前列腺癌与良性前列腺增生差异表达基因的研究

目的 :筛选和分析前列腺癌 (PCa)与良性前列腺增生 (BPH)组织中差异表达的基因。　方法 :应用含有 4 6 5个基因的寡聚核苷酸芯片 ,检测PCa和BPH组织中差异表达的基因。　结果 :PCa和BPH组织中表达差异有显著性的基因共 35个 ,PCa上调基因 1 7个 ,下调 1 8个。　结论 :PCa与BPH组织差异表达基因的研究 ,有助于了解PCa发生发展的分子机制和寻找PCa新的诊断治疗标志物。     

用基因芯片研究同一血管瘤增生和消退期差异表达基因

目的 利用基因表达谱芯片技术寻找血管瘤增生期到消退期之间差异表达的基因，初步探索血管瘤增生与自然消退的分子机制。方法 将4096条cDNA用点样仪点在特制玻片上制备成表达谱芯片；将同一血管瘤患儿的增生期和消退期瘤体组织的mRNA逆转录为cDNA、分别标记Cy3和Cy5两种荧光，制备成cDNA探针，与表达谱芯片杂交，通过计算机扫描、数据处理筛选出差异表达的基因。结果 差异表达的基因有194条，其中115基因条上调，79条下调：①增生期一些细胞因子和生长因子高表达；②消退期细胞凋亡因子表达增加；③血管形成和原癌基因参与血管瘤的增生；④线粒体激活的细胞凋亡通路和Writ／β-catenin通路可能与血管瘤的发生和发展有关。结论 血管瘤的发生可能是细胞增殖和凋亡比例失调所致。     

DMBT1基因在前列腺癌中表达的显著下调及临床意义

目的研究前列腺癌发生过程中可能受甲基化调节的基因,检测其在前列腺癌细胞系和组织中的表达并分析其表达水平与临床病理特征的关系。方法采用人类全基因表达谱芯片筛选出可能因去甲基化导致恢复表达的基因后,以实时荧光定量PCR检测其在前列腺癌细胞系LNCaP、PC-3及39例前列腺癌和16例前列腺正常组织中mRNA的表达情况。Western blot检测以上标本中目的蛋白的表达水平。结果在用甲基化抑制剂处理后,PC-3细胞系中DMBT1（deleted in malignant braintumor）基因表达显著上调,而在LNCaP细胞系中未见到显著改变。在100%的前列腺癌患者组织中DMBT1 mRNA及蛋白表达均显著下调。统计分析表明,DMBT1 mRNA的表达水平与cTNM分期及骨转移有关,而与年龄、PSA水平及肿瘤分化程度无显著关系。结论 DMBT1有可能成为指导前列腺癌分期及判断预后的分子标志物。但DMBT1的调控机制以及DMBT1在细胞生理和肿瘤发生中的作用还需要进一步的研究。     

基于基因芯片的前列腺癌转移高表达基因SPP1的生物信息学分析

目的:利用生物信息学方法分析前列腺癌转移高表达基因SPP1以及蛋白的结构,为进一步研究其功能和参与的调控机制提供一定的理论依据。方法:在公共基因芯片数据库(GEO)中下载前列腺癌转移相关基因芯片数据,利用BRB-Array Tools软件、protparam、Motif Scan、Signal P4.0、TMHMM、Net Phos2.0、Predict Protein、GO、KEGG、STRING等生物信息学工具进行数据挖掘及生物信息学分析。结果:共筛选出前列腺癌转移共同差异基因73个,表达上调21个,表达下调52个(P0.01),其中对前列腺癌转移高表达基因SPP1进行生物信息学分析发现,SPP1蛋白由314个氨基酸组成,该蛋白含有2个N-连接糖基化位点、8个酪蛋白激酶II磷酸化位点、3个PKC磷酸化位点,主要参与细胞外基质结合、骨化、成骨细胞分化、细胞黏附、PI3K-Akt信号通路、黏着斑、ECM受体相互作用、Toll样受体信号通路等分子功能和信号通路。结论:利用生物信息学的方法能有效分析基因芯片数据并获取生物内在信息,SPP1可能在前列腺癌转移中发挥重要作用,有望成为前列腺癌转移的早期诊断标志物和治疗的新靶点。     

