食管腺癌与Barrett’s食管基因表达谱的研究

 目的 探讨从Barrett’s食管到食管腺癌的癌变过程中基因表达谱的变化。 方法 取同一食管腺癌患者大体手术标本经病理确诊的腺癌组织、Barrett’s 食管组织及正常组织,分别提取组织上皮的总RNA并纯化mRNA;将mRNA逆转录合成以Cy5和Cy3标记的cDNA链做探针,分别混合后在两张基因表达谱芯片上进行杂交。用扫描仪扫描芯片荧光信号,用软件对扫描图像进行分析。 结果 食管腺癌与正常食管上皮比较差异2倍以上共有214个基因;Barrett’s食管与正常食管上皮比较差异2倍以上共有90个基因。而Barrett’s食管组织和食管腺癌组织均下调的基因有24个,均上调的基因有21个。其中符合从Barrett’s食管到食管腺癌演变趋势的基因中,上调的有9个,下调的有18个。 结论 这些基因或其产物可作为Barrett’s食管具有癌变高危性的检测指标     

食管癌患者外周血基因表达谱研究

[目的]用基因芯片技术研究食管癌患者外周血和正常人基因表达谱差异。筛选与食管早期癌变相关的基因。[方法]分别抽提食管癌患者外周血细胞和正常人外周血细胞总RNA并纯化mRNA。将各mRNA逆转录合成以Cy5和Cy3标记的cDNA-链做探针，混合后在1张含有4096条双点人类全长基因的芯片上进行杂交，用扫描仪扫描芯片荧光信号图像，用软件对扫描图像进行数字化处理和分析。[结果]食管癌患者外周血细胞和正常人外周血比较差异2倍以上的共有92个基因，其中差异3倍以上共有36个基因，在92个基因中有4个基因与食管癌基因表达谱中出现的相同。[结论]利用基因芯片技术，进行了食管癌患者外周血基因表达谱的检测，发现有4个基因与食管癌基因表达谱中出现的相同，这些基因可能与人早期食管癌的发生有关。     

人卵巢浆液性乳头状囊腺癌相关基因表达谱

[目的]探讨人卵巢上皮性癌与正常卵巢组织基因表达谱差异。[方法]采用基因芯片技术，按一步法分别抽提10例卵巢浆液性乳头状囊腺癌患者的癌组织和正常卵巢组织的总RNA并纯化mRNA；逆转录合成以Cy5荧光标记卵巢癌细胞和Cy3荧光标记正常卵巢上皮细胞的cDNA作为探针，在含有8464点人类体细胞基因模板的表达谱cDNA芯片上进行杂交。用荧光扫描仪扫描芯片荧光信号图像，用计算机软件对扫描图像进行数字化处理分析。[结果]卵巢浆液性乳头状囊腺癌与正常卵巢上皮比较，差异5倍以上共有185个基因，其中表达上调（其荧光强度增强〉5）有86个，表达下调（其荧光强度减弱〈0.2）有99个。[结论]卵巢上皮性癌与正常卵巢上皮比较差异表达5倍以上的185个基因，可能与卵巢上皮癌的发生和发展有关。     

食管癌患者外周血基因表达谱研究(英文)

目的用基因芯片技术研究食管癌患者外周血和正常人基因表达谱差异,筛选与食管早期癌变相关的基因。方法分别抽提食管癌患者外周血细胞和正常人外周血细胞总 RNA 并纯化 mRNA;将各 mRNA 逆转录合成以 Cy5和 Cy3标记的cDNA一链做探针,混合后在1张含有4096条双点人类全长基因的芯片上进行杂交。用扫描仪扫描芯片荧光信号图像,用软件对扫描图象进行数字化处理和分析。结果食管癌患者外周血细胞和正常人外周血比较差异2倍以上的共有92个基因。其中差异3倍以上共有36个基因。有2个人尿激酶型纤溶酶原激活物受体(uPAR)基因显著上调,80K-L 蛋白基因、蛋白酪氨酸磷酸酶基因、原癌基因蛋白 mRNA 显著上调,胶原 V 型α-2基因显著下调。结论 80K-L 蛋白基因、蛋白酪氨酸磷酸酶基因、原癌基因蛋白、胶原 V 型α-2基因异常可能与食管癌发生、发展和转移相关。uPAR 基因可能被利用在外周血诊断食管癌微转移具有重要意义。     

