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AIM: To analyze the lovastatin-induced differential gene expression in HepG2 cells using a cDNA microarray assay. METHODS: Total RNA was extracted from the lovastatin-treated HepG2 cells and control group. cDNA was synthesized from RNA with Cy3/Cy5-labelled dCTP. Then the hybridization was conducted. The result was analyzed using Imagene and Genespring software. RT-PCR was carried to confirm the hybridization results. RESULTS: 30 genes were up-regulated while 11 genes were down-regulated in lovastatin-treated HepG2 cells, involved in some major functional areas including signal transduction, cell cycle regulation, tumor immunity, and so on. CONCLUSION: The analysis of differentially expressed genes in lovastatin-treated HepG2 cells is helpful to explore the mechanism of the anti-tumor activity of statins.  相似文献   

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AIM:To investigate the effect of LY980503(a benflumetol derivative)on multidrug resistance of tumor cell line using DNA microarray.METHODS:Total RNA was extracted from multidrug resistant MCF/DOX cell line. cDNA microarray containing 320 cDNAs was used to detect the gene expression profile.RESULTS:9 down-regulated genes and 1 up-regulated gene were identified after multidrug resistant MCF/DOX cells were treated with LY980503.CONCLUSION:LY980503 can effectively reverse the resistance of MCF/DOX to DOX in vitro by adjusting the expression of multi-genes.  相似文献   

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梨S基因芯片的试制及分子杂交条件的优化   总被引:5,自引:0,他引:5  
 为研究寡核苷酸芯片在确定梨品种S基因型中的应用价值,根据梨自交不亲和基因的结构特点设计了部分S基因的检测探针,并制成寡核苷酸芯片;采用引物Cy3荧光标记法标记检测样品的PCR产物并进行杂交,以检测不同样品的S基因型。结果表明:通过对不同杂交温度、杂交时间等的探索获得了较好的反应条件并用制备的芯片检测了已知S基因型的梨样品,检测结果与各品种的已知基因型相符。证明寡核苷酸芯片检测梨品种的S基因型是一种切实可行的检测方法,若对芯片进一步完善,则今后在梨的自交不亲和性状的机理研究及梨自交不亲和性状的利用等方面将有着广阔的应用前景。  相似文献   

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AIM: To detect the differentially expressed genes associated with ovarian serous cystadenocarcinoma (OV) by microarray and to analyze the participated signaling pathway. METHODS: We analyzed 16 datasets of Affymetrix GeneChip Human Exon 1.0 ST Arrays from The Cancer Genome Atlas (TCGA), including 8 OV and 8 normal ovary samples. The function of differential genes was determined by pathway and gene ontology (GO) analysis. The probable functions of the key genes were predicted according to intergenic signal transduction network. RESULTS: The 1 144 genes were identified as distinctively expressed in OV (P<0.05), 747 of which were up-regulated and 397 were down-regulated. The GO analysis results showed that the altered genes were involved in 362 up-regulated and 160 down-regulated significant functions (P<0.05) related to cell cycle, DNA replication, cell proliferation, cell apoptosis, cell adhesion, etc. The pathways of the different genes were involved in the 59 enrichment-related pathways (P<0.05), 45 of which were up-regulated and 14 were down-regulated. Among the 59 pathways, cell cycle, P53 signaling pathway, DNA replication, pathways in cancer, PI3K-Akt signaling pathway, ECM-receptor signaling pathway, cell adhesion molecules and cell apoptosis were related to tumor genesis, development and metastasis. As a result, 229 genes with significant functions and pathways in GO and pathway analysis were selected to construct signal transduction network (Signal-Net), 4 of which, CDK1, PLK1, MCM3 and PGK1, were found to play key roles in OV signal regulation network. CONCLUSION: The OV shows abundant differentially expressed genes that play key roles in cancer-related signal pathways.  相似文献   

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AIM:To investigate the influence of ovariectomy and estrogen replacement treatment on profile of gene expression in myocardium by cDNA microarray,and to characterize the targeting genes of estrogen.METHODS:cDNA microarray containing 1 400 rat cDNAs was used to study the genes differentially expressed in myocardium between sham (Ⅰ),ovariectomy (Ⅱ,OVX) and estrogen replacement treatment (Ⅲ,OVX+E2) group.Then down-regulated genes in myocardium of OVX rats were further confirmed by RT-PCR.RESULTS:177 genes were differentially expressed in myocardium between sham and OVX rats,with 91 genes up-regulated and 86 genes down-regulated in OVX rats.164 genes were differentially expressed in myocardium between OVX and OVX+E2 rats,with 113 genes up-regulated and 54 genes down-regulated in OVX rats.There were 54 genes differentially expressed in OVX compared to sham and OVX+E2.They are involved in membrane channels and transporters (18),cell receptors (9),intracellular transducers/effectors/modulator (7) and metabolism (6).Most of the genes (45) were down-regulated in OVX rats and up-regulated in OVX+E2 rats.RT-PCR test confirmed the results of cDNA microarray.CONCLUSIONS:Long-term estrogen replacement may influence the expression of genes involved in membrane channels and transporters,cell receptors,intracellular transducers/effectors/ modulator and metabolism.Long-term estrogen replacement has some beneficial effects on ionic concentration and cardiac function which partially comes from the results of influence of expression on Na+,K+-ATPase and Na+/H+ exchanger.Estrogen has an inhibitory effect on the expression of dopamine receptor,which partially clarify the myocardial protection of estrogen.  相似文献   

