目的:探讨Ig样结构域2黏附分子(adhesion molecule with Ig like domain 2,AMIGO2)在鼻咽癌(nasopharyngeal carcinoma,NPC)细胞增殖中的作用及其机制。方法:选用2017年9月至11月福建省肿瘤医院收集的10例NPC组织和10例正常鼻咽黏膜上...目的:探讨Ig样结构域2黏附分子(adhesion molecule with Ig like domain 2,AMIGO2)在鼻咽癌(nasopharyngeal carcinoma,NPC)细胞增殖中的作用及其机制。方法:选用2017年9月至11月福建省肿瘤医院收集的10例NPC组织和10例正常鼻咽黏膜上皮组织标本,以及NPC细胞系CNE-1、CNE-2、SUNE-1、6-10B、C666-1和人永生化鼻咽黏膜上皮细胞株NP69,用qPCR法检测NPC组织和细胞中AMIGO2 mRNA的表达。构建慢病毒载体干扰AMIGO2表达,用qPCR法验证其干扰效率;用CCK-8法、克隆形成及流式细胞术检测干扰AMIGO2表达对NPC细胞增殖、克隆形成和凋亡的影响,用Western blotting检测干扰AMIGO2表达对NPC细胞增殖及PI3K/AKT/mTOR信号通路相关标志蛋白表达的影响。结果:AMIGO2在NPC组织和CNE-2和SUNE-1细胞中高表达(均P<0.01)。慢病毒AMIGO2感染后,CNE-2和SUNE-1细胞的AMIGO2干扰效率均达50%以上。干扰AMIGO2表达,显著降低CNE-2和SUNE-1细胞增殖及克隆形成能力(均P<0.01)、明显提高细胞的凋亡率(均P<0.01);降低SUNE-1细胞中PI3K、AKT和mTOR磷酸化蛋白的表达水平(均P<0.01)、下调survivin和PCNA蛋白的表达水平(均P<0.01)。结论:AMIGO2通过激活PI3K/AKT/mTOR信号通路促进NPC细胞增殖并抑制其凋亡,提示AMIGO2可能是NPC治疗的潜在靶点。展开更多
Endophytic species of Colletotrichum associated with Mangifera indica(mango)are poorly understood.In this study,Colletotrichum species were isolated from mango in Pernambuco State,Brazil.There were significant differe...Endophytic species of Colletotrichum associated with Mangifera indica(mango)are poorly understood.In this study,Colletotrichum species were isolated from mango in Pernambuco State,Brazil.There were significant differences in isolation frequencies of Colletotrichum species among sites and plant tissues.Mature leaf blades were colonized by most Colletotrichum isolates at the majority of sites.Partial sequences of glyceraldehyde-3-phosphate dehydrogenase(GAPDH)of 97 Colletotrichum isolates were amplified as an initial measure of genetic diversity.Phylogenetic analysis with a subset of 22 isolates were performed based on a multilocus dataset(ACT,TUB2,CAL,CHS-1,GAPDH,ITS)followed by Apn2/MAT IGS sequence-analysis for isolates within the C.gloeosporioides species complex.Molecular analysis associated with phenotypic characteristics revealed six previously described species[C.asianum,C.cliviae,C.dianesei(syn.C.melanocaulon),C.fructicola,C.karstii and C.tropicale]and one new species.This new species is introduced as C.endomangiferae.All species isolated were pathogenic on mango fruits but varied in their virulence.There was no distribution pattern of species among sites and plant tissues,although C.asianum was the most prevalent species at all sites and in all plant tissues studied.Five previously reported Colletotrichum species causing anthracnose in mango fruits in northeastern Brazil were also recovered as endophytes.展开更多
Colletotrichum gloeosporioides sensu lato has been associated with anthracnose in diverse commercial crops.It is now established that C.gloeosporioides sensu lato comprises 33 phylogenetic species and C.gloeosporioide...Colletotrichum gloeosporioides sensu lato has been associated with anthracnose in diverse commercial crops.It is now established that C.gloeosporioides sensu lato comprises 33 phylogenetic species and C.gloeosporioides sensu stricto is not a common pathogen of tropical fruits.In this study,we investigated the phylogenetic relationships of 85 Colletotrichum isolates associated with select tropical fruits and flowering plants from India.In the ApMat marker analysis,the 85 isolates clustered with 7 known Colletotrichum species(C.aotearoa,C.dianesei,C.endomangiferae,C.musae,C.siamense,C.theobromicola,Glomerella cingulata f.sp.camelliae)and six novel lineages.One of the novel lineages is described and illustrated in this paper as Colletotrichum communis sp.nov.,while new-host pathogen associations for C.aotearoa,C.endomangiferae,C.dianesei and C.theobromicola are reported from India.Out of the 85 isolates analysed in this paper,73 isolates clustered within the C.siamense species complex,indicating that C.siamense species complex,not C.gloeosporioides sensu stricto,is common on tropical fruits.In comparison with act,cal,gapdh,ITS and tub2 gene markers,we recommend the use of the ApMat marker for accurate identification of cryptic species within the C.siamense species complex.We believe that the ApMat marker,in combination with one or two similar‘phylogenetically superior’gene markers,is a better candidate for specieslevel classification of fungi that were traditionally identified as‘Colletotrichum gloeosporioides’.展开更多
