OBJECTIVE Eriocalyxin B(EriB)is a natural diterpenoid purified fromIsodoneriocalyxvar.laxiflora,traditional Chinese herbal medicines,possesses strong antileukemic activity with low toxicity.In murine t(8;21)leukemia m...OBJECTIVE Eriocalyxin B(EriB)is a natural diterpenoid purified fromIsodoneriocalyxvar.laxiflora,traditional Chinese herbal medicines,possesses strong antileukemic activity with low toxicity.In murine t(8;21)leukemia models,EriB remarkably prolong the survival time and decreased the xenograft tumor size by targeting AML1-ETO oncoprotein.In angiogenesis research by the highly vascularized chorioallantoic membrane(CAM)of the chicken embryo further confirm its antiangiogenic activity.Microarray offers a high efficient approach to study the gene expression profile treated by EriB,so as to provide systematic information about potential mechanisms of EriB curing AML.METHODS The t(8;21)AML cell line Kasumi-1 is most sensitive to EriB.Cells are treated with Eri-B(0.5μmol·L-1)and collected in 2,6,12 and 24h,respectively.Using human cDNA microarray,we investigate the changes of differential gene expression of Kasumi-1cells by time before and after Eri-B treatment.Meanwhile,the mRNA expression of TRAF2,DEDD2,BAG3,SAT1,IQGAP1,C-myc and GRB2 is detected by semi-quantitative RT-PCR and real-time quantitative PCR.RESULTS The genes regulated significantly are correlated with cell proliferation,apoptosis,cell circle,regulation of transcription,response to stimulies and metabolism.Regulation of TNFR mediated apoptotic signaling by EriB plays a key role to induce apoptosis and cell circle,involved NF-κB,Ras-MAPK,cAMP/PKA,PI3K/Akt,Cyt c/caspase and p53-Rb signal pathways.After detected by SqRT-PCR and real-time quantitative PCR,the mRNA expressions of BAG3,DEDD2,SAT1 and C-myc are significantly changed.CONCLUSION These findings describes the probable mechanisms involved and the value of EriB as a promising candidate targeting apoptosis cascade and cell circle in treatment of AML.展开更多
差异表达分析常被用于各种疾病标志物的筛选研究中,如传统的t检验、显著性分析(significance of microarrays,SAM)检验、偏最小二乘(pa least square,PLS)等方法。然而,这些方法主要是通过比较不同分组之间基因表达均值的差异筛选标记物...差异表达分析常被用于各种疾病标志物的筛选研究中,如传统的t检验、显著性分析(significance of microarrays,SAM)检验、偏最小二乘(pa least square,PLS)等方法。然而,这些方法主要是通过比较不同分组之间基因表达均值的差异筛选标记物,忽视了物质之间的相互调控关系,致使研究结果不够稳定或检验效率低。在组学研究中,由于基因调控和蛋白质的互相作用,很有可能在表达量上还没有呈现出明显差别时.展开更多
ObjectiveTo investigate the gene expression changes in normal and degeneration lumbar intervertebral disc in humans, providing information for clinical. MethodsThe PCR products of 4096 human genes were spotted onto a ...ObjectiveTo investigate the gene expression changes in normal and degeneration lumbar intervertebral disc in humans, providing information for clinical. MethodsThe PCR products of 4096 human genes were spotted onto a kind of chemical-material-coated-glass slides. The total RNAs were isolated from the tissues. Both the mRNAs from the degeneration and normal lumbar intervertebral disc in humans were reversely transcribed to the cDNAs, which used as the hybridization probes with the incorporations of fluorescent dUTP. The mixed probes were then hybridized to the cDNA microarray. After high-stringent washing, the cDNA microarray was scanned for the fluorescent signals and analyzed with computer image analysis. ResultsAmong the 4096 targets, there were 706 genes whose expression levels differed between the degeneration and normal lumbar intervertebral disc in all cases, comprising 298 up-regulated and 358 down-regulated ones. ConclusionDNA microarray technology is an effective technique in screening for differently expressed genes between the degeneration and normal lumbar intervertebral disc. Cell apoptosis plays an important role in the process of lumbar intervertebral disc degeneration.展开更多
