基因芯片筛选小鼠肝癌淋巴道转移相关基因

背景与目的肿瘤转移是一个多基因参与的复杂过程。由肿瘤转移所导致的恶性肿瘤患者死亡率高、预后差和其发生机制不清,长期以来一直是肿瘤学领域的突出问题。本研究利用基因芯片技术比较高低淋巴道转移能力小鼠肝癌细胞系Hca-F(高转移)和Hca-P(低转移)的基因表达谱,并筛选出与肿瘤淋巴道转移相关的基因。方法分别提取Hca-F和Hca-P细胞的总RNA,反转录合成双链cDNA,通过体外反转录合成生物素标记的cRNA探针,cRNA探针经片段化处理后分别与AffymetrixGeneChip誖MOE430A(包括22690个转录本对应于约14500个小鼠已知基因和4371个EST)杂交,杂交信号经扫描等处理并用生物信息学对检测结果进行分析。结果与Hca-P细胞相比较,Hca-F细胞中有901个(6.2%)基因表达上调幅度≥2倍,有129个(3%)EST上调幅度≥2倍。在公布的差异最显著的33个包括endoglin(EDG;CD105)、Mcam(Muc18;Mel-CAM;CD146)、Cdc42ep5(CEP5;Borg3)、Ptprr(proteintyrosinephosphatase,receptortype,R)、F2r[coagulationfactorⅡ(thrombin)receptor;Par1;ThrR]、D7Ertd458e(necl-5)、NR1D1、Serpinh1(HSP47)、AXL、Mak和Areg(AR)等基因表达中,上调幅度在29.86～13.93倍之间。根据GO(GeneOntology)分类和Treeview分析,这33个基因的功能主要为促血管生成、细胞粘附、信号转导、细胞运动、转录、分子伴侣活性、蛋白激酶活性和受体结合等。结论高通量的基因芯片技术筛选出大量淋巴道转移相关基因,对这些转移相关基因功能的验证有助于找到淋巴道转移的关键(代表)基因/通路,它们可作为肿瘤淋巴道转移诊断的指标和治疗的靶点。     

用基因芯片筛选胃黏膜癌变相关基因

Objective： To study the difference of gene expression and screen the carcinogenesis associated gene in gastric mucosa by oligonucleotide microarray. Methods： Using the U133A gene chip to detect the gene expression profile difference between pericancerous mucosa （mucosa inside nearly 2 cm by cancer） and normal section of gastric mucosa. Bioinformatics was used to analyze the detected result on their localization and function in chromosome. Results＂ （1） A total of 150 genes with a difference of more than 3 times in expression levels by comparing the pericancerous mucosa with normal gastric mucosa, of 130 genes were up-regulated （SLR〉I.5）, and 20 were down- regulated （SLR〈 -1.5）. From the gene expression difference was to do the function classification, among those 22 enzyme and 6 enzyme regular genes were most one （18.7%）. The next were 17 nucleic acid binding associated genes （11.3%）. The third were 15 signal transduction associated genes （10%）. Fourth, were 13 protein binding associated genes （8.7%）. Besides the 40 genes were unknown their function, above mentioned 4 groups were 48.7% of the gene total number; （2） The pericancerous mucosa （P） and gastric cancer （T） were simultaneously compared with normal gastric mucosa, which had 71 genes with the same expression difference, of 61 genes were up-regulated （pericancerous SLR〉I.5）, and other 10 genes were down-regulated （pericancerous SLR〈 -1.5）. From their localization on the chromosome, there was simultaneously 71 genes appearance both in the pericancerous mucosa and in gastric cancer. The most one was 11 abnormal genes on the No. 19 chromosome. The next was No. 1, 2, 16 and 17 chromosomes which had 6 genes, respectively. It was not finding an abnormal gene on the No. 5, 14, 22 and Y chromosome. Conclusion： It suggested those genes may be related to the promotion in early gastric carcinogenesis and their progress. Four main groups （enzyme and enzyme regular, nucleic acid binding, signal transduction, protein binding） that associated gene＇s abnormality be played an importance role in studying the carcinogenesis of gastric cancer. The No. 19 and No. 1, 2, 16, 17 chromosomes are important sites of the oncogene transformation.     