文摘目的:探讨Ig样结构域2黏附分子(adhesion molecule with Ig like domain 2,AMIGO2)在鼻咽癌(nasopharyngeal carcinoma,NPC)细胞增殖中的作用及其机制。方法:选用2017年9月至11月福建省肿瘤医院收集的10例NPC组织和10例正常鼻咽黏膜上皮组织标本,以及NPC细胞系CNE-1、CNE-2、SUNE-1、6-10B、C666-1和人永生化鼻咽黏膜上皮细胞株NP69,用qPCR法检测NPC组织和细胞中AMIGO2 mRNA的表达。构建慢病毒载体干扰AMIGO2表达,用qPCR法验证其干扰效率;用CCK-8法、克隆形成及流式细胞术检测干扰AMIGO2表达对NPC细胞增殖、克隆形成和凋亡的影响,用Western blotting检测干扰AMIGO2表达对NPC细胞增殖及PI3K/AKT/mTOR信号通路相关标志蛋白表达的影响。结果:AMIGO2在NPC组织和CNE-2和SUNE-1细胞中高表达(均P<0.01)。慢病毒AMIGO2感染后,CNE-2和SUNE-1细胞的AMIGO2干扰效率均达50%以上。干扰AMIGO2表达,显著降低CNE-2和SUNE-1细胞增殖及克隆形成能力(均P<0.01)、明显提高细胞的凋亡率(均P<0.01);降低SUNE-1细胞中PI3K、AKT和mTOR磷酸化蛋白的表达水平(均P<0.01)、下调survivin和PCNA蛋白的表达水平(均P<0.01)。结论:AMIGO2通过激活PI3K/AKT/mTOR信号通路促进NPC细胞增殖并抑制其凋亡,提示AMIGO2可能是NPC治疗的潜在靶点。
基金financed by Fundacao de Amparoà Ciência e Tecnologia do Estado de Pernambuc(FACEPE)Coordenacao de Aperfeicoamento de Pessoal de Nível Superior(CAPES)+1 种基金the Conselho Nacional de Desenvolvimento Científico e Tecnológico(CNPq)research fellowshipNRCT of Thailand-Colletotrichum 54201020003 for support.
文摘Endophytic species of Colletotrichum associated with Mangifera indica(mango)are poorly understood.In this study,Colletotrichum species were isolated from mango in Pernambuco State,Brazil.There were significant differences in isolation frequencies of Colletotrichum species among sites and plant tissues.Mature leaf blades were colonized by most Colletotrichum isolates at the majority of sites.Partial sequences of glyceraldehyde-3-phosphate dehydrogenase(GAPDH)of 97 Colletotrichum isolates were amplified as an initial measure of genetic diversity.Phylogenetic analysis with a subset of 22 isolates were performed based on a multilocus dataset(ACT,TUB2,CAL,CHS-1,GAPDH,ITS)followed by Apn2/MAT IGS sequence-analysis for isolates within the C.gloeosporioides species complex.Molecular analysis associated with phenotypic characteristics revealed six previously described species[C.asianum,C.cliviae,C.dianesei(syn.C.melanocaulon),C.fructicola,C.karstii and C.tropicale]and one new species.This new species is introduced as C.endomangiferae.All species isolated were pathogenic on mango fruits but varied in their virulence.There was no distribution pattern of species among sites and plant tissues,although C.asianum was the most prevalent species at all sites and in all plant tissues studied.Five previously reported Colletotrichum species causing anthracnose in mango fruits in northeastern Brazil were also recovered as endophytes.
基金The authors would like to thank CSIR-Institute of Microbial Technology,Chandigarh for the financial support,Dr.D.Ananthapadmanaban for his help in the microscopy and Mr.Deepak Bhatt for DNA sequencing assistance.Drs.Kevin D.Hyde,Lei Cai and Bevan Weir are thanked for the inspiration and useful discussions on Colletotrichum taxonomy.This work was supported by IMTECHOLP0071 project and CSIR-SRF fellowship awarded to GS.This is NIO contribution no.7636 and IMTECH communication no.IMT2014/21.
文摘Colletotrichum gloeosporioides sensu lato has been associated with anthracnose in diverse commercial crops.It is now established that C.gloeosporioides sensu lato comprises 33 phylogenetic species and C.gloeosporioides sensu stricto is not a common pathogen of tropical fruits.In this study,we investigated the phylogenetic relationships of 85 Colletotrichum isolates associated with select tropical fruits and flowering plants from India.In the ApMat marker analysis,the 85 isolates clustered with 7 known Colletotrichum species(C.aotearoa,C.dianesei,C.endomangiferae,C.musae,C.siamense,C.theobromicola,Glomerella cingulata f.sp.camelliae)and six novel lineages.One of the novel lineages is described and illustrated in this paper as Colletotrichum communis sp.nov.,while new-host pathogen associations for C.aotearoa,C.endomangiferae,C.dianesei and C.theobromicola are reported from India.Out of the 85 isolates analysed in this paper,73 isolates clustered within the C.siamense species complex,indicating that C.siamense species complex,not C.gloeosporioides sensu stricto,is common on tropical fruits.In comparison with act,cal,gapdh,ITS and tub2 gene markers,we recommend the use of the ApMat marker for accurate identification of cryptic species within the C.siamense species complex.We believe that the ApMat marker,in combination with one or two similar‘phylogenetically superior’gene markers,is a better candidate for specieslevel classification of fungi that were traditionally identified as‘Colletotrichum gloeosporioides’.
