胃粘膜肠化生组织中多种抑癌基因的杂合缺失

目的 探讨多种抑癌基因异常在胃粘怕化生组织中的作用。方法 应用ＰＣＲ－ＲＦＬＰ法检测了６０例肠化生组织中ＹＮＺ２２、ＡＰＣ、ＭＣＣ、ＤＣＣ基因杂合性失现象。结果 ＹＮＺ２２杂合性缺失率为１９．４％（６／３１例）,ＡＰＣ、ＭＣＣ、ＤＣＣ杂剑性缺失率分别为１２．０％（３／２５例）、１３．０％（３／２３例）、４．３％（２／４７）。将肠化生分为Ⅰ、Ⅱ型肠化生（Ｐ〉０．０５）。２例Ⅲ型肠化分别同时检出ＡＰＣ     

鼻咽癌染色体7q32区域微缺失的研究

目的 进一步明确鼻咽癌染色体７ｑ３２的Ｄ７Ｓ５００－Ｄ７Ｓ４９５附近区域高频率等位基因缺失的范围。方法 采用更高密度的微卫星标记位点,分析３０个鼻咽癌病例等位基因杂合性缺失的情况。结果 ３０例 患者中有１９例（６３．３％）存在杂合性缺失;Ｄ７Ｓ５００－Ｄ７Ｓ５０９－Ｄ７Ｓ４９５是高频率缺失集中的区域,共同缺失区在Ｄ７Ｓ５０９附近。结论 Ｄ７Ｓ５０９附近可能存在与鼻咽癌发病相关的抑癌基因。     

癌基因c-erbB2、c-myc和抑癌基因p16、p53在口腔鳞癌中的蛋白表达及其协同作用

目的：探讨癌基因c—erbB2、c-myc和抑癌基因p16、p53在口腔鳞癌(OSOC)中的蛋白表达及其协同作用。方法：用免疫组化结合图像分析对口腔鳞癌中4种基因的蛋白表达进行定性、定位、定量研究。结果：口腔鳞癌中c—erbB2、c—myc、p16和p53的蛋白表达阳性率依次为46．67％、60％、86．67％和63。33％。肿瘤部位不同,erbB2蛋白表达有显著性差异;腭癌和口底癌中的erbB2蛋白表达都明显高于唇癌和牙龈癌中的erbB2表达。c-myc蛋白表达与p16蛋白表达之间具有显著性相关。结论：以上4种基因的蛋白表达增高在口腔鳞癌发生发展中具有重要作用,c—erbB2蛋白过表达在腭癌和牙龈癌中具有更重要的意义;c—myc和p16蛋白表达间具有一定的内在联系。     

Wilson病患者及其杂合子亲属的脑电图研究

目的：探讨Wilson病及其杂合子的脑电图改变与临床的关系。方法：记录43例Wilson病患者及其60例杂合子亲属的脑电图。结果：43例Wilson病患者脑电图异常35例(81%)，以轻、中度异常改变为主，杂合子亲属中脑电图异常率为60%，子女组脑电图异常率及异常程度均高于同胞父母组，差异显著；杂合子亲属脑电图改变有年龄依赖性。Wilson病及杂合子脑电图改变均以弥漫性慢波化、阵发性慢活动和样放电等为主要表现。结论：脑电图可以作为判断肝损型及无症状型Willson病患者中枢神经系统临床下损害的有效指征；Wilson病杂合子脑电图改变与其年龄密切相关。     

Wilson病患者及其杂合子亲属的脑电图研究

目的:探讨Wilson病及其杂合子的脑电图改变与临床的关系。方法:记录43例Wilson患者及其60例杂合子亲属的脑电图。结果:43例Wilson病患者脑电图异常率达83.9％,以轻、中度异常改变为主,三组患者间异常率差异不明显(P>0.05);杂合子亲属中脑电图异常率达60.0％,子女组脑电图异常率及异常程度均高于同胞父母组,差异显著(P<0.05和0.01);杂合子亲属脑电图改变有年龄依赖性。Wilson病及杂合子脑电图改变均以弥漫性慢波化、阵发性慢活动和癫痫样放电等为主要表现。结论:1.脑电图可以作为判断肝损型及无症状型患者中枢神经系统临床下损害的有效指征;2.Wilson病杂合子脑电图改变与其年龄密切相关。     