食管癌患者外周血基因表达谱研究

目的用基因芯片技术研究食管癌患者外周血和正常人基因表达谱差异,筛选与食管早期癌变相关的基因。方法分别抽提食管癌患者外周血细胞和正常人外周血细胞总RNA并纯化mRNA;将各mRNA逆转录合成以Cy5和Cy3标记的cDNA一链做探针,混合后在1张含有4096条双点人类全长基因的芯片上进行杂交。用扫描仪扫描芯片荧光信号图像,用软件对扫描图象进行数字化处理和分析。结果食管癌患者外周血细胞和正常人外周血比较差异2倍以上的共有92个基因。其中差异3倍以上共有36个基因。有2个人尿激酶型纤溶酶原激活物受体(uPAR)基因显著上调,80K—L蛋白基因、蛋白酪氨酸磷酸酶基因、原癌基因蛋白mRNA显著上调,胶原V型α-2基因显著下调。结论80K-L蛋白基因、蛋白酪氨酸磷酸酶基因、原癌基因蛋白、胶原V型α-2基因异常可能与食管癌发生、发展和转移相关。uPAR基因可能被利用在外周血诊断食管癌微转移具有重要意义。     

细胞凋亡相关基因在食管上皮癌变中的表达及其意义

 目的　探讨食管上皮癌变过程中多个相关的细胞凋亡调控因子的表达状况及其意义。方法　应用碘化丙啶染色和间接免疫荧光标记方法,采用流式细胞术对60例食管癌组织及相应的癌旁组织进行定量检测。结果　bcl-2、c-FLIP基因蛋白在食管癌中的表达皆显著高于癌旁组织,两者比较差异有统计学意义（P＜0.01）;Fadd、Caspase-8和Caspase-3 蛋白在食管癌中的表达皆显著低于癌旁组织,两者比较差异有统计学意义（P＜0.01）。基因相关性比较结果显示,Fadd 与Caspase-8基因蛋白表达之间呈正相关关系（P＜0.01）,与正常黏膜组织和不典型增生组织相比,癌组织中DNA含量明显增高,异倍体细胞显著增加。结论　细胞凋亡的级联调控机制c-FLIP-Fadd-Caspase-8-Caspase-3-bcl-2在食管上皮癌变过程中起着重要作用。食管上皮癌变过程中,DNA含量及异倍体率增加。     

Cyclin E、CDK2和p21WAF1在食管上皮癌变过程中的表达及意义

目的探讨食管上皮癌变过程中细胞周期调控因子cyclin E、CDK2和p21WAF1的表达状况及其意义.方法应用免疫组化SP法和原位杂交方法分别检测48例食管癌组织、31例非典型增生组织和17例正常食管粘膜中cyclin E、CDK2和p21WAF1蛋白及mRNA表达.应用半定量RT-PCR和Western blot检测22例新鲜食管癌及相应癌旁组织的mRNA和蛋白表达.结果从食管正常粘膜、非典型增生组织到癌组织,cyclin E和CDK2蛋白和mRNA阳性表达率逐渐上升,差异具有统计学意义(P＜0.01或P＜0.05).食管癌组织中cyclin E、CDK2和p21WAF1蛋白及mRNA高表达,与癌旁组织或切缘正常食管粘膜有显著性差异(P＜0.01).cyclin E、CDK2和p21WAF1基因表达显著正相关(P＜0.01或P＜0.05).结论食管上皮癌变过程中,细胞周期相关基因cyclin E和CDK2表达逐渐增强.cyclin E基因表达异常是食管癌变过程中的早期事件.p21WAF1基因在食管癌中高表达,可能与细胞周期调控的反馈机制有关.     

