with oral squamous cell carcinoma, which can be detected in the saliva through polymerase chain reaction.     

端粒酶蛋白催化亚基(hTRT)mRNA在口腔鳞癌形成过程中的表达

目的:观察端粒酶催化蛋白基因hTRT在口腔粘膜恶性转化不同阶段中的表达,探讨其与口腔粘膜细胞恶性程度的关系。方法:用原位杂交技术检测82例标本,其中正常口腔粘膜7例、上皮单纯性增生7例、上皮异常增生30例、原位癌8例、口腔粘膜鳞癌30例。结果:正常口腔粘膜、上皮单纯性增生组织中hTRT的mRNA表达较弱,阳性信号仅局限于上皮基底层及副基底层间,阳性率21.4%(3/14);上皮异常增生中hTRTmRNA阳性表达见于多层上皮细胞,并随细胞异形性增高而表达增强,阳性率46.7%(14/30);口腔粘膜鳞癌组织中hTRT的mRNA有较强阳性表达,阳性率81.6%(31/38)。结论:端粒酶hTRT基因的表达与口腔粘膜细胞的恶性程度密切相关,端粒酶的重新激活在口腔鳞癌的形成过程中起了关键性作用