前列腺癌组织中凋亡抑制基因Livin的表达研究

目的：探讨前列腺癌组织中凋亡抑制基因Livin的表达及其与临床病理特征的关系。方法：应用RT-PCR和免疫组化（SP）法，检测Livin在62例PCa组织中的表达情况，其中高分化癌14例，中分化癌30例，低分化癌18例，正常前列腺组织10例。结果：62例PCa组织中Livin基因高表达，正常的前列腺组织中Livin均无明显表达。62例PCa组织中Livin蛋白阳性表达共有37例（59．7％），低分化组Livin蛋白阳性表达率（83．3％）明显高于高分化组（28．6％），其差异有统计学意义（P〈0．05），中、高分化组间及低、中分化组间Livin蛋白阳性表达率无统计学差异（P〉0．05）。Livin蛋白阳性表达与PCa的临床分期比较，T1-T2 65．0％，T3-T4 77．3％，亦有统计学意义（P〈0．05），临床分期愈晚，Livin蛋白阳性表达率愈高。结论：Livin与PCa的发生、发展有关，检测PCa组织中Livin表达可能对判断PCa预后有一定意义。     

前列腺癌组织中凋亡抑制基因Livin的表达研究

目的:探讨前列腺癌组织中凋亡抑制基因Livin的表达及其与临床病理特征的关系。方法:应用RT-PCR和免疫组化(SP)法,检测Livin在62例PCa组织中的表达情况,其中高分化癌14例,中分化癌30例,低分化癌18例,正常前列腺组织10例。结果:62例PCa组织中Livin基因高表达,正常的前列腺组织中Livin均无明显表达。62例PCa组织中Livin蛋白阳性表达共有37例(59.7%),低分化组Livin蛋白阳性表达率(83.3%)明显高于高分化组(28.6%),其差异有统计学意义(P<0.05),中、高分化组间及低、中分化组间Livin蛋白阳性表达率无统计学差异(P>0.05)。Livin蛋白阳性表达与PCa的临床分期比较,T1~T2 65.0%,T3~T4 77.3%,亦有统计学意义(P<0.05),临床分期愈晚,Livin蛋白阳性表达率愈高。结论:Livin与PCa的发生、发展有关,检测PCa组织中Livin表达可能对判断PCa预后有一定意义。     

基于生物信息学途径探索前列腺癌的关键基因

目的:利用生物信息学方法探索影响前列腺癌发生的关键基因,以助于了解前列腺癌发生及发展的分子机制.方法:从GEO(Gene Expression Omnibus)数据库中下载微阵列数据集GSE70770、GSE32571及GSE46602,通过GEO2R在线分析工具筛选出正常前列腺组织与前列腺癌的差异表达基因(DEGs)...     

前列腺癌特异性基因的网络结构

目的探讨前列腺癌特异性基因的网络结构。方法将由基因芯片结果得到的所有769个上调基因和675个下调基因通过特异性通路技术分析,鉴别所有差异表达基因的网络结构。结果通过IPA软件分析,我们得到基因差异最大的3个网络。结论网络分析不但证实了蛋白的互相作用导致DNA复制、重组和修复的紊乱、细胞和细胞周期的损伤、遗传性疾病的发生和结缔组织损害,还观察到许多由Myc调控的基因参与了脂质代谢和核酸代谢的过程。     

前列腺癌患者血液中表达前列腺特异性抗原细胞的诊断意义

前列腺特异性抗原(PSA)测定已被广泛应用于前列腺癌的筛查。然而，血清PSA的低特异性可导致许多假阳性和假阴性结果以及临床上的不确定性。因此，迫切需要前列腺癌特异的诊断和预测指标。检测前列腺癌患血液和骨髓中表达前列腺特异性抗原的细胞(CPEC)在其分子诊断和预后中可能具有可行性。本研究采用RT-PCR-PSA法观察病变限于器官     

