目的克隆细粒棘球绦虫发育相关基因Tetraspain 2-TSP2(TSP2),并对其进行生物学分析。方法通过在线检索WormBas ParaSite网站中Eg细粒棘球绦虫基因组数据库,获得TSP1的cDNA序列并设计特异性引物。提取细粒棘球绦虫原头蚴总RNA,以此模板,R...目的克隆细粒棘球绦虫发育相关基因Tetraspain 2-TSP2(TSP2),并对其进行生物学分析。方法通过在线检索WormBas ParaSite网站中Eg细粒棘球绦虫基因组数据库,获得TSP1的cDNA序列并设计特异性引物。提取细粒棘球绦虫原头蚴总RNA,以此模板,RT-PCR扩增TSP2基因,将PCR产物克隆到pMD19-T载体后进行测序,并进行生物信息学分析。通过SYBR Green I qRT-PCR方法分析TSP2基因在原头蚴、包囊壁以及成虫中mRNA的相对表达量。结果克隆的TSP2基因序列与WormBas ParaSite中已登录的细粒棘球绦虫跨膜蛋白EGR05083.1同源性为100%。TSP2基因cDNA全长621个核苷酸,编码206个氨基酸的TSP2蛋白,理论等电点为7.33,不稳定指数为47.50,为不稳定蛋白。脂肪系数为109.71,亲水性疏水性平均值为0.987,为疏水性可溶性蛋白。TSP2编码的蛋白含有4个跨膜区,4个优势B抗原表位,属于跨膜蛋白家族,该蛋白具有3个开放阅读框。qRT-PCR显示,TSP2基因在细粒棘球绦虫原头蚴、包囊壁及成虫中均有转录,转录水平差异无统计学意义(P>0.05)。结论细粒棘球绦虫TSP2基因其编码蛋白含有优势B抗原表位,为包虫病重组免疫诊断抗原和亚单位疫苗的研发奠定了基础。展开更多
Background:Gliomas,especially high-grade gliomas,are highly malignant with a poor prognosis.Although existing treatments have improved the survival rate of patients with glioma,the recurrence and mortality rates are s...Background:Gliomas,especially high-grade gliomas,are highly malignant with a poor prognosis.Although existing treatments have improved the survival rate of patients with glioma,the recurrence and mortality rates are still not ideal.The molecular mechanisms involved in the occurrence and development of glioma are still poorly understood.We previously reported that thrombospondin-2(TSP2)expression was increased in tumor specimens from rat models,promoting excitatory synapse formation.However,little is known about the effect of TSP2 on the biological characteristics of glioma.Methods:Glioma and cerebral cortex tissues were collected from 33 patients,and the expression of TSP2 in them was analyzed.Next,the proliferation and migration of TSP2 on glioma cells were analyzed in vitro.At last,a glioma transplantation model was constructed to explore the growth of TSP2 on glioma in vivo.Results:The expression of TSP2 in surgical glioma specimens was increased compared to that in the normal cortex.Interestingly,the TSP2 protein level was higher in high-grade glioma(HGG,World Health Organization(WHO)grades 3-4)than in low-grade glioma(LGG,WHO grades 1-2)tissues.Exogenous addition of the TSP2 protein at an appropriate concentration promoted the migration of glioma cells but did not significantly affect their proliferation.Surprisingly,overexpression of TSP2 promoted both the migration and proliferation of cultured glioma cells.Moreover,in vivo experimental data implied that overexpression of TSP2 in C6 cells promoted the malignant growth of gliomas,while knockout of TSP2 slowed glioma growth.Conclusions:TSP2 promotes the migration and proliferation of glioma cells,which may provide new ideas for blocking glioma progression.展开更多
文摘目的克隆细粒棘球绦虫发育相关基因Tetraspain 2-TSP2(TSP2),并对其进行生物学分析。方法通过在线检索WormBas ParaSite网站中Eg细粒棘球绦虫基因组数据库,获得TSP1的cDNA序列并设计特异性引物。提取细粒棘球绦虫原头蚴总RNA,以此模板,RT-PCR扩增TSP2基因,将PCR产物克隆到pMD19-T载体后进行测序,并进行生物信息学分析。通过SYBR Green I qRT-PCR方法分析TSP2基因在原头蚴、包囊壁以及成虫中mRNA的相对表达量。结果克隆的TSP2基因序列与WormBas ParaSite中已登录的细粒棘球绦虫跨膜蛋白EGR05083.1同源性为100%。TSP2基因cDNA全长621个核苷酸,编码206个氨基酸的TSP2蛋白,理论等电点为7.33,不稳定指数为47.50,为不稳定蛋白。脂肪系数为109.71,亲水性疏水性平均值为0.987,为疏水性可溶性蛋白。TSP2编码的蛋白含有4个跨膜区,4个优势B抗原表位,属于跨膜蛋白家族,该蛋白具有3个开放阅读框。qRT-PCR显示,TSP2基因在细粒棘球绦虫原头蚴、包囊壁及成虫中均有转录,转录水平差异无统计学意义(P>0.05)。结论细粒棘球绦虫TSP2基因其编码蛋白含有优势B抗原表位,为包虫病重组免疫诊断抗原和亚单位疫苗的研发奠定了基础。
基金supported by funds from Scientific and Technological Innovation Talents Project of Sichuan Provincial Science and Technology Department(No.2022JDRC0041)the National Natural Science Foundation of China(No.81772686)+1 种基金Medical Innovation Project(No.21WQ040)Joint Research Project of the General Hospital of Western Theater Command of PLA(No.2019LH01)
文摘Background:Gliomas,especially high-grade gliomas,are highly malignant with a poor prognosis.Although existing treatments have improved the survival rate of patients with glioma,the recurrence and mortality rates are still not ideal.The molecular mechanisms involved in the occurrence and development of glioma are still poorly understood.We previously reported that thrombospondin-2(TSP2)expression was increased in tumor specimens from rat models,promoting excitatory synapse formation.However,little is known about the effect of TSP2 on the biological characteristics of glioma.Methods:Glioma and cerebral cortex tissues were collected from 33 patients,and the expression of TSP2 in them was analyzed.Next,the proliferation and migration of TSP2 on glioma cells were analyzed in vitro.At last,a glioma transplantation model was constructed to explore the growth of TSP2 on glioma in vivo.Results:The expression of TSP2 in surgical glioma specimens was increased compared to that in the normal cortex.Interestingly,the TSP2 protein level was higher in high-grade glioma(HGG,World Health Organization(WHO)grades 3-4)than in low-grade glioma(LGG,WHO grades 1-2)tissues.Exogenous addition of the TSP2 protein at an appropriate concentration promoted the migration of glioma cells but did not significantly affect their proliferation.Surprisingly,overexpression of TSP2 promoted both the migration and proliferation of cultured glioma cells.Moreover,in vivo experimental data implied that overexpression of TSP2 in C6 cells promoted the malignant growth of gliomas,while knockout of TSP2 slowed glioma growth.Conclusions:TSP2 promotes the migration and proliferation of glioma cells,which may provide new ideas for blocking glioma progression.
