The highbush blueberry（Vaccinium corymbosum）,Duke,was used to construct a de novo transcriptome sequence library and to perform data statistical analysis.Mega 4,CLC Sequence Viewer 6 software,and quantitative PCR we...The highbush blueberry（Vaccinium corymbosum）,Duke,was used to construct a de novo transcriptome sequence library and to perform data statistical analysis.Mega 4,CLC Sequence Viewer 6 software,and quantitative PCR were employed for bioinformatics and expression analyses of the basic helix-loop-helix（BHLH）transcription factors of the sequencing library.The results showed that 28.38 gigabytes of valid data were obtained from transcriptome sequencing and were assembled into 108 033 unigenes.Functional annotation showed that 32 244 unigenes were annotated into Clusters of Orthologous Groups（COG）and Gene Ontology（GO）databases,whereas the rest of the 75 789 unigenes had no matching information.By using COG and GO classification tools,sequences with annotation information were divided into 25 and 52 categories,respectively,which involved transport and metabolism,transcriptional regulation,and signal transduction.Analysis of the transcriptome library identified a total of 59 BHLH genes.Sequence analysis revealed that 55 genes of that contained a complete BHLH domain.Furthermore,phylogenetic analysis showed that BHLH genes of blueberry（Duke）could be divided into 13 sub-groups.PCR results showed that 45 genes were expressed at various developmental stages of buds,stems,leaves,flowers,and fruits,suggesting that the function of BHLH was associated with the development of different tissues and organs of blueberry,Duke.The present study would provided a foundation for further investigations on the classification and functions of the blueberry BHLH family.展开更多
Basic helix-loop-helix （bHLH） transcription factors regulate the differentiation of various tissues in a vast diversity of species. The bHLH protein Atonal was first identified as a proneural gene involved in the fo...Basic helix-loop-helix （bHLH） transcription factors regulate the differentiation of various tissues in a vast diversity of species. The bHLH protein Atonal was first identified as a proneural gene involved in the formation of mechanosensory cells and photoreceptor cells in Drosophila （larman et al., 1993, 1994）. Atonal is expressed in sensory organ precursors and is required and sufficient for the development of chordotonal organs （Jar- man et al., 1993）. Moreover, Atonal expression is observed in the developing eye and is essential for the differentiation of R8 photoreceptors, which are the first photoreceptors that appear during development. Atonal is not involved in the formation of other photoreceptors （R1-R7） directly. However, R8 photore- ceptors recruit other photoreceptors from the surrounding cells （Jarman et al., 1994）.展开更多
Cell cycle progression is regulated by interactions between cyclins and cyclin-dependent kinases (CDKs). p21(WAF1) is one of the CIP/KIP family which inhibits CDKs activity. Increased expression of p21(WAF1) may play ...Cell cycle progression is regulated by interactions between cyclins and cyclin-dependent kinases (CDKs). p21(WAF1) is one of the CIP/KIP family which inhibits CDKs activity. Increased expression of p21(WAF1) may play an important role in the growth arrest induced in transformed cells. Although the stability of the p21( WAF1) mRNA could be altered by different signals, cell differentiation and numerous influencing factors. However, recent studies suggest that two known mechanisms of epigenesis, i.e.gene inactivation by methylation in promoter region and changes to an inactive chromatin by histone deacetylation, seem to be the best candidate mechanisms for inactivation of p21( WAF1). To date, almost no coding region p21(WAF1) mutations have been found in tumor cells, despite extensive screening of hundreds of various tumors. Hypermethylation of the p21(WAF1) promoter region may represent an alternative mechanism by which the p21(WAF1/CIP1) gene can be inactivated. The reduction of cellular DNMT protein levels also induces a corresponding rapid increase in the cell cycle regulator p21(WAF1) protein demonstrating a regulatory link between DNMT and p21(WAF1) which is independent of methylation of DNA. Both histone hyperacetylation and hypoacetylation appear to be important in the carcinoma process, and induction of the p21(WAF1) gene by histone hyperacetylation may be a mechanism by which dietary fiber prevents carcinogenesis. Here, we review the influence of histone acetylation and DNA methylation on p21(WAF1) transcription, and affection of pathways or factors associated such as p 53, E2A, Sp1 as well as several histone deacetylation inhibitors.展开更多