ELL基因在前列腺癌组织中的表达及其意义

目的分析 ELL基因在人类前列腺癌组织中的表达情况,探讨其在前列腺癌发生发展中的作用.方法收集45例前列腺癌组织、15例良性前列腺增生组织和15例正常前列腺组织,提取总 RNA,应用 qRT-PCR检测 ELL mRNA的表达情况,分析其与前列腺癌分级的关系.结果前列腺癌组织中 ELL mRNA 的表达量明显低于良性前列腺增生组织和正常前列腺组织(P＜0．05),而良性前列腺增生组织与正常前列腺组织间差异无统计学意义.随着前列腺癌Gleason评分的升高,ELL mRNA的表达呈下降趋势(P＜0．05).结论 ELL 基因在前列腺癌组织中呈低表达,其表达与前列腺癌的分级密切相关,提示其可能在前列腺癌的发生发展中发挥重要作用.     

Identification of differentially expressed genes associated with androgen-independent growth of prostate cancer

BACKGROUND: The human prostate cancer xenograft, CWR22, similar to most human prostate cancers, regresses after castration and recurs several months after the removal of androgen. Genes uniquely associated with proliferation were identified by comparison of tumors that exist in androgen absence but differ in proliferative capacity. METHODS: cDNA libraries from CWR22 tumors from 20-day castrate mice (proliferation undetectable) and recurrent CWR22 tumors (proliferation rate similar to androgen-dependent CWR22) were compared to evaluate the possibility that proliferation is triggered by either gain of function or loss of suppression. Differentially expressed genes were evaluated further for their temporal association with the onset of cellular proliferation using northern and western analysis and immunohistochemistry of a series of CWR22 tumors that spanned the transition from androgen-dependent to recurrent growth. RESULTS: Subtractive hybridization identified 11 candidate genes from among 1,057 clones examined. Northern analysis confirmed differential expression of 8 genes. Western analysis revealed an association between tomoregulin, translation elongation factor-1 alpha (EF-1 alpha), Mxi-1, and thioredoxin-binding protein 2/vitamin D up-regulated protein, and the onset of recurrent growth. Immunohistochemistry revealed expression of tomoregulin, EF-1 alpha, Mxi-1, and thioredoxin reductase-1 coincidental with the onset of cellular proliferation on day 120 after castration. CONCLUSIONS: One or more of these genes may represent an appropriate target to prevent, delay or treat recurrent prostate cancer.     

Identification of differentially expressed genes in organ-confined prostate cancer by gene expression array

BACKGROUND: To understand the molecular mechanisms underlying prostate cancer, we have utilized the gene expression array to search for genes whose expression is altered in this disease. METHODS: RNA quality from manual microdissected tissue was compared with that from microselected tissue by electrophoresis. For array analysis, malignant and normal prostate epithelium was enriched using microselection technique from prostate cancer and the peripheral zone of a normal prostate. Identical array membrane was hybridized to labeled cancer and normal cDNA, respectively. The differentially expressed gene was further evaluated by RT-PCR. RESULTS: Microdissection, but not microselection, causes visible degradation to RNA. Of the 588 genes on the membrane, 87 genes yielded significant signals. Based on a three fold difference relative to normal prostate tissue, 1 gene was overexpressed and 12 genes underexpressed in prostate cancer. Of them, five showed statistically significant reduction in mRNA levels in six prostate cancer specimens compared with seven normal prostate specimens. These five genes are glutathione S-transferase M1 (GSTM1), monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-alpha receptor-1 (TNFR-1), transforming growth factor beta3 (TGF-beta3), and inhibitor of DNA binding-1 (ID-1). CONCLUSIONS: GST-based metabolism, cytokine MCP-1 and TNFR-1, and TGF-beta3 signaling pathways, and some helix-loop-helix nuclear proteins could be potentially important in organ-confined prostate cancer and deserve further investigation.     