应用微矩阵表达谱基因芯片筛选食管癌新的相关基因

[目的]应用微矩阵表达谱基因芯片筛选食管癌新的相关基因.[方法]微矩阵表达谱基因芯片(14114种基因)筛选3例食管癌及其对照癌旁组织的差异表达基因,用Northern印迹证实,用末端终止法测序,将测序结果在GenBank数据库进行同源性检索.[结果]检测的3例临床标本中,共有的差异表达基因31条,上调基因10条,下调基因21条,其序列与Genbank数据库比较,从中筛选出2条无明显同源的人类已知基因或片段,即新基因,并列出其序列.[结论]微矩阵表达谱基因芯片可应用于筛选食管癌新的相关基因,其具备高通量、特异性好、快速的特点;食管癌和对照癌旁组织间存在2条新的差异表达基因.     

应用基因芯片技术筛选卵巢癌紫杉醇耐药差异表达基因

目的应用基因芯片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)之间的基因表达谱差异,筛选耐药相关基因,探讨基因表达谱差异与卵巢癌肿瘤耐药之间相关性。方法分别提取C3/Tax300与OC3细胞的总RNA并纯化mRNA;将等量的mRNA逆转录,以Cy5和Cy3标记的cDNA做探针,在Biostar H140S基因表达谱芯片上进行杂交。扫描芯片荧光信号图像,用基因图像分析软件对扫描图像进行数字化处理和分析。结果共筛选出显著表达差异基因134条,其中117种基因表达水平下调,17种基因表达上调,主要涉及细胞信号蛋白和传递蛋白,蛋白翻译合成类以及细胞骨架和运动、离子通道和运输蛋白、代谢、发育等14大类相关基因。下调基因主要为EBNA-3、COP9、StIP1,上调的基因主要是JAK2、HSPS、NADH等。结论这些基因表达差异与卵巢癌化疗耐药有关。     

基因芯片在早期预测肝癌肝移植转移复发中的应用

目的:通过基因芯片技术研究肝癌肝移植后移植肝发生转移的基因表达差异,寻找与转移相关的基因及其分子机制.方法: 选取30例因原发性肝癌行肝移植手术患者分为两组,对照组为肝移植手术后两年内未出现移植肝转移者,实验组为手术后两年内出现移植肝转移者.手术后3-24个月内在B超引导下行移植肝穿刺取材检测.结果: 两组差异表达的基因有96个,占芯片基因总数的2.34%,表达上调基因57个,表达下调基因39个.其中显著差异表达的基因21个,占差异表达基因的21.88%,上调基因13个,下调基因8个. 结论: 肝癌肝移植后移植肝发生转移是多种基因联合作用的结果.     

基因芯片筛选宫颈息肉炎性相关基因的研究

目的:运用基因芯片技术研究宫颈息肉组织炎症相关基因的表达谱,并探讨相关基因在宫颈息肉发病机制中的作用。方法:将按微距阵排列的509条炎症相关全长基因制成表达谱芯片。分别抽提8例宫颈息肉组织和宫颈黏膜组织的总RNA,逆转录成互补DNA(complementary DNA,cDNA)一链,并用cy5和cy3标记制备成杂交探针,混合后在3张炎症相关基因芯片上进行杂交,扫描仪扫描芯片荧光信号图像,对扫描图像进行数字化处理和分析。结果:宫颈息肉组织炎症基因表达谱中差异表达的基因共65条,其中上调基因35条,下调基因30条。3张芯片共存的差异基因16条,其中上调基因6条,下调基因10条。在差异表达的基因中,主要包括细胞因子及其受体、趋化因子及其受体、黏附分子、白细胞分化抗原、炎症信号传导分子,还包括一些补体分子及其受体。结论:宫颈息肉炎症相关基因表达谱中差异表达基因为研究宫颈息肉发病机理提供了线索,炎症反应和免疫应答可能是宫颈息肉形成和发展的重要机制。     

