摘要
目的 TEAD1转录因子1 (TEAD1)在前脂肪细胞中表达,但是功能还不清楚。本研究旨在探讨TEAD1的两个转录本对永生化鸡前脂肪细胞系(immortalized chicken preadipocyte 1,ICP1)细胞增殖、迁移、凋亡和分化的影响。方法 克隆TEAD1基因的全长序列,对获得的两个转录本进行生物信息学分析。利用间接免疫荧光分析TEAD1转录本的亚细胞定位。通过RT-qPCR、CCK-8和EdU等方法,检测过表达TEAD1转录本对ICP1细胞增殖的影响。利用划痕实验检测TEAD1转录本对ICP1细胞迁移的影响。利用细胞凋亡-Hoechst染色和RT-qPCR,分析过表达TEAD1转录本对ICP1细胞凋亡的影响。通过RT-qPCR检测TEAD1转录本在不同组织、不同细胞系和ICP1细胞分化过程中的表达。利用油红O染色、BODIPY染色、RT-qPCR、Western blot和双荧光素酶报告基因技术,分析过表达TEAD1转录本对ICP1细胞脂滴积累及成脂相关基因转录的影响。最后,测定过表达TEAD1转录本的ICP1细胞中甘油三酯(TG)含量。结果 克隆TEAD1全长编码区,鉴定出两个TEAD1转录本。TEAD1-V1主要定位于细胞核,TEAD1-V2定位于细胞质与细胞核。过表达TEAD1-V1和TEAD1-V2均抑制ICP1细胞增殖。过表达TEAD1-V1促进ICP1细胞迁移,而过表达TEAD1-V2对ICP1细胞迁移没有影响。且过表达TEAD1-V1和TEAD1-V2均促进ICP1细胞凋亡。两个转录本在不同组织和细胞系中的表达模式相似,在前脂肪细胞分化过程中的表达先下降后上升。过表达TEAD1-V1能显著减少ICP1细胞中脂滴的积累(P<0.05),抑制C/EBPα表达;而过表达TEAD1-V2对脂滴积累和成脂相关基因的蛋白质表达水平没有明显作用(P>0.05)。过表达TEAD1-V1能显著降低ICP1细胞中甘油三脂的含量(P<0.05),而过表达TEAD1-V2对ICP1细胞中甘油三酯含量没有影响(P>0.05)。结论 本研究首次克隆并鉴定了鸡TEAD1基因的两个转录本。过表达转录本TEAD1-V1和TEAD1-V2抑制鸡前脂肪细胞增殖并且促进鸡�
Objective Although expression of the TEAD1 protein in preadipocytes has been established,its function remains unclear.In this study,we sought to detect transcripts of TEAD1 in chicken and to examine the effects of this protein on the proliferation,migration,apoptosis,and differentiation of immortalized chicken preadipocyte cell lines(ICP1).Methods The full-length sequence of the TEAD1 gene was cloned and the two transcripts were subjected to bioinformatics analysis.The subcellsular localization of TEAD1 transcripts was determined based on indirect immunofluorescence.The effects of TEAD1 transcripts overexpression on the proliferation of ICP1 cells were examined by RT-qPCR,CCK-8,and EdU assays;the effects of TEAD1 transcripts on ICP1 cells migration were examined based on the scratch test;and the effects of TEAD1 transcripts overexpression on ICP1 cells apoptosis were analyzed using apoptosis-Hoechst staining and RT-qPCR.The expression of TEAD1 transcripts in different tissues,cells lines,and ICP1 at different periods of differentiation was analyzed by RT-qPCR.The effects of TEAD1 transcripts overexpression on lipid droplet accumulation and adipogenic-related gene expression in ICP1 cells were analyzed based on Oil Red O and BODIPY staining,RT-qPCR,Western blot,and dual-luciferase reporter gene assays.Finally,the content of triglyceride(TG)was measured in TEAD1 overexpressed ICP1 cells.Results The full-length TEAD1 was cloned and two TEAD1 transcripts were identified.The TEAD1-V1 protein was found to be localized primarily in the cell nucleus,whereas the TEAD1-V2 protein is localized in the cell cytoplasm and nucleus.The overexpression of both TEAD1-V1 and TEAD1-V2 significantly inhibited the proliferation of ICP1 cells.Whereas the overexpression of TEAD1-V1 promoted ICP1 cell migration,the overexpression of TEAD1-V2 had no significant effects on ICP1 migration;the overexpression of both TEAD1-V1 and TEAD1-V2 significantly promoted the apoptosis of ICP1 cells.We found that the different transcripts of TEAD1 have s
作者
彭敏
徐虎
贾子秋
杨清竹
潘林
王维禹
孔令喆
孙婴宁
PENG Min;XU Hu;JIA Zi-Qiu;YANG Qing-Zhu;PAN Lin;WANG Wei-Yu;KONG Ling-Zhe;SUN Ying-Ning(Department of Biotechnology,College of Life Science and Agriculture and Forestry,Qiqihar University,Qiqihar 161006,China)
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2024年第1期215-229,共15页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金(31402061)
黑龙江省自然科学基金(YQ2019C025)
黑龙江省省属高等学校基本科研业务费(145109211)资助项目。
