基础医学与临床 ›› 2021, Vol. 41 ›› Issue (6): 859-864.

• 研究论文 • 上一篇    下一篇

DFG-STYK1/NOK对小鼠胚胎成纤维细胞系NIH3T3增殖以及细胞周期G1/S的影响

陈建苗*   

  1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 病原生物学系, 北京 100005
  • 收稿日期:2020-12-25 修回日期:2021-04-16 出版日期:2021-06-05 发布日期:2021-05-31
  • 通讯作者: *cjm1906101412@163.com
  • 基金资助:
    中国医学科学院创新工程(2017-12M-3-007)

Influence of DFG-STYK1/NOK on cell proliferation and G1/S distribution in mouse fibroblast cell line NIH3T3

CHEN Jian-miao*   

  1. Department of Etiology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005,China
  • Received:2020-12-25 Revised:2021-04-16 Online:2021-06-05 Published:2021-05-31
  • Contact: *cjm1906101412@163.com

摘要: 目的 研究新型受体酪氨酸激酶STYK1/NOK加入DFG基序后对小鼠胚胎成纤维细胞系NIH3T3增殖和细胞周期G1/S的影响。方法 SWISS MODEL同源比对STYK1/NOK的二级结构确定DFG加入的位置;用CCK8和MTT法检测瞬时转染pcDNA3.0或pcDNA3.0-DFG-NOK的NIH3T3细胞增殖;流式细胞测量术检测细胞周期;Western blot检测细胞增殖信号通路蛋白Akt、p-Akt、Erk、p-Erk、STAT、p-STAT、JAK3、p-JAK3和周期蛋白cyclin D1的表达。结果 与瞬时转染pcDNA3.0相比,1)瞬时转染pcDNA3.0-DFG-NOK可抑制NIH3T3细胞增殖;2)DFG-NOK阻碍细胞周期由G1期进入S期;3)DFG-NOK抑制Erk、STAT1、STAT3和JAK3的磷酸化和周期蛋白cyclin D1表达。结论 DFG-NOK抑制NIH3T3细胞增殖和细胞周期G1期进入S期。

关键词: DFG-STYK1/NOK, NIH3T3细胞, 细胞增殖, 信号通路, G1/S

Abstract: Objective To investigate the effects of NOK added DFG motif on proliferation and G1/S distribution of NIH3T3 cells. Methods SWISS MODEL homologous modeling was used to confirm the location of DFG addition; we used CCK-8 and MTT methods to measure the cell proliferation of NIH3T3 transiently transfected pcDNA3.0 or pcDNA3.0-DFG-NOK; Flow cytometry was used to measure cell cycle; Western-blot was used to exam the phosphorylation of Akt,Erk,STAT1,STAT3,JAK3 and the expression of cyclin D1. Results Compared with the pcDNA3.0 transient transfection: 1)The proliferation of pcDNA3.0-DFG-NOK over-expression NIH3T3 cells was inhibited. 2)The entrance of G1/S in NIH3T3 cells transiently transfected pcDNA3.0-DFG-NOK was obstructed. 3)Transient transfection of pcDNA3.0-DFG-NOK could inhibit phosphorylation of Erk,STAT1,STAT3,JAK3 and expression of CyclinD1. Conclusions DFG-NOK inhibits proliferation of NIH3T3 cells and obstructs cell cycle G1 phase entering into S phase.

Key words: DFG-STYK1/NOK, NIH3T3 cells, cell proliferation, signaling pathway, G1/S

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