TY - JOUR
T1 - Involvement of BIG1 and BIG2 in regulating VEGF expression and angiogenesis
AU - Lu, Fu I.
AU - Wang, Yi Ting
AU - Wang, Yi Shan
AU - Wu, Chang Yi
AU - Li, Chun Chun
N1 - Funding Information:
The authors acknowledge the help of Drs. Wen-Tai Chiu (National Cheng Kung University) and Won-Jing Wang (National Yang Ming University, Taipei, Taiwan) for commenting on this work. The authors thank Drs. Chia-Jung Yu (Chang Gung University, Taoyuan City, Taiwan), Jean-Chen Kuo (National Yang Ming University), and Ya-Wen Liu (National Taiwan University, Taipei, Taiwan) for critical review of this manuscript. The authors also thank Hsiao-Han Huang and Yu-Fang Wang (both from National Cheng Kung University) for technique support. The authors also thank the Taiwan Zebrafish Core Facility at Academia Sinica (TZCAS) and National Health Research Institute (TZeTH), which are supported by Ministry of Science and Technology (MOST105-2321-B-001-029 and MOST104-2321-B-001-045) for providing wild-type zebrafish and plasmids. The authors are also grateful for the support from the Laboratory Animal Center, Medical College, National Cheng Kung University. This work was supported by grants from Ministry of Science and Technology in Taiwan (MOST) awarded to F.-I.L. (MOST 104-2311-B-006-005-MY3; MOST 107-2311-B.006-001) and to C.-C.L. (MOST 104-2320-B-006-032-MY3; MOST 107-2320-B-006-041). This work was, in part, financially supported by the Integrative Evolutionary Galliforms Genomics Research (iEGG) and Animal Biotechnology Center from The Feature Area Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (MOE) in Taiwan (MOE-107-S-0023-F). The authors declare no conflicts of interest.
Publisher Copyright:
© FASEB
PY - 2019/9/1
Y1 - 2019/9/1
N2 - VEGF stimulates the formation of new blood vessels by inducing endothelial cell (EC) proliferation and migration. Brefeldin A (BFA)-inhibited guanine nucleotide–exchange protein (BIG)1 and 2 accelerate the replacement of bound GDP with GTP to activate ADP-ribosylation factor (Arf)1, which regulates vesicular transport between the Golgi and plasma membrane. Although it has been reported that treating cells with BFA interferes with Arf1 activation to inhibit VEGF secretion, the role of BIG1 and BIG2 in VEGF trafficking and expression, EC migration and proliferation, and vascular development remains unknown. Here, we found that inactivation of Arf1 reduced VEGF secretion but did not affect the levels of VEGF protein. Interestingly, however, BIG1 and BIG2 knockdown significantly decreased the levels of VEGF mRNA and protein in glioblastoma U251 cells and HUVECs. Furthermore, depletion of BIG1 and BIG2 inhibited HUVEC angiogenesis by diminishing cell migration. Angioblast migration and intersegmental vessel sprouting were also impaired when the BIG2 homolog, Arf guanine nucleotide exchange factor (arfgef)2, was knocked down in zebrafish with endothelial expression of green fluorescent protein (GFP). Depletion of arfgef2 by clustered regularly interspaced short palindromic repeat (CRISPR)/ CRISPR-associated protein 9 (Cas9) also caused defects in vascular development of zebrafish embryos. Taken together, these data reveal that BIG1 and BIG2 participate in endothelial cell angiogenesis.—Lu, F.-L, Wang, Y.-T., Wang, Y.-S., Wu, C.-Y., Li, C.-C. Involvement of BIG1 and BIG2 in regulating VEGF expression and angiogenesis. FASEB J. 33, 9959–9973 (2019). www.fasebj.org.
AB - VEGF stimulates the formation of new blood vessels by inducing endothelial cell (EC) proliferation and migration. Brefeldin A (BFA)-inhibited guanine nucleotide–exchange protein (BIG)1 and 2 accelerate the replacement of bound GDP with GTP to activate ADP-ribosylation factor (Arf)1, which regulates vesicular transport between the Golgi and plasma membrane. Although it has been reported that treating cells with BFA interferes with Arf1 activation to inhibit VEGF secretion, the role of BIG1 and BIG2 in VEGF trafficking and expression, EC migration and proliferation, and vascular development remains unknown. Here, we found that inactivation of Arf1 reduced VEGF secretion but did not affect the levels of VEGF protein. Interestingly, however, BIG1 and BIG2 knockdown significantly decreased the levels of VEGF mRNA and protein in glioblastoma U251 cells and HUVECs. Furthermore, depletion of BIG1 and BIG2 inhibited HUVEC angiogenesis by diminishing cell migration. Angioblast migration and intersegmental vessel sprouting were also impaired when the BIG2 homolog, Arf guanine nucleotide exchange factor (arfgef)2, was knocked down in zebrafish with endothelial expression of green fluorescent protein (GFP). Depletion of arfgef2 by clustered regularly interspaced short palindromic repeat (CRISPR)/ CRISPR-associated protein 9 (Cas9) also caused defects in vascular development of zebrafish embryos. Taken together, these data reveal that BIG1 and BIG2 participate in endothelial cell angiogenesis.—Lu, F.-L, Wang, Y.-T., Wang, Y.-S., Wu, C.-Y., Li, C.-C. Involvement of BIG1 and BIG2 in regulating VEGF expression and angiogenesis. FASEB J. 33, 9959–9973 (2019). www.fasebj.org.
UR - http://www.scopus.com/inward/record.url?scp=85071783926&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85071783926&partnerID=8YFLogxK
U2 - 10.1096/fj.201900342RR
DO - 10.1096/fj.201900342RR
M3 - Article
C2 - 31199673
AN - SCOPUS:85071783926
VL - 33
SP - 9959
EP - 9973
JO - FASEB Journal
JF - FASEB Journal
SN - 0892-6638
IS - 9
ER -