高低转移人卵巢癌细胞系基因表达谱差异

目的 用基因芯片技术研究高低转移人卵巢癌细胞系（HO－8910PM和HO－8910）基因表达谱差异,筛选与转移相关的基因。方法 分别抽提高低转移人卵巢癌细胞和对照正常卵巢上皮的总RNA并纯化mRNA;分别将等量的mRNA逆转录合成以图像,用软件对扫描图像进行数字化处理和分析。结果 HO－8910细胞与正常卵巢上皮比较差异3倍以上共有355个基因;HO－8910PM细胞与正常卵巢上皮比较差异3倍以上共有323个基因。HO－8910PM与母系HO－8910比较差异2倍以上共有163个基因,差异3倍以上共有21个基因。结论 两株人卵巢癌细胞与正常卵巢上皮细胞基因表达谱存在差异,提示这些基因与卵巢癌的发生和发展有关;HO－8910PM与HO－8910比较存在差异的基因可能与高转移特性相关。     

人食管癌相关基因cDNA片段的克隆与初步鉴定

目的 分离人原发性食管组织中瓣的相关基因,揭示食管癌的易感性与癌变原理,方法 用高效,灵敏的ｍＲＮＡ差异显示技术,以２例正常食管上皮,１例癌旁上皮,２例高癌家族的食管癌组织互为对照,通过其对其因表达的比较,找出差异条带,进行ＲＴ－ＰＣＲ鉴定和ＤＮＡ序列分析,结果 （１）在实验中,分离,鉴定了１８个差异片段,其中包括正常组织不表达的突变食管基因（ｍｕｔａｔｅｄｅｓｏｐｈａｇｅａｌｇｅｎｅ,ＭＥＧ）５     

利用甲基化芯片及转录组测序方法筛选哈萨克族食管癌DNA异常甲基化谱

目的： 筛选新疆哈萨克族食管鳞癌DNA异常甲基化谱,为深入研究食管癌的发生机制提供线索。方法：采用Illumina Human Methylation 450K甲基化芯片,对6例哈萨克族食管癌组织及其癌旁正常组织进行全基因组甲基化检测,筛选DNA异常甲基化基因;利用Hiseq2000测序平台,对2例哈萨克族食管癌组织及其癌旁正常组织进行RNA水平检测,建立mRNA文库,并与DNA 甲基化结果进行关联分析,筛选DNA异常甲基化谱,同时对这些基因进行生物信息学分析。结果：食管癌组织中含高甲基化基因227个,低甲基化基因6个;癌组织mRNA表达量在0.031 2～8 192,癌旁正常组织在0.031 2～1 024。DNA异常甲基化基因与表达谱关联分析,呈负相关关系(即DNA高甲基化、mRNA低表达,或DNA低甲基化、mRNA高表达)的启动子区域基因共20个,包括启动子区域CpG岛高甲基化且表达下调10个位点、10个基因,低甲基化且表达上调11个位点、10个基因。生物信息学分析表明这些基因参与甲酰四氢叶酸分解代谢及调控细胞增殖等生物学过程,涉及一碳代谢通路及诱导细胞凋亡通路等。结论：初步建立了新疆哈萨克族食管癌DNA 异常甲基化谱,为哈萨克族食管癌的发病机制研究提供了新的思路。     

Study of the Gene Expression Profile of Human Ovarian Carcinoma by a Gene Chip

OBJECTIVE To study the difference in gene expression between human ovarian carcinoma and normal ovarian tissues, and screen the novel associated genes by cDNA microarrays.METHODS Total RNA from 10 cases of ovarian cancer and from normal ovarian tissues were extracted by a single step method. The cDNA was retro-transcribed from an equal quantity of mRNA derived from the 10 cases of ovarian carcinoma and normal ovarian tissues, followed by labeling the cDNA strands with Cy5 and Cy3 fluorescence as probes. The mixed probes were hybridized with BiostarH 8464 dot human somatic cell genes.Fluorescence signals were assessed by a ScanArray 4000 laser scanner and the images analyzed by Gene Pix Pro 3.0 software with a digital computer.RESULTS By applying the cDNA microarray we found a total of 185 genes for which expression levels differed more than 5 times comparing human ovarian carcinoma with normal ovarian epithelium. Among these genes 86 were up-regulated ＞5 times and 99 were down regulated ＜0.2.CONCLUSION The cDNA microarray technique is effective in screening the differential gene expression between human ovarian cancers and normal ovarian epithelium. It is suggested that these genes identified are related to the genesis and development of ovarian carcinoma.     

