应用基因芯片筛查维吾尔族妇女子宫颈癌相关基因

目的 探讨新疆维吾尔族妇女子宫颈鳞癌相关基因群的表达和功能。方法 应用含有2048条人类全长基因的cDNA表达谱基因芯片,对3例临床切除的宫颈癌及自身正常宫颈组织标本的基因表达谱进行分析。结果 在3例宫颈鳞癌组织中有共同差异表达的基因64条,其中表达量减少(下调)的基因11条,表达量增加(上调)的基因53条。结论 应用基因芯片技术可以快速、高通量的筛查出维吾尔族妇女宫颈鳞癌的相关基因。     

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。     

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.     

CIITA、LGMN、CTSB和GILT基因在子宫颈癌免疫逃逸中的作用

目的通过检测抗原呈递过程中CIITA、LGMN、CTSB、GILT基因表达模式的变化,探讨抗原呈递过程在宫颈癌免疫逃逸机制中的作用。方法收集子宫颈鳞癌、子宫颈上皮内瘤变(CIN)2和正常宫颈组织石蜡包埋标本各7例。采用QuantiGene2.0Reagent System定量检测CⅡTA、LGMN、CTSB和GILT基因的表达,cluster3.0聚类分析其基因表达模式特征。结果 CⅡTA、GILT基因在宫颈癌和CIN组织中呈较高表达,正常宫颈中呈较低表达;LGMN基因在宫颈癌和CIN组织中呈较低表达,而在正常宫颈中表达稍高;CTSB基因表达变异较大,在异常组织中表达可上调或下降,而正常组织中表达中等。结论四个基因可显示免疫系统抗原呈递的特征,可能区分CIN和宫颈鳞癌,有助宫颈癌的预诊,但由于例数少,有待进一步验证。     

应用cDNA表达谱芯片研究子宫内膜癌基因的表达谱

目的:应用cDNA表达谱芯片研究不同分化程度人子宫内膜癌组织的基因表达谱,分析子宫内膜癌发生、发展过程中基因表达的变化。方法:用一步法抽提5例高分化和4例低分化人子宫内膜癌组织和36例正常子宫内膜组织的总RNA,在反转录合成cDNA探针的同时以Cy3和Cy5标记肿瘤组和正常对照组,将Cy3和Cy5标记探针混合后与含有13 824个基因的人表达谱芯片杂交。经洗片、扫描、软件处理后,分析子宫内膜癌和正常子宫内膜对照之间基因表达谱的差异。对高分化和低分化子宫内膜癌基因表达谱进行聚类分析。结果:子宫内膜癌组织和正常对照基因表达谱比较,差异2倍以上基因共有26条,表达上调和下调的基因分别为2条和24条。聚类分析显示,通过基因表达谱基本可将高分化和低分化子宫内膜癌分开。结论:通过基因表达谱差异研究可以发现与子宫内膜癌发生、发展相关的基因;癌基因表达谱聚类分析可能会帮助判断高危和低危子宫内膜癌,有利于子宫内膜癌患者个体化治疗方案的制定。     

细胞质胸苷激酶在维吾尔族妇女宫颈癌的表达及临床意义

目的:探讨细胞质胸苷激酶(TK1)在宫颈鳞癌的表达及其临床意义。方法:制备40例维吾尔族妇女子宫颈癌及正常宫颈组织中总RNA,用RT-PCR方法分析TK1基因mRNA的表达水平,用免疫组化(SP)法检测其相应蛋白表达。结果:(1)维吾尔族妇女宫颈癌组织中TK1 mRNA及蛋白阳性检出率分别为67.5%及60.5%,对照组宫颈组织中检出率为26.7%及8.3%。宫颈癌组织中TK1 mRNA及蛋白阳性表达明显高于对照组的宫颈组织(P0.05);(2)在宫颈癌组织中TK1 mRNA及蛋白表达:不同临床分期及分化程度之间无显著差异(P0.05),但是,有淋巴结转移患者组织中阳性表达明显高于无淋巴结转移者,肿瘤直径≥4cm患者阳性表达明显高于肿瘤直径4cm的患者(P0.05)。结论:TK1在维吾尔族妇女宫颈鳞癌中高表达,对宫颈癌的诊断有重要的参考价值,TK1高表达可能与宫颈癌的进展及预后有关。     