Background Transcription factors hypoxia inducible factor 1α （HIF 1α） and endothelial PAS domain protein 1 （EPAS1） promote the transcription of vascular endothelial growth factor （VEGF）. VEGF enhances angiogen...Background Transcription factors hypoxia inducible factor 1α （HIF 1α） and endothelial PAS domain protein 1 （EPAS1） promote the transcription of vascular endothelial growth factor （VEGF）. VEGF enhances angiogenesis and vascular permeability of tumours, which promotes tumour growth and facilitates entry of cancer cells into blood circulation and metastasizing. This study examined whether HIF 1α and EPAS1 stimulated angiogenesis through activation of VEGF in human pancreatic carcinoma. Methods Specimens from pancreatic carcinoma and healthy parts of same pancreas were taken from 60 patients. Real time quantitative reverse transcription polymerase chain reaction estimated expression of HIF 1α, EPAS1, and VEGF mRNAs. Western blotting and immunohistochemical, streptavidin peroxidase method assessed expression of HIF 1α, EPAS1, and VEGF proteins. Microvessel density （MVD） was assessed. Results Highly significant increases in expression of EPAS1, VEGF, and MVD were found in pancreatic carcinoma tissue but not in normal pancreatic tissue： VEGF at mRNA and protein levels （t=17.32, P=-0.0001; t=98.41, P=0.0001）; EPAS1 protein level （t=22.51, P=0.0001）. Expression of HIF la was similar in pancreatic carcinoma and normal pancreatic tissues at both mRNA and protein levels. Significant correlations were observed between EPAS1 and VEGF （r=0.736, P=0.0041）, between VEGF and MVD （r=0.858, P=0.0001）, and between EPAS1 and MVD （r=0.641, P=0.0003）. No significant correlations were observed between HIF la and VEGF, or between HIF 1α and MVD. MVD and expression of EPAS1 and VEGF were significantly related with TNM staging, so was EPASI and VEGF with size of tumour. Conclusions EPAS1 and VEGF, but not HIFla, are overexpressed in pancreatic carcinoma. The expression of EPAS1 is correlated with that of VEGF and MVD. EPAS1 may be involved in the angiogenesis of pancreatic carcinoma by upregulating the expression of VEGE Targeting EPAS1 may be a new method of antiangiogenic tumour therapy for pancreatic carcinoma.展开更多
Oligodencrocytes（OLs） are the main glial cells of the central nervous system involved in myelination of axons. In multiple sclerosis（MS）, there is an imbalance between demyelination and remyelination processes, th...Oligodencrocytes（OLs） are the main glial cells of the central nervous system involved in myelination of axons. In multiple sclerosis（MS）, there is an imbalance between demyelination and remyelination processes, the last one performed by oligodendrocyte progenitor cells（OPCs） and OLs, resulting into a permanent demyelination, axonal damage and neuronal loss. In MS lesions, astrocytes and microglias play an important part in permeabilization of blood-brain barrier and initiation of OPCs proliferation. Migration and differentiation of OPCs are influenced by various factors and the process is finalized by insufficient acummulation of OLs into the MS lesion. In relation to all these processes, the author will discuss the potential targets for remyelination strategies.展开更多