乳腺癌激素耐受细胞系LCC2的基因表达分析

目的　研究乳腺癌激素治疗耐受产生中相关基因群的表达及其功能。方法　用 80 0 0种人类基因PCR产物制成BioDoor80 0 0型表达谱芯片 ,分离纯化乳腺癌激素耐受细胞系LCC2和乳腺癌细胞系MCF 7mRNA ,制备表达谱探针 ,用ScanArray30 0 0荧光扫描仪扫描芯片荧光信号图像 ,利用计算机分析MCF 7细胞系和乳腺癌激素耐受细胞系LCC2之间差异表达的基因。结果　在 80 0 0种基因中 ,MCF 7细胞和乳腺癌激素耐受细胞系LCC2之间差异表达的基因有 1892条 (2 3 6 5 % )。生物信息学分析显示 ,这些差异表达基因可能与乳腺癌激素治疗耐受的产生有相关性。结论　对于乳腺癌相关基因群的研究有助于认识肿瘤发病机制和激素治疗耐受机制。     

Screening of differentially expressed genes and identification of AMACR as a prognostic marker in prostate cancer

Prostate cancer, the second most common cancer found in male over the world, was estimated to have 191,930 new cases and 33,330 deaths in 2020 in the United States. Prostate cancer is very common in male, about 12.1% of men will acquire this cancer in their lifetime, and a higher risk was reported in older men and African American men. Gene deregulations have been found to be extensively associated with cancer development. To gain further insight into how gene deregulation affects prostate cancer, we analysed three gene profiling datasets of prostate cancer from Gene Expression Omnibus (GEO) applying bioinformatic tools in our study. Firstly, we identified common differently expressed genes (DEGs) shared by the three gene profiling datasets, constructed protein–protein interaction network and determined top 10 hub genes. Further DEGs validation in TCGA and Human Protein Atlas Database identified AMACR as the core gene. We then analysed the role of AMACR in prostate cancer cell lines and found that AMACR-knockdown resulted in the decreased cell proliferation and increased apoptosis. These results suggest an oncogenic role of AMACR in prostate cancer, and it could be a potential biomarker for the diagnosis of prostate cancer.     

Interspecies comparison of prostate cancer gene‐expression profiles reveals genes associated with aggressive tumors

Prostate cancer (PC) is a heterogeneous disease whose aggressive phenotype is the second leading cause of cancer‐related death in men. The identification of key molecules and pathways that play a pivotal role in PC progression towards an aggressive form is crucial. A major effort towards this end has been taken by global analyses of gene expression profiles. However, the large body of data did not provide a definitive idea about the genes which are associated with the aggressive growth of PC. In order to identify such genes, we performed an interspecies comparison between several human data sets and high quality microarray data that we generated from the transgenic adenocarcinoma of mouse prostate (TRAMP) strain. The TRAMP PC mimics the histological and pathological appearance as well as the aggressive phenotype of human PC (huPC). Analysis of the microarray data, derived from microdissected TRAMP specimens removed at different stages of the disease yielded genetic signatures delineating the TRAMP PC development and progression. Comparison of the TRAMP data with a set of genes representing the core expression signature of huPC yielded a limited set genes. Some of these genes are known predictors of poor prognosis in huPC. Interestingly, the modulation of genes responsible for the invasive phenotype of huPC occurs in TRAMP already during the transition to prostate intraepithelial neoplasia (PIN) and onwards to localized tumors. We therefore suggest that critical oncogenic events leading to an aggressive phenotype of huPC can be studied in the PIN stage of TRAMP. Prostate 69:1034–1044, 2009. © 2009 Wiley‐Liss, Inc.     

基因芯片在筛选胆管癌相关基因差异性表达的应用研究

目的　应用cDNA芯片技术进行肝外胆管癌相关基因表达谱差异分析 ,筛选胆管癌相关基因。方法　按一步法分别抽提 6例胆管癌和正常人胆管黏膜总RNA、纯化 ,逆转录合成掺入荧光分子的cDNA链探针 ,与 10 68条人PCR微矩阵芯片杂交 ,扫描芯片荧光信号图像 ,计算机分析比较二种组织基因表达谱差异。结果　胆管癌与正常胆管黏膜的基因表达谱分析 ,发现有 194条基因表达差异 ,6组标本 47条与肿瘤相关的基因一致向上或向下表达 ,2 3条表达上调 ,2 4条表达下调。结论　基因芯片能快速筛选胆管癌相关基因 ,分析这些差异表达的基因能够阐明胆管癌复杂的生物学特性与基因表达之间的内在联系 ,并识别肿瘤的标记物。