涎腺良恶性多形性腺瘤中癌基因C—myc表达的斑点杂交研究

目的 探讨癌基因Ｃ－ｍｙｃ的改变与良恶性多形性腺瘤发生的关系。方法 采用斑点杂交研究４２例涎腺良恶性多形性腺瘤中Ｃ－ｍｙｃ癌基因的表达。结果 涎腺多形性腺瘤中Ｃ－ｍｙｃ癌基因表达均值高于正常腮腺组织（Ｐ〈０．０５）。结论 在涎腺良恶性多形性腺瘤的发生过程中,Ｃ－ｍｙｃ癌基因有激活和过量表达。     

涎腺肿瘤与癌基因c—myc抑癌基因p53和人类乳头状病毒感染的 …

本实验采用免疫组化、ＰＣＲ技术和核酸电泳方法，对涎腺多形性癌和其它类型良恶性进行对比研究。结果显示：在涎腺多形性腺瘤中，ｃ－ｍｙｃ中和Ｐ５３阳生率５０％和２５％，均远低于涎腺恶性肿瘤阳性率１００％，提示ｃ－ｍｙｃ表达与涎腺上皮细胞增殖速率加快有关，其表达率升高，与细胞癌变有密切联系。Ｐ５３阳性表达率虽在恶性肿瘤较高，但仅为判断细胞分化程度的一个参考指标。关于腮腺肿瘤一ＨＰＶ关系，１５例多形性腺瘤与     

野生型p53对涎腺多形性腺瘤细胞突变基因的修复

目的检测涎腺多形性腺瘤基因突变方式及野生型p53对突变基因的修复。方法留取4例涎腺多形性腺瘤新鲜标本，进行原代细胞培养。采用野生型p53基因转染培养细胞，通过逆转录聚合酶链反应检测转染后的基因表达。提取转染后各例细胞DNA及对应的肿瘤组织DNA，分别进行PCR反应扩增，聚丙烯酰胺凝胶电泳及核酸序列测定分析。结果PCR-单链构象多态性分析显示，3例出现异常电泳带。核酸序列测定结果显示，第6外显子的第203号密码子点突变；第8外显子的第272至290号密码子之间出现碱基缺失和碱基插入。野生型p53转染后的细胞DNA序列分析显示，p53第6和第8外显子的5个突变位点均恢复正常。结论涎腺多形性腺瘤发生过程中具有较高频率的p53基因突变，突变方式多样，涉及多个密码子；外源性野生型p53可有效地修复涎腺多形性腺瘤细胞中突变的p53基因位点。     

涎腺多形性腺瘤p53基因突变的聚合酶链式反应--单股构型多态分析研究

目的　检测涎腺多形性腺瘤中 p5 3基因突变。 方法　采用非同位素聚合酶链式反应———单股构型多态分析 (PCR -SSCP)技术对涎腺多形性腺瘤进行研究。结果　 14例多形性腺瘤中有 11例出现PCR -SSCP异常电泳带 (阳性率 78.6 % )。突变部位分布在第 5～ 8外显子 ,其中第 5、6、7、8外显子的阳性例数分别为 2、6、7、3例。在 11例阳性肿瘤中有 2例肿瘤同时出现 3对外显子 (第 6、7、8外显子 )异常 ;3例肿瘤同时出现 2对外显子(第 6、7和 7、8外显子 )异常。结论　①涎腺多形性腺瘤发生中 p5 3基因的第 5～ 8外显子均有突变 ,其中突变频率最高的是外显子 6和 7;②在阳性的肿瘤中约有半数出现多位点突变。     

S-100蛋白和中间丝在涎腺多形性腺瘤中的表达

目的 观察Ｓ－１００Ａ１、Ｓ－１００Ａ４、Ｓ－１００Ａ６、Ｓ－１００Ｂ、Ｋ８．１２、ＫＬ１、波形蛋白（Ｖｉｍｅｎｔｉｎ）、胶质纤维酸性蛋白（ＧＦＡＰ）和神经元特异性烯醇化酶（ＮＳＥ）在涎腺多形性腺瘤中的表达，探讨Ｓ－１００蛋白新亚型的分布规律和作用机制以及肿瘤性肌上皮细胞的生物学特性。方法 将２３例正常涎腺和６０例涎腺多形性腺瘤常规石蜡包埋，制成４μｍ厚的连续切片，行ＨＥ和免疫组化染色。结果 在涎     

c—erbB—2的表达与涎腺恶性多表性腺瘤预后的关系

本研究的目的在于观察癌基因ｃ－ｅｒｂＢ－２表达与涎腺恶性多形性腺瘤患者预后的关系。采用抗ｃ－ｅｒｂＢ－２癌蛋白单克隆抗体ＡＢ－３，应用免疫组织化学方法，对６０例涎腺恶性多形性腺瘤中心ｃ－ｅｒｂＢ－２癌基因表达产物进行了检测。结果发现，２５例ＭＰＡＪｃ－ｅｒｂＢ－２表达产物阳性，阳性表达率４１．６７％。结合临床随访资料分析，ＭＰＡ复发组ｃ－ｅｒｂＢ－２阳性表达率明显高于未复发组（Ｐ＜）．０５）；ｃ－     