目的探讨miR-1271对卵巢癌细胞的增殖、侵袭和迁移的影响,及其作用机制。方法实验分为miR-NC组(转染miR-NC)、miR-1271组(转染miR-1271 mimics)、miR-1271+pcDNA3.1组(共转染miR-1271和pcDNA3.1)、miR-1271+pcDNA3.1-Twist1组(共转染miR...目的探讨miR-1271对卵巢癌细胞的增殖、侵袭和迁移的影响,及其作用机制。方法实验分为miR-NC组(转染miR-NC)、miR-1271组(转染miR-1271 mimics)、miR-1271+pcDNA3.1组(共转染miR-1271和pcDNA3.1)、miR-1271+pcDNA3.1-Twist1组(共转染miR-1271和pcDNA3.1-Twist1)。用噻唑蓝法检测细胞活性,用Transwell检测细胞迁移和侵袭情况,用实时定量聚合酶链反应检测miR-1271和碱性螺旋-环-螺旋转录因子1(Twist1)mRNA的表达水平,用荧光素酶报告基因检测实验检测miR-1271对Twist1的靶向调控。结果干预24 h后,miR-NC组、miR-1271组、miR-1271+pcDNA3.1组和miR-1271+pcDNA3.1-Twist1组的细胞活性分别为0.71±0.07,0.46±0.04,0.42±0.04和0.57±0.05,迁移细胞数分别为(129.00±9.69),(68.00±3.26),(67.00±3.18)和(98.00±6.85)个,侵袭细胞数分别为(110.00±8.16),(53.00±2.91),(55.00±2.96)和(83.00±4.39)个,miR-1271表达水平分别为0.18±0.02,0.47±0.04,0.46±0.06和0.49±0.01,Twist1 mRNA表达水平分别为0.68±0.06,0.13±0.01,0.11±0.01和0.47±0.03。miR-1271组和miR-1271+pcDNA3.1-Twist1组的细胞活性、迁移和侵袭细胞数、miR-1271和Twist1 mRNA表达水平分别与miR-NC组和miR-1271+pcDNA3.1组比较,差异均有统计学意义(均P<0.05)。转染WT-Twist1后,相较于miR-NC组,miR-1271组的荧光素酶活性显著降低(0.63±0.06 vs 1.12±0.10,P<0.05);而转染MUT-Twist1后,相较于miR-con组,miR-1271组的荧光素酶活性无显著变化(1.09±0.10 vs 1.11±0.10,P>0.05)。结论miR-1271可抑制卵巢癌细胞的增殖、迁移和侵袭,其机制可能与Twist1有关。展开更多
Pod shattering is an agronomical trait that is a result of the coordinated action of cell differentiation and separation. In Arabidopsis, pod shattering is controlled by a complex genetic network in which ALCATRAZ （...Pod shattering is an agronomical trait that is a result of the coordinated action of cell differentiation and separation. In Arabidopsis, pod shattering is controlled by a complex genetic network in which ALCATRAZ （ALC）, a member of the basic helix-loop-helix family, is critical for cell separation during fruit dehiscence. Herein, we report the identification of ALC-INTERACTING PROTEIN1 （ACI1） via the yeast two-hybrid screen. ACll encodes a nuclear protein with a lysine-rich domain and a C-terminal serine-rich domain. ACI1 is mainly expressed in the vascular system throughout the plant and mesocarp of the valve in siliques. Our data showed that ACI1 interacts strongly with the N-terminal portion of ALC in yeast cells and in plant cells in the nucleus as demonstrated by bimolecular fluorescence complementaUon assay. Both ACI1 and ALC share an overlapping expression pattern, suggesting that they likely function together in planta. However, no detectable phenotype was found in plants with reduced ACll expression by RNA interference technology, suggesting that ACll may be redundant. Taken together, these data indicate that ALC may interact with ACI1 and its homologs to control cell separation during fruit dehiscence in Arabidopsis.展开更多
Jasmonic acid(JA)is a key regulator of plant defense responses.Although the transcription factor MYC2,the master regulator of the JA signaling pathway,orchestrates a hierarchical transcriptional cascade that regulates...Jasmonic acid(JA)is a key regulator of plant defense responses.Although the transcription factor MYC2,the master regulator of the JA signaling pathway,orchestrates a hierarchical transcriptional cascade that regulates the JA responses,only a few transcriptional regulators involved in this cascade have been described.Here,we identified the basic helix-loop-helix(bHLH)transcription factor gene in tomato(Solanum lycopersicum),METHYL JASMONATE(MeJA)-INDUCED GENE(SlJIG),the expression of which was strongly induced by MeJA treatment.Genetic and molecular biology experiments revealed that SlJIG is a direct target of MYC2.SlJIG knockout plants generated by gene editing had lower terpene contents than the wild type from the lower expression of TERPENE SYNTHASE(TPS)genes,rendering them more appealing to cotton bollworm(Helicoverpa armigera).Moreover,SlJIG knockouts exhibited weaker JA-mediated induction of TPSs,suggesting that SlJIG may participate in JA-induced terpene biosynthesis.Knocking out SlJIG also resulted in attenuated expression of JA-responsive defense genes,which may contribute to the observed lower resistance to cotton bollworm and to the fungus Botrytis cinerea.We conclude that SlJIG is a direct target of MYC2,forms a MYC2-SlJIG module,and functions in terpene biosynthesis and resistance against cotton bollworm and B.cinerea.展开更多
基金supported by the National Natural Science Foundation of China (31301754)the Chinese Academy of Agricultural Sciences-Agricultural Science and Technology Innovation Program (CAAS-ASTIP)the Cultivation Plan for Youth Agricultural Science and Technology Innovative Talents of Liaoning Province, China (2015059)
