Ena/VASP样蛋白(Ena/VASP like protein,EVL)是Ena/VASP家族成员之一,它参与肌动蛋白细胞骨架重组,以及细胞迁移、收缩环形成和细胞间附着.EVL在肝癌SMMC-7721细胞中高表达.抑制EVL蛋白表达后,SMMC-7721细胞的增殖与迁移能力降低.为研究...Ena/VASP样蛋白(Ena/VASP like protein,EVL)是Ena/VASP家族成员之一,它参与肌动蛋白细胞骨架重组,以及细胞迁移、收缩环形成和细胞间附着.EVL在肝癌SMMC-7721细胞中高表达.抑制EVL蛋白表达后,SMMC-7721细胞的增殖与迁移能力降低.为研究EVL在肝癌细胞的功能,构建了靶向shRNA干扰表达载体,稳定转染肝癌SMMC-7721细胞.MTT实验和细胞集落形成实验显示,与转染对照比较,沉默EVL蛋白表达可明显抑制SMMC-7721肝癌细胞的增殖、集落形成能力.Transwell实验证明,沉默EVL表达导致SMMC-7721细胞迁移能力降低.进而,流式细胞术揭示,沉默EVL表达的SMMC-7721细胞G0/G1期细胞比例增多.研究结果提示,EVL蛋白可促进肝癌细胞的增殖与迁移;该结果可解释EVL在肝癌细胞中高表达的意义.展开更多
Epiboly,the coordinated spreading of the enveloping layer(EVL),the yolk syncytial layer(YSL)and the deep cells(DCs)toward the vegetal pole,is required for gastrulation.Interference with the EVL leads to early developm...Epiboly,the coordinated spreading of the enveloping layer(EVL),the yolk syncytial layer(YSL)and the deep cells(DCs)toward the vegetal pole,is required for gastrulation.Interference with the EVL leads to early developmental failure.However,the genetic factors underlying EVL differentiation are so far unknown.By in-depth analyzing of single-cell sequencing data from 4 h post fertilization zebrafish embryos,we identified 105 genes(FDR<0.05)which showed highly specific EVL expression,among which the transcription factor cebpb and genes(krt18,cldne,cldnb,krt4,krt5)involved in cell-cell junction formation were exclusively expressed in EVL.To explore the role of cebpb in EVL differentiation,we knocked down its expression by morpholino-labeled antisense RNA.The morphant embryos showed disrupted EVL and developmental termination prior to epiboly.In accordance,expression of the EVL specific cell-adhesion molecule genes were significantly reduced.Thus,cebpb is the earliest EVL marker and essential for EVL differentiation in zebrafish.The elucidation of the specific EVL gene set provides us new insights into the molecular mechanisms underlying EVL differentiation.展开更多
文摘Ena/VASP样蛋白(Ena/VASP like protein,EVL)是Ena/VASP家族成员之一,它参与肌动蛋白细胞骨架重组,以及细胞迁移、收缩环形成和细胞间附着.EVL在肝癌SMMC-7721细胞中高表达.抑制EVL蛋白表达后,SMMC-7721细胞的增殖与迁移能力降低.为研究EVL在肝癌细胞的功能,构建了靶向shRNA干扰表达载体,稳定转染肝癌SMMC-7721细胞.MTT实验和细胞集落形成实验显示,与转染对照比较,沉默EVL蛋白表达可明显抑制SMMC-7721肝癌细胞的增殖、集落形成能力.Transwell实验证明,沉默EVL表达导致SMMC-7721细胞迁移能力降低.进而,流式细胞术揭示,沉默EVL表达的SMMC-7721细胞G0/G1期细胞比例增多.研究结果提示,EVL蛋白可促进肝癌细胞的增殖与迁移;该结果可解释EVL在肝癌细胞中高表达的意义.
基金supported by National Key Research and Development Program of China[2018YFD0900601]L.Chen,the open project fund from Laboratory for Marine Biology and Biotechnology,Qingdao National Laboratory for Marine Science(OF2019NO01)+1 种基金Doctor Fund of Shanghai Ocean University(A2-2006-21-200304)Science and Technology Development Fund of Shanghai Ocean University(A2-2006-20-200201)to W.Li.
文摘Epiboly,the coordinated spreading of the enveloping layer(EVL),the yolk syncytial layer(YSL)and the deep cells(DCs)toward the vegetal pole,is required for gastrulation.Interference with the EVL leads to early developmental failure.However,the genetic factors underlying EVL differentiation are so far unknown.By in-depth analyzing of single-cell sequencing data from 4 h post fertilization zebrafish embryos,we identified 105 genes(FDR<0.05)which showed highly specific EVL expression,among which the transcription factor cebpb and genes(krt18,cldne,cldnb,krt4,krt5)involved in cell-cell junction formation were exclusively expressed in EVL.To explore the role of cebpb in EVL differentiation,we knocked down its expression by morpholino-labeled antisense RNA.The morphant embryos showed disrupted EVL and developmental termination prior to epiboly.In accordance,expression of the EVL specific cell-adhesion molecule genes were significantly reduced.Thus,cebpb is the earliest EVL marker and essential for EVL differentiation in zebrafish.The elucidation of the specific EVL gene set provides us new insights into the molecular mechanisms underlying EVL differentiation.
