卵巢癌相关基因筛选的初步研究

目的 通过研究人卵巢癌组织和正常卵巢组织中差异表达的基因，筛选出卵巢癌相关基因以用于早期诊断和治疗。方法 以cDNA基因表达谱芯片分析研究卵巢组织样本和正常卵巢组织的基因表达谱，计算机分析后比较两种组织中差异表达的基因。结果 共筛选出差异表达的基因1785条，表达上调基因有40条，下调基因138条，有显著表达差异的基因54条，涉及到11大类基因。结论 卵巢癌同其它恶性肿瘤一样，是多种基因结构和功能改变的结果，有关卵巢癌特异性相关基因及其作用尚有待进一步研究。     

雌孕激素受体基因及其相关基因在卵巢癌组织中的表达

目的应用基因芯片技术筛查卵巢癌雌孕激素受体基因及其相关基因表达。方法制备含有雌孕激素受体及其相关基因表达谱芯片，分别提取人正常卵巢组织及卵巢癌组织的mRNA，通过逆转录PCR方法，将^33P—dATP分别标记两种组织的cDNA纯化后与表达谱芯片进行杂交及扫描，用ImaGene3．0软件分析信号的强度和比值，筛选雌孕激素受体相关基因。对雌孕激素受体基因信号绝对值按早、晚期求均值及标准差筛选受体基因。结果筛选出4条雌孕激素受体基因、受体相关基因12条。结论通过基因芯片技术方法，初步筛出了卵巢癌雌孕激素受体基因及其相关基因。     

宫颈癌介入治疗前后敏感相关基因的表达谱研究

目的研究表达谱芯片筛选子宫颈癌介入治疗前后的差异表达基因,探究宫颈癌介入治疗的分子机制。方法取3例治疗有效的宫颈癌治疗前后的组织标本,采用基因表达谱芯片筛查差异表达基因,利用MAS软件挖掘差异表达基因的功能通路。结果介入治疗后和治疗前癌组织相比有209条基因表达上调,922条下降。功能分析示涉及细胞DNA修复、有丝分裂、BRCA1、BRCA2和ATR基因相关通路、乏氧相关因子等。结论表达谱芯片可用来快速高通量筛查可能与介入化疗栓塞敏感性相关的差异表达基因。     

应用基因芯片技术检测子宫肌瘤的基因表达谱

目的：研究人子宫肌瘤与子宫肌组织差异表达基因，寻找子宫肌瘤相关基因，以进一步阐明子宫肌瘤的发病机理。方法：以包含2048条cDNA基因表达谱芯片研究一组子宫肌瘤组织与子宫肌组织样本的基因表达谱。结果：在4例子宫肌瘤与子宫肌组织中三三比较共筛选出差异表达基因16条，主要涉及细胞信号和传递蛋白，蛋白翻译合成类基因。4组样本共同表达的差异基因有1条，属于蛋白翻译合成类基因。结论：基因表达谱芯片能有效地筛选子宫肌瘤相关基因，具有快速、高通量、高敏度等特点。     

应用基因表达谱芯片技术筛查中国汉族妇女宫颈癌相关基因的研究

目的 观察汉族妇女宫颈鳞癌(以下简称宫颈癌)基因表达谱的变化，研究宫颈癌发生、发展过程中相关基因，探讨基因表达谱技术在筛查宫颈癌相关基因中的应用价值。方法 应用含有2048条人类全长基因cDNA表达谱芯片，对3例临床手术切除的汉族妇女宫颈癌组织及自身部分正常宫颈组织标本基因表达谱进行分析。结果 3例汉族宫颈癌组织与正常宫颈组织差异表达基因分别为146、103、592条，主要涉及免疫相关基因、代谢基因、原癌基因、抑癌基因等，三组样本中共同差异表达基因2条，均为表达减少(下调)的基因为细胞信号和传递蛋白。结论 运用基因芯片分析基因表达谱，能快速、高通量、高敏度的筛选子宫颈鳞癌相关基因，并有效地对基因功能进行研究。     

应用基因芯片技术筛查卵巢癌相关基因

目的 探讨卵巢癌组织中相关基因的差异表达。方法 应用含有512条人类基因的cDNA芯片,检测卵巢癌组织和正常组织中相关差异表达的基因。结果 共筛查与卵巢癌相关的差异表达的基因37条,其中14条基因表达增加(上调),23条基因表达减少(下调)。结论 应用基因芯片技术,可筛查出新的卵巢癌相关基因。     

基因表达谱芯片在宫颈癌相关基因筛选中的应用研究

目的探讨基因表达谱芯片技术在高通量筛查肿瘤相关基因群及研究宫颈癌分子病理学变化中的应用价值.方法应用含有9 184条人类全长基因的cDNA表达谱芯片,对3例临床切除的宫颈癌及正常宫颈标本的基因表达谱进行分析.结果在3例宫颈癌和正常宫颈组织中均有差异表达的基因24条,其中在宫颈癌组织中上调基因12条,下调基因12条.结论运用本基因表达谱芯片对基因表达谱进行分析,能够有效筛查出新的宫颈癌相关基因.     