文摘The highbush blueberry（Vaccinium corymbosum）,Duke,was used to construct a de novo transcriptome sequence library and to perform data statistical analysis.Mega 4,CLC Sequence Viewer 6 software,and quantitative PCR were employed for bioinformatics and expression analyses of the basic helix-loop-helix（BHLH）transcription factors of the sequencing library.The results showed that 28.38 gigabytes of valid data were obtained from transcriptome sequencing and were assembled into 108 033 unigenes.Functional annotation showed that 32 244 unigenes were annotated into Clusters of Orthologous Groups（COG）and Gene Ontology（GO）databases,whereas the rest of the 75 789 unigenes had no matching information.By using COG and GO classification tools,sequences with annotation information were divided into 25 and 52 categories,respectively,which involved transport and metabolism,transcriptional regulation,and signal transduction.Analysis of the transcriptome library identified a total of 59 BHLH genes.Sequence analysis revealed that 55 genes of that contained a complete BHLH domain.Furthermore,phylogenetic analysis showed that BHLH genes of blueberry（Duke）could be divided into 13 sub-groups.PCR results showed that 45 genes were expressed at various developmental stages of buds,stems,leaves,flowers,and fruits,suggesting that the function of BHLH was associated with the development of different tissues and organs of blueberry,Duke.The present study would provided a foundation for further investigations on the classification and functions of the blueberry BHLH family.
基金supported by grants from the Ministry of Education,Culture,Sports,Science and Technology in Japan and Naito Foundation to TCthe Japan Society for the Promotion of Science to MO and TC
文摘Basic helix-loop-helix （bHLH） transcription factors regulate the differentiation of various tissues in a vast diversity of species. The bHLH protein Atonal was first identified as a proneural gene involved in the formation of mechanosensory cells and photoreceptor cells in Drosophila （larman et al., 1993, 1994）. Atonal is expressed in sensory organ precursors and is required and sufficient for the development of chordotonal organs （Jar- man et al., 1993）. Moreover, Atonal expression is observed in the developing eye and is essential for the differentiation of R8 photoreceptors, which are the first photoreceptors that appear during development. Atonal is not involved in the formation of other photoreceptors （R1-R7） directly. However, R8 photore- ceptors recruit other photoreceptors from the surrounding cells （Jarman et al., 1994）.
文摘Cell cycle progression is regulated by interactions between cyclins and cyclin-dependent kinases (CDKs). p21(WAF1) is one of the CIP/KIP family which inhibits CDKs activity. Increased expression of p21(WAF1) may play an important role in the growth arrest induced in transformed cells. Although the stability of the p21( WAF1) mRNA could be altered by different signals, cell differentiation and numerous influencing factors. However, recent studies suggest that two known mechanisms of epigenesis, i.e.gene inactivation by methylation in promoter region and changes to an inactive chromatin by histone deacetylation, seem to be the best candidate mechanisms for inactivation of p21( WAF1). To date, almost no coding region p21(WAF1) mutations have been found in tumor cells, despite extensive screening of hundreds of various tumors. Hypermethylation of the p21(WAF1) promoter region may represent an alternative mechanism by which the p21(WAF1/CIP1) gene can be inactivated. The reduction of cellular DNMT protein levels also induces a corresponding rapid increase in the cell cycle regulator p21(WAF1) protein demonstrating a regulatory link between DNMT and p21(WAF1) which is independent of methylation of DNA. Both histone hyperacetylation and hypoacetylation appear to be important in the carcinoma process, and induction of the p21(WAF1) gene by histone hyperacetylation may be a mechanism by which dietary fiber prevents carcinogenesis. Here, we review the influence of histone acetylation and DNA methylation on p21(WAF1) transcription, and affection of pathways or factors associated such as p 53, E2A, Sp1 as well as several histone deacetylation inhibitors.