CD24在早期宫颈癌中的作用机制及临床价值探索

目的:探究分化抗原簇蛋白24(CD24)在早期宫颈癌发生、发展中的作用及临床价值。方法:通过TCGA门户网站进行差异基因分析、基因筛选以及临床预后分析;通过免疫组织化学验证筛选基因在宫颈病变及早期宫颈癌患者间的表达差异;通过string数据库进行蛋白互作关系分析,通过cytoscape实现蛋白互作网络以及核心基因的筛选,并通过R语言实现GO富集分析以及KEGG富集分析。结果:根据TCGA差异分析发现CD24与早期宫颈癌发生、发展相关,其在早期宫颈癌组的表达水平显著高于正常组(P<0.05),而在早期宫颈癌组与阴道宫旁浸润组、早期宫颈癌组与远处转移组间差异无统计学意义(均P>0.05),该基因的表达水平可影响宫颈癌患者的总生存期(P<0.05)。CD24在早期宫颈癌患者的表达呈强阳性,而在高级别宫颈上皮内病变与正常宫颈组织分别为弱阳性及阴性表达。该基因表达与20个基因间存在蛋白互作网络关系,这些基因的功能主要富集于信号通路正调控及增殖相关功能,富集的通路与癌症通路以及磷脂酰肌醇3激酶-蛋白激酶B(PI3K-Akt)密切相关(均P<0.05)。结论:CD24对宫颈早期癌变过程具有重要作用,通过作用于表皮生长因子信号调控相关通路对早期宫颈癌细胞的增殖过程起到正向调控,其可能作为宫颈癌变的重要临床预测因子。     

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。     

hTERC基因表达在宫颈癌筛查中的意义

目的:探讨宫颈病变脱落细胞中人类染色体端粒酶基因(hTERC)的表达及hTERC基因检测在宫颈癌筛查中的临床意义。方法:收集宫颈病变患者的宫颈脱落细胞标本129例,依据组织病理学结果分为3组:宫颈上皮内瘤变低度病变组(CINⅠ)、高度病变组(CINⅡ/Ⅲ)、宫颈癌组(SCC);同时选择健康妇女的宫颈脱落细胞标本20例作为对照组建立实验室阈值;应用荧光原位杂交技术(FISH)检测两组标本hTERC基因的扩增情况,最终以病理学诊断为金标准,比较FISH检测结果。结果:研究各组中CINⅠ,CINⅡ/Ⅲ,SCC的hTERC基因阳性表达率分别为7.31%,56.25%,75.00%,hTERC基因扩增比例随病变进展而增加;研究组与对照组hTERC基因阳性表达率差异有统计学意义(P0.05);研究各组两两比较,hTERC基因阳性表达率低度病变组与高度病变组、宫颈癌组差异有统计学意义(P0.05),而高度病变组与宫颈癌组的差异无统计学意义(P0.05)。结论:FISH方法检测hTERC基因可用于筛查宫颈高度病变及宫颈癌,其阳性扩增信号多倍体型可能是宫颈病变进展的预测性指标。     

基因芯片在筛选卵巢癌基因中的应用研究

目的:研究人卵巢癌相关基因在卵巢癌组织和正常卵巢组织中的差异表达。方法:用含512条癌及抑癌基因的cDNA表达谱芯片研究一组卵巢癌组织样本的基因表达谱。结果:共筛选出差异表达与癌相关的基因54条,其中18条表达上调,36条表达下调。结论:用基因表达谱芯片研究基因谱,可有效的筛选出新的卵巢癌相关基因。     