卵巢癌大网膜转移差异基因的筛选及验证

目的:通过筛选并验证卵巢癌原发灶和大网膜转移灶组织样本的差异表达基因,从中发现与卵巢癌大网膜转移相关的基因。方法:应用基因芯片检测3对配对的卵巢癌原发灶和大网膜转移灶组织样本差异基因表达谱,初步筛选出差异表达显著的基因,并通过qRT-PCR和免疫组化法验证基因芯片结果。结果:通过基因芯片检测共筛选得到187个差异性表达的mRNA,其中大网膜转移组相对于卵巢癌原发灶组表达上调的基因160个,表达下调的基因27个。初步筛选出差异表达显著的2个mRNA:Interleukin 8(IL-8)和Armadillo Repeat Containing 9(ARMC9);qRT-PCR和免疫组化结果显示,IL-8和ARMC9在大网膜转移灶中的表达均显著高于卵巢癌原发灶(P0.05)。结论:IL-8和ARMC9基因可能与卵巢癌大网膜转移有关。     

子宫肌瘤发病相关基因表达研究

目的:筛选子宫肌瘤发病相关基因。方法:分别提取10对子宫肌瘤及肌层组织mRNA,用Cy5-dCTP和Cy3-dCTP逆转录标记cDNA探针,然后与含14000条cDNA的表达谱芯片进行杂交,洗片。将所得芯片进行扫描、聚类分析,获得差异基因表达信息。结果:共得到39条表达差异基因,5条表达上调,34条表达下调。表达下调的基因主要与细胞周期、增殖与凋亡、DNA合成与转录及细胞信号转导相关。结论:子宫肌瘤的发生与发展可能与这些基因的表达失调相关。     

罗勒多糖对卵巢癌生物学行为的影响及其机制研究

目的:观察中药罗勒多糖对卵巢癌生物学行为的影响,探讨其抗卵巢癌作用的可能机制。方法:建立荷NuTu-19卵巢癌大鼠模型,观察罗勒多糖对荷瘤鼠肿瘤生物学行为的影响;抽提肿瘤组织总RNA,逆转录标记cDNA探针,用Cy3-dUTP标记对照组,Cy5-dUTP标记实验组,与表达谱芯片进行杂交,ImaGene3.0软件分析统计。同时取肿瘤组织,进行常规病理学检测。结果:与对照组相比,罗勒多糖治疗组肿瘤的生长及转移行为都受到明显的抑制:肿瘤数量减少,体积减小,腹水量显著减少(P<0.05);腹腔脏器转移程度积分显著降低(P<0.01)。罗勒多糖作用后,卵巢癌的基因表达谱发生了明显变化,应用表达谱芯片共筛选出168条差异表达基因。结论:罗勒多糖通过多个药效靶点抗肿瘤增殖及转移。     

Distinction between serous tumors of low malignant potential and serous carcinomas based on global mRNA expression profiling

OBJECTIVES: The molecular pathogenesis of ovarian serous tumors of low malignant potential (S-LMP) is not well understood, although the collective data suggest that they arise through molecular mechanisms distinct from those leading to conventional serous carcinomas (S-Ca). To further examine the molecular differences between these two diseases, we studied the gene expression pattern of ovarian S-LMP and S-Ca using high-density spotted cDNA and tissue microarrays. METHODS: Total RNA from 23 ovarian S-LMP and S-Ca was analyzed on 43,200 spot cDNA microarrays and the differential expression of proteins encoded by differentially expressed genes was validated using tissue microarrays. RESULTS: Unsupervised hierarchical clustering analysis of filtered data showed a complete separation between S-LMP and S-Ca, based predominantly on a small set of genes expressed at higher levels in S-LMP than in S-Ca. Many genes previously identified as up-regulated in ovarian carcinoma relative to normal ovarian tissue were expressed at even higher levels in S-LMP. These genes included mucin-1, mesothelin, HE4, PAX 8, and apolipoprotein J/clusterin. Immunohistochemical staining of tissue microarrays confirmed higher expression of selected proteins encoded by these genes in the S-LMP. Few genes were expressed at a higher level in S-Ca; these included E2F1, topoisomerase IIalpha, and cyclin E, with higher levels of cyclin E protein confirmed by immunohistochemistry. CONCLUSIONS: S-LMP and S-Ca are distinguished at the molecular level by a relatively small gene set, suggesting the pathogenesis of S-LMP as well as S-Ca may involve molecular pathways that escape detection by global gene expression profiling. In order to obtain biologically and clinically relevant information about the mechanisms involved in ovarian carcinogenesis, future studies based on molecular profiles of ovarian cancer should include analyses of low malignant potential tumors. Inclusion of such tumors is also critical to the evaluation of the efficacy of potential new diagnostic and/or therapeutic biomarkers.     