The gene expression profile of highly metastatic human ovarian cancer cell line by gene chip

In this paper, gene chip technique was used to analyze the difference of gene expression patterns in highly metastatic human ovarian tumor cell line HO-8910PM and in normal ovarian epithelial cells to explore the tumor-associated gene-cluster and its function in the process of occurrence an development of ovarian carcinoma. It will be helpful to comprehensively understand the molecular mechanism of cell transformation, to provide the molecular markers and target genes for clinical diagnosis a…     

Global analysis of altered gene expressions during the process of esophageal squamous cell carcinogenesis in the rat: a study combined with a laser microdissection and a cDNA microarray

The genetic alterations that occur during esophageal tumorigenesis have yet to be determined. We previously established a Wister rat carcinogenesis model of esophageal squamous cell carcinoma. To understand more about the molecular mechanisms during carcinogenesis, we produced esophageal neoplastic lesions by administering N-amyl-N-methylnitrosamine and 12-O-tetradecanoylphorbol-13-acetate to rats. We used laser microdissection to specifically isolate the cells from the normal epithelium, papilloma, dysplasia, and invasive carcinoma. Using a cDNA microarray representing 14,815 clones, we then analyzed the gene expression profiles for each esophageal lesion. The number of differentially expressed genes compared with the normal control dramatically increased in a step-by-step fashion from normal epithelium (1,151 +/- 119 genes) to papilloma (1,899 +/- 543 genes), dysplasia (1,991 +/- 193 genes), and invasive carcinoma (2,756 +/- 87 genes). A hierarchical clustering analysis showed that the three stages of normal epithelium, dysplasia (papilloma), and invasive carcinoma could be clearly classified, whereas the gene expression patterns of papilloma and dysplasia were indistinguishable. Using the Fisher criterion, we also identified 50 genes whose expression level had either significantly increased or decreased in a step-by-step manner from the normal epithelium to dysplasia and then finally to invasive carcinoma. Many of these genes were not previously known to be associated with esophageal carcinogenesis. The present findings in our rat model thus seem to provide us with a better understanding of the molecular alterations that occur during esophageal carcinogenesis and hopefully will also help lead to the development of novel diagnostic and therapeutic targets.     

Profiling of differentially expressed cancer-related genes in esophageal squamous cell carcinoma (ESCC) using human cancer cDNA arrays: overexpression of oncogene MET correlates with tumor differentiation in ESCC.

PURPOSE: To examine the global gene expression of cancer-related genes in esophageal squamous cell carcinoma (ESCC) through the use of Atlas Human Cancer Array membranes printed with 588 well-characterized human genes involved in cancer and tumor biology. EXPERIMENTAL DESIGN: Two human ESCC cell lines (HKESC-1 and HKESC-2) and one morphologically normal esophageal epithelium tissue specimen from the patient of which the HKESC-2 was derived were screened in parallel using cDNA expression arrays. The array results were additionally validated using semiquantitative PCR. The overexpression of oncogene MET was studied more extensively for its protein expression by immunohistochemistry in the two ESCC cell lines and their corresponding primary tissues and 61 primary ESCC resected specimens. Sixteen of these 61 ESCC cases also had available the corresponding morphologically normal esophageal epithelium tissues and were also analyzed for MET expression. The clinicopathological features associated with overexpression of the MET gene were also correlated. RESULTS: The results of cDNA arrays showed that 13 cancer-related genes were up-regulated > or =2-fold (CDC25B, cyclin D1, PCNA, MET, Jagged 2, Integrin alpha3, Integrin alpha6, Integrin beta4, Caveolin-2, Caveolin-1, MMP13, MMP14, and BIGH3) and 5 genes were down-regulated > or =2-fold (CK4, Bad, IGFBP2, CSPCP, and IL-1RA) in both ESCC cell lines at the mRNA level. Semiquantitative RT-PCR analysis of 9 of these differentially expressed genes, including the MET gene, gave results consistent with cDNA array findings. The immunostaining results of the expression of MET gene showed that MET was overexpressed in both ESCC cell lines and their corresponding primary tumors at the protein level, validating the cDNA arrays findings. The results of the clinical specimens showed that the MET gene was overexpressed in ESCC compared with normal esophageal epithelium in 56 of 61 cases (92%). Moreover, the overexpression of MET protein was more often seen in well/moderately differentiated than in poorly differentiated ESCC. CONCLUSIONS: Multiple cancer-related genes are differentially expressed in ESCC, the oncogene MET is overexpressed in ESCC compared with normal esophageal epithelium, and its protein overexpression correlates with tumor differentiation in ESCC.