Elevated Expression of miR302-367 in Endothelial Cells Inhibits Developmental Angiogenesis via CDC42/CCND1 Mediated Signaling Pathways

被引：16

作者：

Pi, Jingjiang ^{[1
]}

Liu, Jie ^{[1
]}

Zhuang, Tao ^{[1
]}

Zhang, Lin ^{[1
]}

Sun, Huimin ^{[1
]}

Chen, Xiaoli ^{[1
]}

Zhao, Qian ^{[1
]}

Kuang, Yashu ^{[1
]}

Peng, Sheng ^{[1
]}

Zhou, Xiaohui ^{[1
]}

Yu, Zuoren ^{[1
]}

Tao, Ting ^{[2
]}

Tomlinson, Brian ^{[3
]}

Chan, Paul ^{[4
]}

Tian, Ying ^{[5
]}

Fan, Huimin ^{[1
]}

Liu, Zhongmin ^{[1
]}

Zheng, Xiangjian ^{[6
]}

Morrisey, Edward ^{[7
,8
,9
,10
]}

Zhang, Yuzhen ^{[1
]}

机构：

[1] Tongji Univ, Shanghai East Hosp, Res Ctr Translat Med, Key Lab Arrhythmias,Minist Educ China,Sch Med, 150 Jimo Rd, Shanghai 200120, Peoples R China

[2] Jiaotong Univ, Sch Med, Ruijin Hosp, Dept Geriatr, Shanghai 200025, Peoples R China

[3] Chinese Univ Hong Kong, Dept Med & Therapeut, Hong Kong, Hong Kong, Peoples R China

[4] Taipei Med Univ, Wan Fang Hosp, Div Cardiol, Dept Internal Med, Taipei, Taiwan

[5] Temple Univ, Dept Pharmacol, Ctr Translat Med, Sch Med, Philadelphia, PA 19140 USA

[6] Centenary Inst, Lab Cardiovasc Signaling, Camperdown, NSW 2050, Australia

[7] Univ Penn, Dept Cell & Dev Biol, Philadelphia, PA 19104 USA

[8] Univ Penn, Dept Med, Philadelphia, PA 19104 USA

[9] Univ Penn, Penn Cardiovasc Inst, Philadelphia, PA 19104 USA

[10] Univ Penn, Penn Inst Regenerat Med, Philadelphia, PA 19104 USA

来源：

THERANOSTICS | 2018年 / 8卷 / 06期

基金：

中国国家自然科学基金;

关键词：

miR302-367; endothelial cells; developmental angiogenesis; cdc42; actin remodeling; cell cycle; ARP2/3; COMPLEX; RHO GTPASES; STEM-CELLS; PAK1; MIGRATION; MECHANISMS; KINASE; P21-ACTIVATED-KINASE-1; CYTOSKELETON; DYNAMICS;

D O I：

10.7150/thno.21986

中图分类号：

R-3 [医学研究方法]; R3 [基础医学];

学科分类号：

1001 ;

摘要：

Rationale: Angiogenesis is critical for embryonic development and microRNAs fine-tune this process, but the underlying mechanisms remain incompletely understood. Methods: Endothelial cell (EC) specific miR302-367 line was used as gain-of-function and anti-miRs as loss-of-function models to investigate the effects of miR302-367 on developmental angiogenesis with embryonic hindbrain vasculature as an in vivo model and fibrin gel beads and tube formation assay as in vitro models. Cell migration was evaluated by Boyden chamber and scratch wound healing assay and cell proliferation by cell count, MTT assay, Ki67 immunostaining and PI cell cycle analysis. RNA high-throughput sequencing identified miR-target genes confirmed by chromatin immunoprecipitation and 3'-UTR luciferase reporter assay, and finally target site blocker determined the pathway contributing significantly to the phenotype observed upon microRNA expression. Results: Elevated EC miR302-367 expression reduced developmental angiogenesis, whereas it was enhanced by inhibition of miR302-367, possibly due to the intrinsic inhibitory effects on EC migration and proliferation. We identified Cdc42 as a direct target gene and elevated EC miR302-367 decreased total and active Cdc42, and further inhibited F-actin formation via the WASP and Klf2/Grb2/Pak1/LIM-kinase/Cofilin pathways. MiR302-367-mediated-Klf2 regulation of Grb2 for fine-tuning Pak1 activation contributing to the inhibited F-actin formation, and then the attenuation of EC migration. Moreover, miR302-367 directly down-regulated EC Ccnd1 and impaired cell proliferation via the Rb/E2F pathway. Conclusion: miR302-367 regulation of endothelial Cdc42 and Ccnd1 signal pathways for EC migration and proliferation advances our understanding of developmental angiogenesis, and meanwhile provides a rationale for future interventions of pathological angiogenesis that shares many common features of physiological angiogenesis.

引用

页码：1511 / 1526

页数：16

共 40 条

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