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江南  谭晓风  张琳  邓靖 《园艺学报》2014,41(10):1983
根据东方梨中已鉴定的46个S基因序列和S基因的结构特点,设计了86条寡核苷酸探针并制备成S基因寡核苷酸检测芯片,采用Cy3荧光修饰引物标记被检测品种的PCR产物并与芯片杂交,以检测不同品种的S基因型。结果表明:利用芯片与华梨2号、秀玉和德胜香等已知S基因型的品种杂交,杂交信号显示与各品种已知基因型相符合。利用芯片鉴定了丽江黄酸梨等27个未知S基因型的梨品种,获得了各品种的S基因型,随机选取部分品种进行DNA测序和序列分析,结果与芯片杂交结果完全一致,证明利用S基因寡核苷酸芯片鉴定梨品种S基因型结果准确可靠。  相似文献   

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AIM: To screen the expression of inflammatory genes associated with atherosclerosis (AS) in different weeks of ApoE-/- mice using Agilent gene expression profile chip (AGEPC). METHODS: Male ApoE-/- mice (n=60) were randomly divided into 3 groups:initial phase of AS (10 weeks old), early phase of AS (15 weeks old), and late phase of AS (25 weeks old). Homologous wild-type C57BL/6J mice were used for the control. The RNA samples of the arcus aortae from these mice were isolated. Total RNA from each sample was labeled with Cy3 and hybridized with AGEPC, and microarray detection was conducted. After washing, scaning, acquiring data, and standardized analysis, the expressed genes with default threshold of statistical significance of P≤0.05 and fold change ≥ 2.0 were selected. The expression of these genes were further verified by RT-qPCR. RESULTS: Compared with the control group, there were 895 differential genes in 10 weeks of ApoE-/- mice, while 540 genes in 15 weeks, and 591 genes in 25 weeks, respectively. KEGG pathway and gene ontology (GO) analyses revealed that those diversely expressed genes related to inflammation were particularly arresting. Several selected genes including interleukin-12a (IL-12a), matrix metallopeptidase-12 (MMP-12), IL-1β, growth differentiation factor-15 (GDF-15) and interferon-γ (IFN-γ) were validated by RT-qPCR. Compared with the control group, the expression levels of IL-12a and MMP-12 were up-regulated while IL-1β was down-regulated in 10 weeks, the expression level of GDF-15 was up-regulated while the IL-12a and IL-1β levels were down-regulated in 15 weeks, and the levels of IL-12a, MMP-12 and GDF-15 were up-regulated in 25 weeks (P<0.05). Moreover, the increased level of IL-12a in 10 weeks, decreased level of IL-1β in 15 weeks, and increased levels of MMP-12 and GDF-15 in 25 weeks were even more statistically significant (P<0.01). CONCLUSION: The changes of inflammatory gene expression in different phases of AS suggest an important direction for medical intervention of AS.  相似文献   

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AIM:To explore the effect of pinobanksin-3-acetate (PB3A) on microRNA (miRNA) expression profile of human colon cancer cells for providing new methods of treatment of colon cancer and development of targeted drug.METHODS:The method of miRNA expression profiling was used to observe the miRNA differential expression in human colon cancer SW480 cells after treated with PB3A.The expression of miRNA-198 and miRNA-296-5p in the SW480 cells was detected by RT-qPCR.The network databases of miRWalk,MicroT,miRanda and so on were used to predict the target genes regulated by these miRNAs,and pathway significant enrichment analysis was performed.RESULTS:miRNA microarray analysis showed that after treated with propolis flavonoid PB3A for 24 h,267 miRNAs with differential expression twice or more in the SW480 cells were observed.Among them,there were 30 miRNAs with 10-fold or more differential expression,in which 28 were up-regulated and 2 were down-regulated.The results of RT-qPCR showed that the expression levels of miRNA-198 and miRNA-296-5p were consistent with the results of miRNA microarray analysis,and the difference was statistically significant (P<0.05).Bioinformatic analysis revealed that miRNA-198 has 859 target genes,and miRNA-296-5p has 906 target genes.The target genes of miRNA-198 were clustered in pathways in cancer,axon guidance,Wnt signaling pathway,regulation of actin cytoskeleton,insulin signaling pathway and MAPK signaling pathway,while the target genes of miRNA-296-5p were clustered in axon guidance,Wnt signaling pathway,MAPK signaling pathway,endocytosis,melanogenesis,insulin signaling pathway and calcium signaling pathway.CONCLUSION:Propolis flavonoid PB3A affects the expression of miRNA in colon cancer SW480 cells.The abnormal expression of miRNA-198 and miRNA-296-5p may be involved in the inhibitory effect of PB3A on colon cancer.  相似文献   