Phototropism, the induction of carotenogenesis and reproductive structures, and resetting of the circadian rhythm are controlled by blue light. Trichoderma is used as a photomorphogenetic model due to its ability to c...Phototropism, the induction of carotenogenesis and reproductive structures, and resetting of the circadian rhythm are controlled by blue light. Trichoderma is used as a photomorphogenetic model due to its ability to conidiate upon exposure to light. In total darkness, T. atroviride grows indefinitely as a mycelium provided that nutrients are not limiting. However, nutrient deprivation and light trigger the conidiation process. A pulse of blue light given to a radially growing colony induces synchronous sporulation. A ring of conidiophores bearing green conidia is produced at what had been the colony perimeter at the time of the light pulse. All known responses to blue light in N. crassa are initiated by a couple of transcription factors encoded by the white-collar genes (wc -1 and wc-2). WC-1 and WC-2 bind to the promoters of light regulated genes to rapidly activate transcription in response to light. In T. atroviride the photolyase encoding gene phr1 undergoes fast transcriptional activation in response to light. The presence of putative WCC binding boxes in the promoter of phr1, suggested that light responses in Trichoderma could be under the control of white-collar homologues. We cloned two genes and demonstrated by gene replacement that both are essential for photoconidiation and photolyase gene expression. Therefore, they were named blue-light regulator one and two (blr1 and blr2). The BLR1 protein has all the characteristics of a blue-light photoreceptor. The generation of subtractive cDNA libraries allowed us to identify novel, BLR independent, light responses including the regulation of gene expression by blue-light. In addition, we recently initiated a Trichoderma ESTs sequencing project. Until now, we have sequenced above 3000 ESTs, from which we have obtained approximately 1800 unigenes. This unigene set was printed in microarrays and used to search for light induced genes. Twenty five clearly induced and around thirty repressed genes have been detected. Among this set we have found both blr dependent and independent blue light induced genes, strengthening our view of the existence of alternative light perception pathways. We also show the first evidence for the entry of Trichoderma into the conidiation process caused by mechanical injury, which remains unaltered in the mutants. Finally, an unprecedented crosstalk between light and glucose sensing was found involving the BLR1 and BLR2 proteins in the control of carbon deprivation induced conidiation.展开更多
基金The project supported by National Natural Science Foundation of China(30772637)Yunnan Provincial Science and Technology Department(2014IA033)
文摘OBJECTIVE Eriocalyxin B(EriB)is a natural diterpenoid purified fromIsodoneriocalyxvar.laxiflora,traditional Chinese herbal medicines,possesses strong antileukemic activity with low toxicity.In murine t(8;21)leukemia models,EriB remarkably prolong the survival time and decreased the xenograft tumor size by targeting AML1-ETO oncoprotein.In angiogenesis research by the highly vascularized chorioallantoic membrane(CAM)of the chicken embryo further confirm its antiangiogenic activity.Microarray offers a high efficient approach to study the gene expression profile treated by EriB,so as to provide systematic information about potential mechanisms of EriB curing AML.METHODS The t(8;21)AML cell line Kasumi-1 is most sensitive to EriB.Cells are treated with Eri-B(0.5μmol·L-1)and collected in 2,6,12 and 24h,respectively.Using human cDNA microarray,we investigate the changes of differential gene expression of Kasumi-1cells by time before and after Eri-B treatment.Meanwhile,the mRNA expression of TRAF2,DEDD2,BAG3,SAT1,IQGAP1,C-myc and GRB2 is detected by semi-quantitative RT-PCR and real-time quantitative PCR.RESULTS The genes regulated significantly are correlated with cell proliferation,apoptosis,cell circle,regulation of transcription,response to stimulies and metabolism.Regulation of TNFR mediated apoptotic signaling by EriB plays a key role to induce apoptosis and cell circle,involved NF-κB,Ras-MAPK,cAMP/PKA,PI3K/Akt,Cyt c/caspase and p53-Rb signal pathways.After detected by SqRT-PCR and real-time quantitative PCR,the mRNA expressions of BAG3,DEDD2,SAT1 and C-myc are significantly changed.CONCLUSION These findings describes the probable mechanisms involved and the value of EriB as a promising candidate targeting apoptosis cascade and cell circle in treatment of AML.