文摘Background Transcription factors hypoxia inducible factor 1α （HIF 1α） and endothelial PAS domain protein 1 （EPAS1） promote the transcription of vascular endothelial growth factor （VEGF）. VEGF enhances angiogenesis and vascular permeability of tumours, which promotes tumour growth and facilitates entry of cancer cells into blood circulation and metastasizing. This study examined whether HIF 1α and EPAS1 stimulated angiogenesis through activation of VEGF in human pancreatic carcinoma. Methods Specimens from pancreatic carcinoma and healthy parts of same pancreas were taken from 60 patients. Real time quantitative reverse transcription polymerase chain reaction estimated expression of HIF 1α, EPAS1, and VEGF mRNAs. Western blotting and immunohistochemical, streptavidin peroxidase method assessed expression of HIF 1α, EPAS1, and VEGF proteins. Microvessel density （MVD） was assessed. Results Highly significant increases in expression of EPAS1, VEGF, and MVD were found in pancreatic carcinoma tissue but not in normal pancreatic tissue： VEGF at mRNA and protein levels （t=17.32, P=-0.0001; t=98.41, P=0.0001）; EPAS1 protein level （t=22.51, P=0.0001）. Expression of HIF la was similar in pancreatic carcinoma and normal pancreatic tissues at both mRNA and protein levels. Significant correlations were observed between EPAS1 and VEGF （r=0.736, P=0.0041）, between VEGF and MVD （r=0.858, P=0.0001）, and between EPAS1 and MVD （r=0.641, P=0.0003）. No significant correlations were observed between HIF la and VEGF, or between HIF 1α and MVD. MVD and expression of EPAS1 and VEGF were significantly related with TNM staging, so was EPASI and VEGF with size of tumour. Conclusions EPAS1 and VEGF, but not HIFla, are overexpressed in pancreatic carcinoma. The expression of EPAS1 is correlated with that of VEGF and MVD. EPAS1 may be involved in the angiogenesis of pancreatic carcinoma by upregulating the expression of VEGE Targeting EPAS1 may be a new method of antiangiogenic tumour therapy for pancreatic carcinoma.
文摘Oligodencrocytes（OLs） are the main glial cells of the central nervous system involved in myelination of axons. In multiple sclerosis（MS）, there is an imbalance between demyelination and remyelination processes, the last one performed by oligodendrocyte progenitor cells（OPCs） and OLs, resulting into a permanent demyelination, axonal damage and neuronal loss. In MS lesions, astrocytes and microglias play an important part in permeabilization of blood-brain barrier and initiation of OPCs proliferation. Migration and differentiation of OPCs are influenced by various factors and the process is finalized by insufficient acummulation of OLs into the MS lesion. In relation to all these processes, the author will discuss the potential targets for remyelination strategies.