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江南    谭晓风  张琳  张靖国  胡红菊 《园艺学报》2015,42(12):2341-2352
利用东方梨中已鉴定的52个S等位基因HV区cDNA序列作为靶基因序列设计探针,制备梨S基因cDNA检测芯片,每张芯片上含有240个位点55个cDNA探针,包含所有序列完善的S基因HV区特异的cDNA序列。以被检测品种雌蕊cDNA为模板,采用Cy3荧光修饰引物经S基因特异PCR扩增标记被检测品种的cDNA序列,并与芯片杂交以检测不同品种的S基因型。结果表明:利用cDNA检测芯片与‘丽江黄酸梨’、‘秀玉’、‘弥渡玉梨’、‘白面梨’和‘德胜香’等已知S基因型品种杂交,杂交结果显示与S基因寡核苷酸芯片检测信号一致,与各品种已知S基因型相符合。利用cDNA芯片和进一步完善的S基因寡核苷酸芯片并行检测鉴定了‘文山红梨’等24个未知S基因型的砂梨品种,获得各品种的S基因型。梨S基因cDNA芯片的构建进一步完善了梨S基因检测平台。  相似文献   

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AIM:To examine the expression profiles of both genes and proteins in hippocampus of rats with temporal lobe epilepsy (TLE) for revealing the molecular mechanisms of TLE and looking for the candidate targets and new therapeutic approaches in clinical practice.METHODS:Rat temporal lobe epilepsy was induced by administration of lithium chloride and pilocarpine (LiCl-PILO).The expression spectra of genes and proteins were constructed through the techniques of cDNA microarray,two-dimensional (2D) electrophoresis and Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).Subsequently,the differentially expressed genes and proteins were identified and analyzed.RESULTS:There were 192 genes of differential expression observed in hippocampal tissues of LiCl-PILO-induced temporal lobe epilepsy,and 159 genes have been registered in Genbank database,in which 84 genes were up-regulated while 75 genes were down-regulated.78 protein spots of differential display were screened out,in which 31 proteins were detected to be down-regulated and 47 were up-regulated.Finally,5 proteins were identified.CONCLUSION:These genes and proteins found in our study may play pivotal roles in the pathogenic mechanisms of epilepsy and may promise new therapeutic targets for refractory epilepsy in the future.  相似文献   

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AIM: To investigate the effects of exosomes secreted by pancreatic cancer cells on the viability and function of β cells and the possible mechanism. METHODS: ExoQuick-TC kit was used to extract exosomes in the supernatants of mouse pancreatic cancer Pan02 and MPC-83 cells, and the extracted exosomes were identified by transmission electron microscopy. Fluorescence-labeled exosomes were incubated with mouse insulinoma MIN6 cells for 48 h to detect whether exosomes secreted by pancreatic cancer cells were uptaken by MIN6 cells. MTT and glucose-stimulated insulin secretion (GSIS) assays were conducted to examine cell viability and insulin secretion of MIN6 cells after incubating with exosomes. The expression of miR-204 and Bcl-2 mRNA in MIN6 cells was detected by qPCR. The protein expression of Bcl-2, Bax, caspase-3 and cytochrome C (Cyt-C) in MIN6 cells was determined by Western blot. RESULTS: The results of transmission electron microscopy showed that both Pan02 cells and MPC-83 cells secreted exosomes, and Pan02 cells secreted more. The co-incubation results of fluorescence-labeled exosomes and MIN6 cells confirmed that MIN6 cells were able to ingest large amounts of exosomes secreted by pancreatic cancer cells. The results of MTT and GSIS assays showed that the viability and the level of high glucose-stimulated insulin secretion of MIN6 cells in exosome treatment group significantly decreased compared with nontreatment group (P<0.01). The results of qPCR showed that the exosomes secreted by pancreatic cancer cells were rich in miR-204, and the mRNA expression of Bcl-2 in MIN6 cells was significantly down-regulated by exosome incubation (P<0.01). The results of Western blot showed that the protein expression of Bcl-2 in the MIN6 cells treated with exosomes was significantly down-regulated (P<0.05), and the protein levels of Bax, cleaved caspase-3 and Cyt-C in exosomes treatment group were significantly up-regulated (P<0.01). CONCLUSION: Pancreatic cancer cells secrete exosomes. The exosomes secreted by pancreatic cancer cells are ingested by β cells, and reduce the viability and insulin secretion of β cells. The mechanism may be related to the increase in exosomal miR-204 in the β cells. Increasing miR-204 may inhibit the expression of Bcl-2 and promote the activation of mitochondrial apoptosis in β cells.  相似文献   

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