The 15(S)-hydroxyeicosatetraenoic acid-induced angiogenesis requires Janus kinase 2-signal transducer and activator of transcription-5B-dependent expression of interleukin-8.
To understand the molecular basis underlying 15(S)-hydroxyeicosatetraenoic acid (15(S)-HETE)-induced angiogenesis, we have studied the role of the Janus kinase-signal transducer and activator of transcription (Jak-STAT) signaling. The 15(S)-HETE stimulated tyrosine phosphorylation of Jak2 in a time-dependent manner in human retinal microvascular endothelial cells (HRMVECs). Inhibition of Jak2 activation via adenovirus-mediated expression of its dominant-negative mutant attenuated 15(S)-HETE-induced HRMVEC migration and tube formation and Matrigel plug angiogenesis. Similarly, 15(S)-HETE activated tyrosine phosphorylation of STAT-5B in a time-dependent manner. Dominant-negative mutant-mediated interference of STAT-5B activation suppressed 15(S)-HETE-induced HRMVEC migration and tube formation and Matrigel plug angiogenesis. The 15(S)-HETE induced interleukin-8 (IL-8) expression in Jak2-STAT-5B-dependent manner in HRMVECs. In addition, neutralizing anti-IL-8 antibodies reduced 15(S)-HETE-induced HRMVEC migration and tube formation and Matrigel plug angiogenesis. Cloning and Transfac analysis of IL-8 promoter revealed the presence of 1 putative STAT-binding sequence at -476 nt, and electrophoretic mobility shift assay and chromatin immunoprecipitation analysis showed the binding of STAT-5B to this site in response to 15(S)-HETE. Mutational analysis showed that STAT binding site is essential for 15(S)-HETE-induced IL-8 promoter activity. Together, these observations suggest that 15(S)-HETE-induced angiogenesis requires Jak2-STAT-5B-dependent expression of IL-8.
Department of Physiology, University of Tennessee Health Science Center, Memphis, TN 38163, USA.
Sergey Y
SY
Cheranov
Dong
D
Wang
Venkatesh
V
Kundumani-Sridharan
Manjula
M
Karpurapu
Qiuhua
Q
Zhang
Koteswara R
KR
Chava
Gadiparthi N
GN
Rao
D001483:Base Sequence; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D005786:Gene Expression Regulation / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000494:pharmacology*; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism*; D053614:Janus Kinase 2 / Q000378:metabolism*; D008969:Molecular Sequence Data; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D050799:STAT5 Transcription Factor / Q000235:genetics / Q000378:metabolism*
D006893:Hydroxyeicosatetraenoic Acids; D016209:Interleukin-8; D050799:STAT5 Transcription Factor; C494095:STAT5B protein, human; C025984:15-hydroxy-5,8,11,13-eicosatetraenoic acid; D053614:Janus Kinase 2