食管鳞癌HPV感染与病毒致癌基因E6的关系

目的：探讨人乳头瘤病毒（ＨＰＶ）与食管鳞状细胞癌的关系。方法：采用多重引物多聚酶链反应（ＰＣＲ）的免疫组化技术、对１０４例食管鳞癌进行ＨＰＶ ＤＮＡ和病毒癌基因Ｅ６蛋白检测。结果：ＨＰＶ ＤＮＡ阳性者占５０．９６％（５３／１０４），其中ＨＰＶ１６型ＤＮＡ４９．０６％（２６／５３），ＨＰＶ１８型 ＤＮＡ５．６％（３／５３），ＨＰＶ６／１１ ＤＮＡ７．５％（４／５３）；两上或三个类型的混合感染占３７．     

一例Pah基因杂合缺失合并半合子突变病例分析

目的 对一例苯丙氨酸羟化酶(PAH)缺乏症患儿进行基因组测序,分析其Pah基因变异情况,以明确病因。方法 采用免疫荧光法对新生儿进行血苯丙氨酸(Phe)浓度检测,血Phe浓度>2.0mg/dL进行尿蝶呤谱分析以鉴别病因;进一步用基因测序技术对其静脉血进行Pah基因测序,测序数据与正常人基因组进行比对,分析基因变异情况。结果 该患儿血Phe浓度为46.88 mg/dL(2765.92μmol/L),正常新生儿Phe浓度<2.0mg/dL,苯丙氨酸/酪氨酸(Phe/Tyr)为49.65(参考范围0.1～1.5),尿蝶呤谱结果为52.08(正常新生儿：19.8～50.3);基因测序结果发现1个缺失突变和1个剪接突变,分别为：exon6杂合缺失和第6外显子c.611A>G剪接突变,其中exon6片段缺失在人类基因突变数据库(HGMD)中未报道;患儿根据治疗前最高血Phe进行分类为经典型PAH缺乏症,需低Phe饮食治疗,并定期监测血Phe浓度水平。结论 初步明确了该患儿的病因为杂合缺失合并半合子突变,Pah基因型为exon6杂合缺失/c.611A>G;并发现了1种未报道基...     

差别PCR及在癌相关基因扩增和缺失研究中的进展

基因的扩增与缺失是癌基因等活化、抑制基因等失活的常见机制之一，差别－ｄ－ＰＣＲ为上述遗传学改变了提供了简便并较敏感的非同位数技术，本文简述了近年严ｄ－ＰＣＲ方法学和用于肿瘤研究的进展。     

NOB1mRNA在人食管鳞癌组织的表达及其临床意义

目的 探讨NOB1 mRNA(nin one binding protein)在食管鳞癌(esophageal squamous cell carcinoma,ESCC)患者组织中的表达及其与临床病理参数之间的关系.方法 运用RT-PCR方法检测19例ESCC患者癌组织及对应的癌旁组织中NOB1 mRNA的表达情况,分...     

食管鳞癌预后因素的病理学和免疫组织化学研究

目的 探讨食管癌组织病理学特征和免疫蛋白表达的预后价值。方法 检测９７例原发性食管鳞癌手术切除标本,对肿瘤组织病理学特征和免疫蛋白表达与患者生存期的关系进行单因素Ｋａｐｌａｎ Ｍｅｉｅｒ分析和多因素Ｃｏｘ分析。结果 单因素Ｋａｐｌａｎ Ｍｅｉｅｒ分析显示,低分化肿瘤、浸润性生长、肿瘤侵及外膜、间质无淋巴细胞浸润、淋巴结转移、高ＴＮＭ分期、表皮生长因子受体（ＥＧＦＲ）阳性表达和ｎｍ２３阴性表达与患者     

Loss of heterozygosity of the APC gene in oral squamous cell carcinoma

The aim of this study was to investigate loss of heterozygosity (LOH) of the APC tumor suppressor gene loci, using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) in 40 cases of oral squamous cell carcinoma (OSCC). Observed informativity was 72.5% for APC exon 11 and 82.5% for APC exon 15. LOH at APC exon 11 was observed in 2 (6.9%) of 29 informative cases, and no LOH was observed for APC exon 15. Our results suggest that inactivation of the APC gene plays a minor role in the carcinogenesis of OSCC.     