Characterization of differentially expressed genes in ovarian cancer by cDNA microarrays

The molecular events leading to the development and progression of ovarian carcinoma are not completely understood. We performed a large-scale survey for the identification of differentially expressed genes between ovarian carcinoma and normal ovarian tissue by using cDNA microarray analysis. We utilized 512 member human novel putative oncogene and tumor suppressor gene cDNA microarrays to study the differences in gene expression between ovarian carcinoma and normal ovarian tissues. Some differentially expressed genes have been further confirmed by immunohistochemical analysis. A total of 39 differentially expressed genes were identified, of which 16 and 23 were specifically expressed in ovarian cancer and normal ovarian tissue, respectively. The comparison of average signal of differentially expressed genes exhibited at least a twofold difference in expression. The differentially expressed genes may be related to the carcinogenesis and progression of the malignant growth. The use of cDNA microarrays allows simultaneous monitor of the expression of many genes, thereby it speeds up the identification of differentially expressed genes. It is essential for further exploration of the mechanisms of the disease.     

具有不同转移潜力的卵巢癌细胞系基因表达谱的比较研究

目的:比较具有不同转移潜力的上皮性卵巢癌细胞系基因表达谱,探讨卵巢癌侵袭转移过程中的关键基因和基因群,为最终揭示卵巢癌侵袭转移过程的分子机制奠定基础。方法:采用人工基底膜侵袭实验比较SKOV3·ip1细胞与其母系SKOV3细胞的侵袭转移能力;用17000点人类基因组cDNA芯片杂交检测两细胞系基因表达谱的差异;用RT-PCR方法验证基因芯片结果。结果:与其母系SKOV3相比,SKOV3·ip1细胞侵袭转移能力更强;基因芯片检测到1557个2倍差异表达基因,其中包括nm23-H2,c-erbB-2等重要的侵袭转移基因及一些差异显著的未知基因和EST片段;nm23-H2基因RT-PCR结果验证了基因芯片实验结果。结论:上皮性卵巢癌细胞系SKOV3.ip1具有更强的侵袭转移能力;卵巢癌的侵袭转移是一个涉及nm23,c-erbB-2等重要基因和其他未知新基因等多个基因共同参与的复杂过程;基因芯片是高通量基因筛选的有利手段。     

Expression profile of ovarian tumors: distinct signature of Sertoli–Leydig cell tumor

Epithelial ovarian tumors are the most common subtype of ovarian cancer. In this study, we reveal distinct expression signatures of previously uncharacterized ovarian carcinoma subtypes, including endometrioid component of mixed ovarian tumor and Sertoli-Leydig tumor. Both subtypes were compared to the most common and well-characterized ovarian epithelial carcinoma of the serous type. These comparisons were performed by complementaryDNA (cDNA) microarrays allowing high-fidelity measurements of the expression levels of 39,360 human individual cDNA species representing both known and unknown human genes. Functional analysis of differentially expressed genes in Sertoli-Leydig tumor revealed an upregulation in sonic hedgehog pathway, deregulation of several metabolic pathways especially in amino acid metabolism and overexpression of genes associated with protein synthesis, including ribosomal genes.     

基因芯片筛选上调miRNA-205表达后HeLa细胞相关基因

目的：应用基因表达谱芯片技术筛查上调微小RNA-205（miR-205）表达后人宫颈腺癌细胞株HeLa细胞中差异表达基因,并探讨其功能及上调miR-205致癌机制。方法：应用含有23 232条人类全长基因的cDNA表达谱芯片,对上调miR-205表达的HeLa细胞及HeLa细胞原株的基因表达谱进行分析。结果：与HeLa细胞原株相比,上调miR-205表达的HeLa细胞中有差异表达的基因共172条,其中34条上调,138条下调。结论：基因芯片技术能够有效筛查出上调miR-205表达后HeLa细胞中差异表达基因,miR-205引发宫颈癌是多因素多基因共同作用的结果。     

应用抑制性消减杂交筛选卵巢癌紫杉醇耐药差异表达基因

目的:应用抑制性消减杂交技术研究人卵巢癌紫杉醇耐药细胞株OC3/Tax300与其亲本细胞OC3(敏感株)之间基因表达的差异,筛选耐药相关基因,探讨基因表达差异与卵巢癌耐药的相关性。方法:以卵巢癌紫杉醇耐药细胞株OC3/Tax300 mRNA为检测子(tester),以其亲本细胞OC3(敏感株)mRNA为驱赶子(driver),构建cDNA消减文库。随机挑取文库克隆测序,所得结果在GenBank中做同源性比较分析。结果:成功构建了卵巢癌紫杉醇耐药细胞株的特异性cDNA消减文库。从文库中选取阳性克隆,其中76个测序成功,再随机选取36个序列与GenBank数据库进行初步比对,8个可能为新基因,1个为蛋白序列,另27个来源于16个已知基因,这些差异表达基因主要涉及细胞代谢、信号转导、细胞骨架、凋亡、蛋白翻译合成、发育等相关基因。结论:部分基因表达差异与卵巢癌紫杉醇耐药有关。