基因芯片在肾癌相关基因研究中的应用

目的：通过研究人肾癌组织和癌旁正常组织中差异表达的基因，寻找肾癌相关基因以用于诊断和治疗。方法：以包含8000个cDNA基因表达谱芯片研究1组肾癌组织样本的基因表达谱。按一步法抽提肾癌和对照组癌旁正常组织的总RNA并纯化mRNA；将等量的对照组织和肾癌组织mRNA分别逆转录合成荧光分子掺入的cDNA-链做探针，混合后杂交上述基因芯片。经严格洗片后用ScanArray3000扫描仪扫描芯片荧光信号图像，计算分析后比较2种组织中差异表达的基因。结果：共筛选出差异表达基因95条，其中未知基因44条。结论：基因芯片在筛选肾癌相关基因的改变具有快速、高通量、灵敏的特点，肾癌在细胞代谢、信号转录、蛋白合成的相关基因方面有异常表达。     

基因芯片筛选软骨肉瘤去分化相关基因初步探讨

目的：探讨用基因芯片技术区分普通型软骨肉瘤与去分化软骨肉瘤组织中基因的差异表达。方法：对病理证实的普通型软骨肉瘤及去分化软骨肉瘤组织,提取细胞mRNA,用寡核苷酸芯片,与正常关节软骨组织杂交后获得荧光信号,计算机软件分析荧光信号结果,并对数据进行归一化处理,分析组间显著变化的基因,然后对其进行聚类和主成份分析,筛选出差异表达的基因。结果：在所检测的人普通型软骨肉瘤与去分化软骨肉瘤组织中,31个基因有表达异常,其中高表达14条,低表达17条。结论：去分化软骨肉瘤与普通型软骨肉瘤间基因表达差异是明显的,这些差异基因主要分布于涉及TGT信号途径、Wnt信号途径、IHH/PThRP轴以及凋亡机制等多方面,且有些基因的作用目前尚不清楚。     

利用基因芯片技术筛选人卵巢癌耐药差异表达基因

摘 要：［目的］ 比较卵巢上皮癌铂类耐药和非铂类耐药细胞间的基因表达差异,并进行相关基因的筛选和鉴定。［方法］ 采用人类全基因组表达谱芯片对卵巢癌铂类耐药细胞SKOV3/CB和亲本细胞SKOV3进行差异基因的筛选和分析;用半定量逆转录聚合酶联反应(RT-PCR)对部分芯片结果进行核对验证。［结果］ 共分离筛选出差异基因3506个,SKOV3/CB比SKOV3上调的差异基因共有1712个,其中上调10倍以上的共有163个,下调的差异基因共有1794个,其中下调10倍以上的共有70个。SKOV3/CB比SKOV3上调10倍以上的3个差异基因,即ZNF198、ERCC5和BIRC3经半定量RT-PCR验证,表明上述3个基因在耐药细胞株中的表达的确高于敏感细胞株。［结论］ 卵巢癌的多药耐药是一个多种基因参与的复杂过程,ZNF198、ERCC5和BIRC3参与卵巢癌铂类耐药。     

基因芯片技术筛选Gemcitabine诱导胰腺癌Panc-1细胞凋亡相关基因的研究

目的研究抗癌药吉斯他滨（Gemcitabine）诱导胰腺癌Panc-1细胞凋亡的相关基因。方法应用基因芯片技术检测50．33μg／mL Gemcitabine作用于胰腺癌Panc-1细胞48h后凋亡基因表达的变化。结果Gemcitabine抑制细胞表达14种凋亡相关基因，包括IRF-4、Scotin、14．3．3gamma、PDE4、GRPS、TID1、AFP、STAT5、Sox-4、c-Rel、MYCN、Htt、CatD、Gd。相反，gemcitabine促进了3种凋亡相关基因的表达，包括CyclinB1、ASK1、AKT2。结论Gemcitabine诱导胰腺癌Panc-1细胞凋亡通过非依赖caspase ASK1途径。     