Identification of chromosomal alterations important in the development of cervical intraepithelial neoplasia and invasive carcinoma using alignment of DNA microarray data

OBJECTIVE: To use microarray data to reveal regions of potential chromosomal loss or gain important in cervical intraepithelial neoplasia (CIN) and invasive cervical cancer by identifying mRNA expression biases in contiguous chromosomal regions. METHODS: Data from three RNA expression microarray experiments were used: one primary experiment using cDNA arrays profiling gene expression in cervical epithelium from viral cytopathic effect to invasive cancer, one experiment using Affymetrix arrays profiling gene expression in invasive cancerous cervical epithelium, and one experiment using Affymetrix arrays profiling gene expression in CIN cervical biopsy specimens. Gene expression was aligned along chromosomes to reveal regions of significant chromosomal imbalance. Regions showing significant gain or loss and verified in more than one experiment are presented here. RT-PCR was performed to validate expression of one gene in a region. RESULTS: Gain of 3q was detected from the CIN II (P=0.018), CIN III (P=0.005), and invasive cancer (P=0.0002) cDNA arrays, and gain of 12q was detected from the CIN (P=0.05) and invasive cancer (P=0.05) Affymetrix arrays. Loss of 6p was detected from the CIN III (P=0.004) cDNA arrays and invasive cancer (P=0.05) Affymetrix arrays. Loss of 4q was detected from the invasive cancer (P=0.04) cDNA arrays and invasive cancer (P=0.05) Affymetrix arrays. RAN, located in the region of gain on 12q24.3, was overexpressed in CIN and invasive cancer. CONCLUSIONS: Alignment of microarray expression data by chromosomes can be used to estimate regions of potential chromosomal aberration and identify differentially expressed genes important in the development of CIN and invasive cancer.     

不同期别子宫内膜癌组织中差异表达基因的层次聚类分析

目的探讨对不同期别子宫内膜癌组织中的差异表达基因进行层次聚类分析的意义。方法利用基因芯片技术分析32例不同期别子宫内膜癌组织中的差异表达基因,并对其基因表达谱进行层次聚类分析。结果根据国际妇产科联盟（FIGO）手术病理分期的不同对所有样本进行分组,分为Ⅰ、Ⅱ、Ⅲ、Ⅳ期组4组,筛选出与肿瘤转移相关的差异表达基因12个。根据这12个差异表达基因对32例内膜癌进行层次聚类分析,其结果与手术病理分期的符合率为66％。结论基因芯片技术可以发现与子宫内膜癌转移相关的差异表达基因,基因表达谱层次聚类分析可以在术前帮助判断高危型子宫内膜癌。     

Comparison of the molecular classification with FIGO stage and histological grade on endometrial cancer

PURPOSE OF INVESTIGATION: To classify endometrial cancers based on gene expression profiling, and to compare the prognostic value of the classification systems based on gene expression, grade, and stage. METHODS: cDNA microarray was carried out in 32 endometrioid endometrial cancers. Differentially expressed genes were identified among tumor tissues of different grades and stages. The classification and prognosis comparison analysis was performed between histological grades, FIGO stages and gene expression profiles. RESULTS: Class comparison analysis between different grade and stage endometrial cancer revealed 33 genes that are differentially expressed in tumors of different grades, ten in those of different stages, and 104 in a combined classification of grades and stages (p < 0.001). CONCLUSION: The cDNA microarray technique is a feasible way to generate gene expression profiles of endometrial cancer. Classification based on gene expression patterns may be more accurate than histological grade and FIGO stage classification in predicting the prognosis of tumors.     