癌基因c-myc抑癌基因p53在涎腺肿瘤中的表达

方法：本文采用免疫组化（ＳＰ）法，研究了３０例涎腺肿瘤。目的：观察ｃ－ｍｙｃ癌基因和ｐ５３抑癌基因在涎腺肿瘤中的表达。结果：多形性腺瘤中，ｃ－ｍｙｃ和ｐ５３阳性率分别为５０．０％和２５．０％，均分别低于恶性肿瘤的阳性率７５％和５０．８％，两组比较均有高度显著性差异（Ｐ＜０．０１）。结论：ｃ－ｍｙｃ抑癌基因ｐ５３表达与涎腺上皮细胞增殖速度加快和突变有关，ｐ５３阳性率升高，虽与细胞恶变关系密切，但仅为判断细胞分化程度的一个参考指标。     

涎腺恶性多形性腺瘤病理形态及免疫组织化学研究

本文对８３例涎腺恶性多形性腺瘤进行形态观察，其中２０例用ＣＥＡ、Ｓ－１００蛋白和二种角蛋白进行免疫组化标记。认为恶性多形性腺瘤大多数由良性多形性腺瘤恶变而来，瘤细胞浆内ＣＥＡ阳性是诊断恶性的依据之一。并提出将恶性多形性腺瘤分为癌和癌肉瘤二个组织学亚型。     

c－erbB－2的表达与涎腺恶性多形性腺瘤预后的关系

本研究的目的在于观察癌基因ｃ－ｅｒｂＢ－２表达与涎腺恶性多形性腺瘤患者预后的关系。采用抗ｃ－ｅｒｂＢ－２癌蛋白单克隆抗体ＡＢ－３，应用免疫组织化学方法，对６０例涎腺恶性多形性腺瘤中ｃ－ｅｒｂＢ－２癌基因表达产物进行了检测。结果发现，２５例ＭＰＡ显示ｃ－ｅｒｂＢ－２表达产物阳性，阳性表达率４１．６７％。结合临床随访资料分析，ＭＰＡ复发组ｃ－ｅｒｂＢ－２阳性表达率明显高于未复发组（Ｐ＜０．０５）；ｃ－ｅｒｂＢ－２表达阳性患者术后５年生存率明显低于表达阴性者（Ｐ＜０．０５）。表明ｃ－ｅｒｂＢ－２癌基因表达与ＭＰＡ患者的预后密切相关，检测ｃ－ｅｒｂＢ－２表达产物可能成为估价ＭＰＡ预后的可靠指标之一。     

涎腺肿瘤中c-erbB-2癌基因扩增及蛋白产物过度表达的研究

作者采用ＤＮＡ斑点杂交及免疫组织化学方法对２８５例涎腺肿瘤石腊包埋组织中ｃ－ｅｒｂＢ－２癌基因扩增及蛋白产物过度表达进行检测．结果表明：涎腺恶性肿瘤及多形性腺瘤有不同程度的ｃ－ｅｒｂＢ－２癌基因扩增,其中癌在多形性腺瘤中扩增率２０／３０（６６．６７％）,腺癌为１３／２０（６５．００％）,明显高于其它肿瘤（ｐ＜０．０５）．ｃ－ｅｒｂＢ－２蛋白产物过度表达出现在癌在多形性腺瘤中１６／３０（５３．３３％）、腺癌１１／２０（５５．００％）、肉瘤１／１０（１０．００％）及多形性腺瘤中５／３５（１４．２９％）,癌在多形性腺癌中及腺癌表达率明显高于其它肿瘤（ｐ＜０．０５）．提示：ｃ－ｅｒｂＢ－２癌基因可能与涎腺肿瘤细胞分化程度有关,在部分肿瘤的发生、发展过程中起一定的作用．     

非同位素PCR-SSCP-EB染色技术检测头颈部鳞癌p53基因点突变

应用非同位素ＰＣＲ－ＳＳＣＰ－ＥＢ染色技术检测新鲜头颈部鳞癌组织中ｐ５３基因点突变。结果显示１３例标本中７例有异常电泳带，其中位于第５外显子者１例，位于第６，７，８外显子者各２例，说明头颈部鳞癌的发生与ｐ５３基因突变有密切关系。与传统的同位素ＰＣＲ－ＳＳＣＰ相比，非同位素ＰＣＲ－ＳＳＣＰ－ＥＢ染色技术是一种更简便、安全、快速、有效的检测基因点突变的方法，适用于大量标本基因点突变的筛选性检测。     

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.     