应用基因表达谱芯片从抗脱落凋亡肺癌A549细胞中筛选转移相关基因

背景与目的正常上皮或内皮细胞脱离细胞外基质会发生脱落凋亡,但肿瘤细胞可不依赖细胞基质生长而不发生凋亡,这一现象被称为抗脱落凋亡,目前国内外相关研究均认为抗脱落凋亡是肿瘤发生转移的始动环节。本实验旨在比较抗脱落凋亡及正常贴壁生长肺腺癌A549细胞基因组表达差异,从中筛选肺癌转移相关基因。方法利用多聚羟乙基甲基丙烯酸树脂处理培养皿,致细胞无法贴壁生长,建立抗脱落凋亡肺癌A549细胞系;利用北京博奥生物芯片公司的人类V2.0全基因组寡核苷酸微阵列芯片,检测其与正常贴壁生长A549细胞的基因表达差异性,筛选肺癌转移相关基因。结果共得到表达差异的基因745个,从中筛选出63个与肺癌转移密切相关的基因。结论成功建立抗脱落凋亡肺癌A549细胞系并筛选出转移相关差异表达基因,为进一步研究肺癌转移相关信号转导通路及其它相关研究提供依据。     

人膀胱移行细胞癌复发相关基因的筛选

目的：应用基因芯片技术筛选人膀胱移行细胞癌复发相关基因。方法：使用人肿瘤基因表达谱芯片检测11例膀胱移行细胞癌患者的正常膀胱粘膜组织、原发膀胱癌组织及复发膀胱癌组织的基因表达谱．筛选出在原发膀胱癌组织及复发膀胱癌组织中差异表达的基因，并用半定量RT—PCR对部分差异表达基因进行验证。结果：以正常膀胱粘膜组织为对照，11例膀胱肿瘤组织中有87个基因表达明显下调，102个基因表达明显上调．其中有15个基因的表达在原发膀胱癌组织及复发膀胱癌组织呈相反变化。结论：膀胱肿瘤与正常膀胱粘膜组织之间存在大量差异表达的基因，说明膀胱肿瘤的发生与发展是多种肿瘤相关基因表达失常或肿瘤抑制基因失活所致：在原发膀胱癌组织及复发膀胱癌组织中呈反向变化的15个基因可能与膀胱癌的复发有关。     

全基因组芯片筛查非小细胞肺癌组织多药耐药相关基因

背景与目的筛查与非小细胞肺癌多药耐药相关的基因,为非小细胞肺癌的个体化治疗提供理论依据。方法将手术切除的肺癌组织细胞进行原代培养。首先,采用MTT法检测诺维本、吉西他滨、多西他赛、紫杉醇及顺铂对非小细胞肺癌组织细胞的抑制率和敏感性。再利用全基因组芯片筛选人高度敏感组和耐药组间的差异表达基因。结果共筛选出差异表达基因212个,与耐药组相比,高度敏感组中上调基因168个,下调基因44个。结论利用全基因组芯片筛查出212个可能与非小细胞肺癌多药耐药相关的基因,用于指导临床个体化治疗。     

全反式维甲酸激活的人肺腺癌细胞基因的筛选

筛选人肺腺癌细胞中全反式维甲酸（ａｌｌｔｒａｎｓ－ｒｅｔｉｎｏｃｉａｃｉｄ,ＡＴＲＡ）激活表达的基因。方法以我室构建的ＡＴＲＡ诱导前后人肺腺癌细胞系ＧＬＣ－８２的ｃＤＮＡ文库Ｌｉｂ．ＧＬＣ－８２,Ｌｉｂ,ＧＬＣ－８２．ＲＡ１,Ｌｉｂ,ＧＬＣ－８２．ｒａ４ＦＪＴＦ     

Expression Profile of Genes Modulated by Aloe emodin in Human U87 Glioblastoma Cells