肝细胞癌肿瘤抑制基因的杂合性缺失和微卫星不稳定性研究

目的　了解肿瘤抑制基因(TSG)杂合性缺失(LOH)与微卫星不稳定性(MSI)在肝细胞癌发生机制中的作用,并探讨其临床病理学意义。方法　采用显微组织切割基础上的DNA直接测序法,从92例手术切除肝细胞癌中筛选出36例信息性肝细胞癌进行6种TSG(APC、DCC、MCC、OGG1、p53和RB1)的LOH检测,对其中15例肝细胞癌进行13个多态性微卫星位点的LOH和MSI检测,并与临床病理学参数的相关性进行统计学分析。结果　TSG的LOH总发生率为41 .7% (15 /36),仅MCC基因未出现LOH。15例肝细胞癌中有9例(60% )发生微卫星LOH,占检测微卫星的46 .2% (6 /13),但无1例肝细胞癌出现MSI。若将APC、OGG1和DCC基因LOH作为Ⅰ型(n=7 ),将p53和RB1基因LOH作为Ⅱ型(n=8)进行统计学处理,则两组肝细胞癌的平均瘤体直径分别为( 2. 9 ±1 .7)cm和(7 .2 ±3 .4)cm (P<0 .01),两组患者术后平均生存期分别为( 72. 0 ±38 6 )个月和(51 .0±30. 4)个月(P<0 .05 )。肝细胞癌基因变异型与患者的年龄、性别、血清甲胎蛋白水平、HBsAg阳性率、合并肝炎/肝硬化、肝细胞癌分化程度和组织学类型之间无明显相关性。结论　在肝细胞癌发生的多阶段演进与多基因变异过程中,LOH路径所起的作用要比MSI路径更大。Ⅰ型基因变异(APC、OGG1和DCC)主要在肝细胞癌早期阶段起作     

Loss of heterozygosity of selected tumor suppressor genes in human testicular germ cell tumors

Human testicular germ cell tumors (TGCTs) are histologically heterogenous neoplasms with a variable malignant potential. Two main groups of germ cell tumors occur in men: seminomas and nonseminomas. In the present study, a set of four tumor suppressor genes was investigated in testicular cancers. CDH1, APC, p53, and nm23-H1 genes were tested for loss of heterozygosity (LOH). Thirty-eight testicular germ cell tumors (17 seminomas and 21 nonseminomas) were analyzed by PCR using restriction fragment length polymorphism or the dinucleotide/tetranucleotide repeat polymorphism method. An allelic loss of p53 at exon 4 was detected in five nonseminomas, whereas LOH of p53 at intron 6 occurred in one of the seminoma and two of the nonseminoma samples. Allelic losses of the APC gene were present in three seminomas and one nonseminoma, whereas one seminoma and three nonseminomas showed LOH of CDH1. The analysis of allelic losses showed no common structural genetic alterations in tumor tissues, although a different pattern of LOH was observed between the two main histological groups of TGCTs.     

利用比较基因组杂交技术分析食管癌组织中染色体基因组的变化特征

目的　探讨河南食管癌高发区居民食管癌发生发展的基因组变化特征。方法　应用比较基因组杂交技术分析52例原发性食管癌患者染色体基因组变化,按临床分期、有无淋巴结及远处转移进行分组比较。结果　在食管癌中3q、8q、5p、1q、6q、18p、20q的染色体基因组扩增和3p、1p、9q、19p、4p、8p染色体基因组丢失频繁( >20% )。3q、5p、1q、11q13 14的染色体基因组扩增和4pq、13q染色体基因组丢失与食管癌病理分期相关(P<0 .05)。8q扩增和4p丢失与淋巴结转移相关(P<0. 05)。2p扩增和4pq、l1q14 qter的丢失与远处器官转移相关(P<0 .05)。结论　染色体3q、8q、5p、1q、6q、18p和20q部位可能存在与食管癌变密切相关的癌基因, 3p、1p、9q、19p、4p和8p可能存在与食管癌变密切相关的抑癌基因; 3q、5p、1q、11q13 14扩增和4pq、13q丢失与食管癌的发展相关,而8q、2p扩增和4pq、11q14 qter的丢失是食管癌发展的晚期事件与食管癌转移相关,不同的基因参与了淋巴结转移和远处器官转移。     