Mutated and wild-type p53 expression and HPV integration in proliferative verrucous leukoplakia and oral squamous cell carcinoma

The frequencies of overexpression and mutation in the p53 tumor suppressor gene were examined in proliferative verrucous leukoplakia and oral squamous cell carcinoma with immunohistochemistry and single-strand conformation polymorphism analysis of DNA fragments amplified by polymerase chain reaction. Ten samples each of normal oral mucosa, proliferative verrucous leukoplakia, and squamous cell carcinoma were immunostained with antibodies against p53 protein; 8 of 10 cases of proliferative verrucous leukoplakia cases and 7 of 10 cases of oral squamous cell carcinoma were positive for p53 protein. Minimal staining was observed in normal oral tissues. The quantified labeling indexes demonstrated a range that corresponded to lesion progression. Single-strand conformation polymorphism analysis revealed p53 gene mutations within exons 5 to 8 in 40% (4 of 10) of the squamous cell carcinoma samples. Two of the 4 mutated squamous cell carcinoma samples lacked p53 expression. No p53 mutations were detected in proliferative verrucous leukoplakia tissues. Human papillomavirus 16 was identified in 2 of 7 p53 positive oral squamous cell carcinoma samples. Human papillomavirus 16 and 18 were identified in two of eight p53 positive proliferative verrucous leukoplakia samples. One p53 negative squamous cell carcinoma sample was positive for human papillomavirus 16 and had a mutation in exon 6 of the p53 gene. Human papillomavirus infection along with p53 expression plays a yet to be defined role in the pathogenesis of a limited number of cases of proliferative verrucous leukoplakia and squamous cell carcinoma. p53 Immunohistochemistry, p53 gene mutations, and human papillomavirus infection prevalence do not provide a means to differentiate between leukoplakia and carcinoma and do not provide a predictive test for progression of leukoplakia to carcinoma.     

p53 aberrations in oral submucous fibrosis and oral squamous cell carcinoma detected by immunocytochemistry and PCR-SSCP

Trivedy C, Warnakulasuriya KAAS, Tavassoli M, Steingrimsdottir H, Penhallow J, Maher R, Johnson NW: p53 aberrations in oral submucous fibrosis and oral squamous cell carcinoma detected by immunocytochemistry and PCR-SSCP. J Oral Pathol Med 1998; 27: 72–7. © Munksgaard, 1998.
An archival series of oral biopsies from Karachi, Pakistan, consisting of 21 cases of oral submucous fibrosis (OSF) and 27 cases of squamous cell carcinoma (SCC), of which 6 had arisen from OSF, were used to examine the aberrations in the structure and expression of the p53 tumour suppressor gene. The PCR-SSCP method was used for mutation analysis of exons 2–9, and (over)expression of p53 protein was detected by immunocytochemistry using monoclonal antibody DO 7. Positive immunostaining was observed in 15/20 (75%) of OSF specimens, 3/6 (50%) of SCC arising from OSF and 14/21 (67%) of SCC not arising from OSF. Mobility shifts in SSCP indicative of a mutation in p53 or loss of heterozygosity (deletion of a band) were seen in 13/21 cases of OSF and 15/27 cases of SCC. There was concordance between immunocytochemistry and SSCP results in a majority (33/48) of samples. Though the number of analysed SCC cases arising from OSF was limited, the results suggest that p53 mutation/protein stabilisation may play a part in the pathogenesis of OSF and its progression to SCC.     

Expression and mutations of p53 in salivary gland tumours

A series of 219 salivary gland tumours (103 carcinomas and 116 benign tumours) were analysed for p53 protein expression using immunohistochemistry, and for mutations in p53 gene using non-radioactive single strand conformation polymorphism (SSCP). p53 expression was present in 36% (42/116) of the benign tumours and in 54% (56/103) of the carcinomas. The highest prevalence of p53 expression was found in adenoid cystic carcinomas (69%). followed by mucoepidermoid carcinomas (67%). Of the benign tumours, pleomorphic adenomas showed the highest prevalence of p53 positivity (41%). In malignant tumours, expression of p53 bore no correlation to local recurrence, metastatic disease or survival of the patients. Exons 5 through 9 were analysed and four mutations were found in 20 cases of p53-immunopositive tumours and two in 20 p53-negative tumours. Each of the exons 5.6 and 8/9 had two mutations, whereas no mutations were detected in exon 7.     