Glioblastoma, the most aggressive and malignant form of glioma, appears to be resistant to variouschemotherapeutic agents. Hence, approaches have been intensively investigated to targeti specific molecularpathways involved in glioblastoma development and progression. Aloe emodin is believed to modulate theexpression of several genes in cancer cells. We aimed to understand the molecular mechanisms underlying thetherapeutic effect of Aloe emodin on gene expression profiles in the human U87 glioblastoma cell line utilizingmicroarray technology. The gene expression analysis revealed that a total of 8,226 gene alterations out of 28,869genes were detected after treatment with 58.6 μg/ml for 24 hours. Out of this total, 34 genes demonstratedstatistically significant change (p<0.05) ranging from 1.07 to 1.87 fold. The results revealed that 22 genes wereup-regulated and 12 genes were down-regulated in response to Aloe emodin treatment. These genes were thengrouped into several clusters based on their biological functions, revealing induction of expression of genesinvolved in apoptosis (programmed cell death) and tissue remodelling in U87 cells (p<0.01). Several genes withsignificant changes of the expression level e.g. SHARPIN, BCAP31, FIS1, RAC1 and TGM2 from the apoptoticcluster were confirmed by quantitative real-time PCR (qRT-PCR). These results could serve as guidance forfurther studies in order to discover molecular targets for the cancer therapy based on Aloe emodin treatment.     

人白血病细胞系筛选抗癌药物的可行性

背景与目的 :探讨以人白血病细胞系筛选抗癌药物的可行性。 材料与方法 :应用活细胞计数法 ,3H_TdR及 3H_UR掺入 ,克隆分析法判断抗癌药物对肿癌细胞的杀伤能力。结果 :用人白血病细胞系K562和HL_60来检测对3种已知抗肿瘤药物(阿糖胞苷、柔红霉素、三尖杉酯碱)和一种新的药物MTB。在实验条件完全相同的条件下 ,同时用4种方法在不同药物浓度下存活分数曲线十分接近 ,其敏感范围在10 -2以内 ,克隆分析法检测敏感度可达10 -7水平。K562细胞存活分数高于HL_60细胞 ,与临床治疗结果相一致。结论 :采用人白血病细胞系为靶细胞进行抗癌药物的选择具有一定的优越性 ,不仅实验操作简便 ,而且与临床有较好的相关性。     

Screening and Verifying of Metastasis-associated Genes of Human Lung Cancer Cell Lines with Different Metastatic Potential

Background and Objective Lung cancer is the most lethal malignangy that threatens human health and lives nowadays in the world, The overall cure rate of lung cancer is only 13% -15%,     

Isolation of Novel Differentially Expressed Genes Related to Human Glioma Using CDNA Microarray and Characterizations of Two Novel Full-length Genes

Identification of the genes that are differentially expressed between brain tumor and normal brain tissues is important for understanding the molecular basis of these nerve system tumors and for defining possible targets for therapeutic intervention. This investigation is intended to obtain differentially expressed genes related to human glioma using cDNA microarray. Total RNA was extracted from human glioma specimens and normal brain tissues, and mRNA was obtained by oligotex chromatography. The cDNA microarray contains 4366 novel cDNA clones. The results of hybridization were scanned using computer system. Two genes selected from the results of cDNA microarray hybridization were subsequently analyzed by bio-informatic approach, Northern blot, in situ hybridization and radiation hybridization. We demonstrated that at a differentially expressed ration of two to three times, 15 cDNA clones were considered differentially expressed. Two novel full-length genes were selected for further investigation, one named human PKI gene (clone 436F11, GenBank^TM with accession number: AF225513) was over-expressed in normal brain tissues and the other named human ribosomal protein L14.22 gene (clone 507E08, GenBank^TM with accession number: AF329277) was over-expressed in gliomas. Furthermore, the 436F11 gene was located on 6q21–q23 between the D6S304 and D6S2156 markers, while the 507E08 gene was located between the D14S1066 and D14S265 markers. We realized that cDNA microarray technology can be successfully applied to identify differentially expressed genes in human glioma. This approach is superior to routine representational difference analysis, suppression subtractive hybridization and Northern blot for detection and isolation of differentially expressed genes in different tissues. At present, we have discovered two novel full-length genes related to human glioma and their characterizations have been partially clarified.