早中期食管鳞状细胞癌中Notch1与血管内皮生长因子C的表达及其意义

目的 研究Notch1与血管内皮生长因子C(VEGF-C)在早中期食管鳞状细胞癌(ESCC)中的表达及其与临床病理特征的关系.方法 收集2007年5月至12月在本院行食管癌根治术的40例早中期ESCC和8例正常食管组织病理标本,免疫组化方法检测Notch1和VEGF-C的表达,并探讨其与临床病理特征的关系,分析Notch1和VEGF-C表达的相关性.结果 与正常食管组织比较,早中期ESCC组织Notch1表达阳性率下降[47.5%(19/40)比100.0%(8/8),P=0.006)],而VEGF-C表达升高[67.5%(27/40)比0.0%(0/8),P=0.000].高、中、低分化ESCC组织中Notch1表达阳性率分别为80.0%(8/10)、50.0%(7/14)、25.0%(4/16),Notch1表达阳性率随肿瘤分化程度降低而降低(P=0.023),但在肿瘤浸润程度和淋巴结转移中差异无统计学意义(均P＞0.05).高、中、低分化ESCC组织中VEGF-C表达阳性率分别为30.0%(3/10)、71.4%(10/14)、81.3%(13/16),VEGF-C表达阳性率随分化程度的降低而升高(P=0.024).淋巴结转移阳性组VEGF-C表达阳性率高于淋巴结转移阴性组[100.0%(16/16)比45.8%(11/24),P=0.000].与T2分期组比较,T3～T4分期组ESCC组织VEGF-C表达阳性率升高[82.1%(23/28)比33.3%(4/12),P=0.000].Notch1与VEGF-C表达呈负相关(r=-0.486,P=0.003).结论 Notch1在早中期ESCC中表现为抑癌基因,其异常低表达可能是引起VEGF-C异常高表达的因素之一.     

Identification of methylated-differentially expressed genes and pathways in esophageal squamous cell carcinoma

Methylation, as an epigenetic modification, can affect gene expression and play a role in the occurrence and development of cancer. This research is devoted to discover methylated-differentially expressed genes (MDEGs) in esophageal squamous cell carcinoma (ESCC) and explore special associated pathways. We downloaded GSE51287 methylation profiles and GSE26886 expression profiles from GEO DataSets, and performed a comprehensive bioinformatics analysis. Totally, 19 hypermethylated, lowly expressed genes (Hyper-LGs) were identified, and involved in regulation of cell proliferation, phosphorus metabolic process and protein kinase activity. Meanwhile, 17 hypomethylated, highly expressed genes (Hypo-HGs) were participated in collagen catabolic process, metallopeptidase and cytokine activity. Pathway analysis determined that Hyper-LGs were enriched in arachidonic acid metabolism pathway, while Hypo-HGs were primarily associated with the cytokine-cytokine receptor interaction pathway. IL 6, MMP3, MMP9, SPP1 were identified as hub genes based on the PPI network that combined 7 ranked methods included in cytoHubba, and verification was performed in human tissues. Our integrated analysis identified many novel genetic lesions in ESCC and provides a crucial molecular foundation to improve our understanding of ESCC. Hub genes, including IL 6, MMP3, MMP9 and SPP1, could be considered for use as aberrant methylation-based biomarkers to facilitate the accurate diagnosis and therapy of ESCC.     