文摘目的探讨miR-1271对卵巢癌细胞的增殖、侵袭和迁移的影响,及其作用机制。方法实验分为miR-NC组(转染miR-NC)、miR-1271组(转染miR-1271 mimics)、miR-1271+pcDNA3.1组(共转染miR-1271和pcDNA3.1)、miR-1271+pcDNA3.1-Twist1组(共转染miR-1271和pcDNA3.1-Twist1)。用噻唑蓝法检测细胞活性,用Transwell检测细胞迁移和侵袭情况,用实时定量聚合酶链反应检测miR-1271和碱性螺旋-环-螺旋转录因子1(Twist1)mRNA的表达水平,用荧光素酶报告基因检测实验检测miR-1271对Twist1的靶向调控。结果干预24 h后,miR-NC组、miR-1271组、miR-1271+pcDNA3.1组和miR-1271+pcDNA3.1-Twist1组的细胞活性分别为0.71±0.07,0.46±0.04,0.42±0.04和0.57±0.05,迁移细胞数分别为(129.00±9.69),(68.00±3.26),(67.00±3.18)和(98.00±6.85)个,侵袭细胞数分别为(110.00±8.16),(53.00±2.91),(55.00±2.96)和(83.00±4.39)个,miR-1271表达水平分别为0.18±0.02,0.47±0.04,0.46±0.06和0.49±0.01,Twist1 mRNA表达水平分别为0.68±0.06,0.13±0.01,0.11±0.01和0.47±0.03。miR-1271组和miR-1271+pcDNA3.1-Twist1组的细胞活性、迁移和侵袭细胞数、miR-1271和Twist1 mRNA表达水平分别与miR-NC组和miR-1271+pcDNA3.1组比较,差异均有统计学意义(均P<0.05)。转染WT-Twist1后,相较于miR-NC组,miR-1271组的荧光素酶活性显著降低(0.63±0.06 vs 1.12±0.10,P<0.05);而转染MUT-Twist1后,相较于miR-con组,miR-1271组的荧光素酶活性无显著变化(1.09±0.10 vs 1.11±0.10,P>0.05)。结论miR-1271可抑制卵巢癌细胞的增殖、迁移和侵袭,其机制可能与Twist1有关。
基金a Grant from the Ministry of Science and Technology of Chinato W.C. Yang (JY03-A-24)
文摘Pod shattering is an agronomical trait that is a result of the coordinated action of cell differentiation and separation. In Arabidopsis, pod shattering is controlled by a complex genetic network in which ALCATRAZ （ALC）, a member of the basic helix-loop-helix family, is critical for cell separation during fruit dehiscence. Herein, we report the identification of ALC-INTERACTING PROTEIN1 （ACI1） via the yeast two-hybrid screen. ACll encodes a nuclear protein with a lysine-rich domain and a C-terminal serine-rich domain. ACI1 is mainly expressed in the vascular system throughout the plant and mesocarp of the valve in siliques. Our data showed that ACI1 interacts strongly with the N-terminal portion of ALC in yeast cells and in plant cells in the nucleus as demonstrated by bimolecular fluorescence complementaUon assay. Both ACI1 and ALC share an overlapping expression pattern, suggesting that they likely function together in planta. However, no detectable phenotype was found in plants with reduced ACll expression by RNA interference technology, suggesting that ACll may be redundant. Taken together, these data indicate that ALC may interact with ACI1 and its homologs to control cell separation during fruit dehiscence in Arabidopsis.
基金the support from Beijing Modern Agricultural Industry System for facility vegetable cropsthe support from Beijing Key Laboratory of Growth and Developmental Regulation for Protected Vegetable Crops and Engineering Research Center of Breeding and Propagation of Horticultural Crops+2 种基金supported by grants from the National Natural Science Foundation of China(32002053,32172599)Hainan Provincial Joint Project of Sanya Yazhou Bay Science and Technology City(320LH013)the Administration of Sanya Yazhouwan Technology City(SYND-2022-07)and LJNY201812。
文摘Jasmonic acid(JA)is a key regulator of plant defense responses.Although the transcription factor MYC2,the master regulator of the JA signaling pathway,orchestrates a hierarchical transcriptional cascade that regulates the JA responses,only a few transcriptional regulators involved in this cascade have been described.Here,we identified the basic helix-loop-helix(bHLH)transcription factor gene in tomato(Solanum lycopersicum),METHYL JASMONATE(MeJA)-INDUCED GENE(SlJIG),the expression of which was strongly induced by MeJA treatment.Genetic and molecular biology experiments revealed that SlJIG is a direct target of MYC2.SlJIG knockout plants generated by gene editing had lower terpene contents than the wild type from the lower expression of TERPENE SYNTHASE(TPS)genes,rendering them more appealing to cotton bollworm(Helicoverpa armigera).Moreover,SlJIG knockouts exhibited weaker JA-mediated induction of TPSs,suggesting that SlJIG may participate in JA-induced terpene biosynthesis.Knocking out SlJIG also resulted in attenuated expression of JA-responsive defense genes,which may contribute to the observed lower resistance to cotton bollworm and to the fungus Botrytis cinerea.We conclude that SlJIG is a direct target of MYC2,forms a MYC2-SlJIG module,and functions in terpene biosynthesis and resistance against cotton bollworm and B.cinerea.