一个掌跖角化-牙周破坏综合征患者家系组织蛋白酶C基因突变分析

目的 对一个掌跖角化-牙周破坏综合征（PLS）患者及其家系组织蛋白酶C基因（CTSC）突变位点进行分析,探讨PLS的分子致病机制。方法 提取1例PLS先证者及其直系血亲（父母、弟弟）的基因组DNA,应用聚合酶链反应和DNA直接测序技术分析先证者及其直系血亲CTSC基因的突变情况。结果 PLS先证者CTSC基因存在复合型杂合突变。先证者位于外显子6的第800位碱基发生了一个杂合错义突变,该碱基对中的碱基T被C取代（c.800T>C）,其编码的氨基酸由亮氨酸改变为脯氨酸（p.L267P）;位于外显子7的第1015位碱基发生了一个杂合错义突变,该碱基对中的碱基C被T取代（c.1015C>T）,其编码的氨基酸由精氨酸改变为半胱氨酸（p.R339C）。其中,c.800T>C来自母亲,c.1015C>T来自父亲,弟弟的CTSC基因未见突变。结论 PLS的临床表征与CTSC基因突变有关。     

Association of muscle segment homeobox gene 1 polymorphisms with nonsyndromic cleft lip with or without cleft palate

OBJECTIVE: Muscle segment homeobox gene (MSX)1 has been proposed as a gene in which mutations may contribute to nonsyndromic cleft lip with or without cleft palate (NSCL/P). To study MSX1 polymorphisms in NSCL/ P by means of polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP), and investigate the association of MSX1 exons 1 polymorphisms with NSCL/P. METHODS: DNA were extracted from blood samples from NSCL/P and unrelated normal subjects. Genome DNA from peripheral leukocyte with these blood samples were extracted, which was used as template to amplify desired gene fragment of MSX1 exons 1 by means of polymerase chain reaction (PCR). The PCR products were examined by single-strand conformation polymorphism (SSCP). The MSX1 exons 1 polymorphisms were examined by sequencing if mutations were found. RESULTS: MSX1 genes of exon 1 mutation was not been found in the NSCL/P and unrelated normal subjects by SSCP. CONCLUSION: No correlation between MSX1 exon 1 and NSCL/P was found. MSX1 exon 1 may not be a key gene (susceptibility gene) in NSCL/P.     

Infrequent p53 mutations in patients with areca quid chewing-associated oral squamous cell carcinomas in Taiwan

Mutations in the conserved regions (exons 5-9) of the p53 gene were investigated in 37 untreated human primary oral squamous cell carcinomas (SCCs) using polymerase chain reaction-single strand conformation polymorphism and DNA sequencing analyses. P53 mutations were detected in 2 of 37 (5.4%) oral SCC cases. One tumor sample (case 23) showed a mis-sense point mutation at codon 177, changing CCC to CTC, which resulted in a substitution of proline to leucine in the p53 protein. The other tumor (case 33) had a point mutation at codon 266, changing GGA to AGA and causing a substitution of glycine to arginine in the p53 protein. These two patients with p53 mutations did not have an areca quid chewing habit. These results suggest that mutations in the p53 gene may not play a role in the pathogenesis of human oral SCCs in Taiwan. Recently, we have shown that positive p53 staining was observed in 47 of 81 (58%) cases of oral SCC. The discrepancies between positive p53 protein staining and the low prevalence of p53 mutation in oral SCCs indicate that other mechanism(s) are involved in p53 overexpression.     

TP53 mutations in clinically normal mucosa adjacent to oral carcinomas

J Oral Pathol Med (2010) 39 : 662–666 Background: The tumour‐suppressor protein p53 often accumulates in histologically normal epithelium adjacent to oral squamous cell carcinomas (OSCC). We investigated whether this was associated with mutations in TP53, the gene for p53, and might implicate impending malignancy. Methods: Specimens from 18 human squamous cell carcinomas were stained with monoclonal p53 antibodies. Positive cells were microdissected with laser‐captured microscopy from the tumour and adjacent normal and dysplastic epithelium. DNA was extracted, and exons 5–9 of the TP53 gene were amplified by PCR. Amplified products were separated by denatured gradient gel electrophoresis. Fragments with a deviant DGEE pattern were sequenced. Results: TP53 mutations were found in six of 18 tumours. Fourteen specimens contained histologically normal mucosa adjacent to the tumour; 13 of these showed small clusters of p53 positive cells. Seven specimens contained both histological normal and dysplastic epithelial tissues adjacent to the tumour. A TP53 mutation was found in only one specimen; this mutation appeared in the normal mucosa, the adjacent tumour, and the epithelial dysplasia. Conclusion: We found that upregulation of p53 was a frequent event in histological normal mucosa adjacent to OSCC; however, it was rarely associated with a mutation in the TP53 gene.