凋亡诱导因子介导TAp63γ诱导的人食管鳞癌细胞株EC9706细胞凋亡

目的探讨TAp63γ诱导的人食管鳞癌细胞株EC9706细胞凋亡的分子机制。方法用免疫组织化学和逆转录聚合酶链反应（RT-PCR）检测食管鳞癌EC9706细胞中凋亡诱导因子（AIF）和p63的表达;将质粒pcDNA3．1-TAp63γ转入EC9706细胞,流式细胞仪检测凋亡率,并用激光共聚焦显微镜和亚细胞器分离技术观察分析AIF的转位;采用RNA干扰技术下调AIF的蛋白表达水平,caspase广谱抑制剂zVAD．fmk预处理细胞,再用流式细胞仪分析EC9706细胞表达TAp63γ后凋亡率的变化。结果EC9706细胞中AIF表达且定位在细胞质中,p63基因的表达亚型是ANp63,TAp63γ不表达;在pcDNA3．1-TAp63γ转染组和pcDNA3．1-p53转染组的细胞质及细胞核蛋白中分别检测到AIF,pcDNA3．1转染组的胞核蛋白无AIF信号;pcDNA3．1转染组、pcDNA3．1-TAp63γ转染组、AIF—siRNA干扰后再转染pcDNA3．1-TAp63γ组、zVAD．fmk处理后再转染pcDNA3．1-TAp63γ组和AIF-siRNA干扰联合zVAD．fmk处理后再转染pcDNA3．1-TAp631组的凋亡率分别为1．37％、13．64％、4．52％、4．03％和1．91％。结论TAp63γ在诱导EC9706细胞凋亡的过程中,AIF从线粒体释放入胞质并转位到细胞核内;下调AIF的蛋白表达水平可降低TAp631诱导的细胞凋亡;TAp631诱导的细胞凋亡依赖于AIF和caspase。     

Loss of heterozygosity of APC and CDH1 genes in laryngeal squamous cell carcinoma

The molecular mechanisms involved in the development and progression of laryngeal cancer, specifically squamous cell carcinoma, still need further investigation and elucidation. Twenty-two laryngeal squamous cell carcinomas were analyzed in our study regarding genetic changes of two tumor suppressor genes: Adenomatous polyposis coli (APC) and E-cadherin (CDH1). APC gene instability was tested by polymerase chain reaction (PCR)/loss of heterozygosity (LOH) using the restriction fragment length polymorphism (RFLP) method. The samples were also screened for mutations using the heteroduplex method. E-cadherin gene was analyzed by PCR amplification of tetranucleotide marker (D16S752) linked to E-cadherin gene. The results of our analysis showed three samples with LOH of the APC gene out of 15 heterozygous patients (20%). Only one LOH of the CDH1 gene (5.5%) out of 18 heterozygous patients was discovered. D16S752 marker did not reveal any replication error-positive samples. There were six samples showing heteroduplexes (33%) encompassed in APC's exon 11. Altogether, nine samples (41%) showed alterations of the APC gene. Our results suggest that alterations of APC gene may have a role in squamous cell carcinoma development. Detected LOH of the E-cadherin gene indicates that genetic changes of this gene are not very frequent, but that other components of the wnt signaling cascade may also be involved.     

Loss of heterozygosity at 6p21 underling HLA class I downregulation in Chinese primary esophageal squamous cell carcinomas

Loss or downregulation of human leukocyte antigen (HLA) class I molecules is a widespread mechanism used by tumor cells to avoid tumor recognition by cytotoxic T lymphocytes favoring tumor immune escape. Multiple molecular mechanisms are responsible for these altered HLA class I tumor phenotypes, such as the loss of heterozygosity (LOH) at chromosome region 6p21.3 . In this study, we used immunohistological techniques with a highly selective panel of anti-HLA monoclonal antibodies to analyze the expression of HLA class I molecules in 84 formalin-fixed, paraffin-embedded section and 49 frozen-fresh tissues of primary esophageal squamous cell carcinomas (pESCC) from Chinese patients. To elucidate the underlying mechanism of HLA class I loss or downregulation, we also analyzed LOH of previously selected microsatellite markers located in chromosomes 6 and 15 by polymerase chain reaction. DNA was obtained from frozen-fresh tumor tissues and surrounding stroma to define the LOH associated with chromosomes 6p21 and 15q21 . Our results showed that HLA-A, HLA-B/C, HLA class I heavy chain, β2-microglobuline, and HLA class I complex were lost or downregulated in pESCC ( P  < 0.0001), and were moderately associated with the microsatellite alterations in HLA class I gene regions, correlated with patients' age, tumor's location, and stage, and indicated that LOH at 6p21.3 is a frequent mechanism that leads to HLA class I abnormalities in pESCC.