文摘差异表达分析常被用于各种疾病标志物的筛选研究中,如传统的t检验、显著性分析(significance of microarrays,SAM)检验、偏最小二乘(pa least square,PLS)等方法。然而,这些方法主要是通过比较不同分组之间基因表达均值的差异筛选标记物,忽视了物质之间的相互调控关系,致使研究结果不够稳定或检验效率低。在组学研究中,由于基因调控和蛋白质的互相作用,很有可能在表达量上还没有呈现出明显差别时.
文摘ObjectiveTo investigate the gene expression changes in normal and degeneration lumbar intervertebral disc in humans, providing information for clinical. MethodsThe PCR products of 4096 human genes were spotted onto a kind of chemical-material-coated-glass slides. The total RNAs were isolated from the tissues. Both the mRNAs from the degeneration and normal lumbar intervertebral disc in humans were reversely transcribed to the cDNAs, which used as the hybridization probes with the incorporations of fluorescent dUTP. The mixed probes were then hybridized to the cDNA microarray. After high-stringent washing, the cDNA microarray was scanned for the fluorescent signals and analyzed with computer image analysis. ResultsAmong the 4096 targets, there were 706 genes whose expression levels differed between the degeneration and normal lumbar intervertebral disc in all cases, comprising 298 up-regulated and 358 down-regulated ones. ConclusionDNA microarray technology is an effective technique in screening for differently expressed genes between the degeneration and normal lumbar intervertebral disc. Cell apoptosis plays an important role in the process of lumbar intervertebral disc degeneration.
文摘Phototropism, the induction of carotenogenesis and reproductive structures, and resetting of the circadian rhythm are controlled by blue light. Trichoderma is used as a photomorphogenetic model due to its ability to conidiate upon exposure to light. In total darkness, T. atroviride grows indefinitely as a mycelium provided that nutrients are not limiting. However, nutrient deprivation and light trigger the conidiation process. A pulse of blue light given to a radially growing colony induces synchronous sporulation. A ring of conidiophores bearing green conidia is produced at what had been the colony perimeter at the time of the light pulse. All known responses to blue light in N. crassa are initiated by a couple of transcription factors encoded by the white-collar genes (wc -1 and wc-2). WC-1 and WC-2 bind to the promoters of light regulated genes to rapidly activate transcription in response to light. In T. atroviride the photolyase encoding gene phr1 undergoes fast transcriptional activation in response to light. The presence of putative WCC binding boxes in the promoter of phr1, suggested that light responses in Trichoderma could be under the control of white-collar homologues. We cloned two genes and demonstrated by gene replacement that both are essential for photoconidiation and photolyase gene expression. Therefore, they were named blue-light regulator one and two (blr1 and blr2). The BLR1 protein has all the characteristics of a blue-light photoreceptor. The generation of subtractive cDNA libraries allowed us to identify novel, BLR independent, light responses including the regulation of gene expression by blue-light. In addition, we recently initiated a Trichoderma ESTs sequencing project. Until now, we have sequenced above 3000 ESTs, from which we have obtained approximately 1800 unigenes. This unigene set was printed in microarrays and used to search for light induced genes. Twenty five clearly induced and around thirty repressed genes have been detected. Among this set we have found both blr dependent and independent blue light induced genes, strengthening our view of the existence of alternative light perception pathways. We also show the first evidence for the entry of Trichoderma into the conidiation process caused by mechanical injury, which remains unaltered in the mutants. Finally, an unprecedented crosstalk between light and glucose sensing was found involving the BLR1 and BLR2 proteins in the control of carbon deprivation induced conidiation.