Gene profiling in Pap-cell smears of high-risk human papillomavirus-positive squamous cervical carcinoma

OBJECTIVE: The purpose of the study was to investigate benign and malignant squamous cervical cells obtained by cervical swabs with regard to differentially expressed genes and gene expression profiling, in order to evaluate the biological behavior and clinical outcome of cervical malignancies. METHODS: Cervical squamous cells from six women with high-risk human papillomavirus positive [HR-HPV(+)] cervical carcinoma and from six HPV-negative women with normal ectocervical cells were analyzed by cDNA array. RESULTS: cDNA over-expression of several genes such as MET (c-met), Nm23-H1 (NME1), EGFR, KGFR, Nm23-H2 (NME2), ERBB2 (c-erbB-2), cyclin-dependent kinase inhibitor 4 (CDKN2A, p16INK4A), cytokeratin 8 (KRT8), KRAS (K-ras), FLT1, KGF (FGF7), BCL2-like 2 protein (BCL2L2), ERBB4, MYCN (N-myc), cyclin D1 (CCND1), KIT (c-kit), secreted phosphoprotein 1 (SPP1) and STAT1, was significant in cervical squamous cell carcinoma (CSCC). Gene expression was downregulated for 13 genes in CSCC, such as interleukin 1 alpha (IL1A), the transforming growth factor receptor beta superfamily (TGFbeta; TGFB), some members of the insulin-like growth factor binding proteins (IGFBPs) and the integrin family (ITGA6, ITGB1). CONCLUSION: This study was focused on the gene expression profiling of HR-HPV(-) and (+) cervical squamous cells and CSCC obtained by cytobrush. We observed gene expression patterns and signaling pathways that permit the investigator to distinguish between benign squamous cervical cells and CSCC with and without HPV infection.     

Differential gene expression profiles between tumor biopsies and short-term primary cultures of ovarian serous carcinomas: identification of novel molecular biomarkers for early diagnosis and therapy

OBJECTIVE: To identify novel molecular biomarkers useful for the early diagnosis and therapy of ovarian cancer by gene expression profiling. To compare the genetic fingerprints of flash-frozen ovarian serous carcinomas to those of matched highly purified primary tumor cell cultures. METHODS: Gene expression profiles of 19 flash-frozen ovarian serous papillary carcinoma (OSPC) were analyzed and compared to 15 controls (highly purified human ovarian surface epithelium short-term cultures, HOSE) using oligonucleotide microarrays complementary to >14,500 human genes. In addition, gene expression profiling of 5 highly purified primary OSPC cultured in vitro for less than 2 weeks was compared to flash-frozen ovarian carcinoma biopsies obtained from matched samples. Quantitative RT-PCR and IHC staining techniques were used to validate microarray data at RNA and protein levels for some of the differentially expressed genes. RESULTS: Unsupervised analysis of gene expression data readily distinguished normal tissue from flash-frozen OSPC and identified 901 and 557 genes that exhibited >3-fold up-regulation or down-regulation, respectively, in OSPC when compared to HOSE. Mammaglobin 2, an ovarian secreted protein, was identified as the top differentially expressed gene in OSPC (19 out 19 OSPC versus 0 out of 15 HOSE) with over 827-fold up-regulation relative to HOSE. The claudin and kallikrein family of proteins including the clostridium perfringens enterotoxin receptors claudin 3 and 4, kallikreins 6, 7, 8, 10, 11 and the immunomodulatory molecule B7-H4 were found among the most highly overexpressed genes in OSPC when compared to HOSE. Genetic fingerprints of flash-frozen OSPC were found to have high correlation with those of purified primary OSPC short-term in vitro cultures with only 31 out of 8,637 genes (0.35%) differentially expressed between the two groups. CONCLUSIONS: Short-term in vitro culture of primary ovarian carcinomas may greatly increase the purity of ovarian tumor RNA available for gene expression profiling without causing major alteration in OSPC fingerprints. Mammaglobin 2, kallikreins 6, 7, 8, 10, 11, claudin 3 and 4 and B7-H4 gene expression products represent candidate biomarkers endowed with great potential for early screening and therapy of OSPC patients.