Cathepsin L derived from skeletal muscle cells transfected with bFGF promotes endothelial cell migration

Ji Hyung Chung, Eun Kyoung Im, Taewon Jin, Seung Min Lee, Soo Hyuk Kim, Eun Young Choi, Min-Jeong Shin, Kyung Hye Lee, Yangsoo Jang

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Gene transfer of basic fibroblast growth factor (bFGF) has been shown to induce significant endothelial migration and angiogenesis in ischemic disease models. Here, we investigate what factors are secreted from skeletal muscle cells (SkMCs) transfected with bFGF gene and whether they participate in endothelial cell migration. We constructed replication-defective ad-enovirus vectors containing the human bFGF gene (Ad/bFGF) or a control LacZ gene (Ad/LacZ) and obtained conditioned media, bFGF-CM and LacZ-CM, from SkMCs infected by Ad/bFGF or Ad/LacZ, respectively. Cell migration significantly increased in HUVECs incubated with bFGF-CM compared to cells incubated with LacZ-CM. Interestingly, HUVEC migration in response to bFGF-CM was only partially blocked by the addition of bFGF-neutralizing antibody, suggesting that bFGF-CM contains other factors that stimulate endothelial cell migration. Several proteins, matrix metalloproteinase-1 (MMP-1), plasminogen activator inhibitor-1 (PAI-1), and cathepsin L, increased in bFGF-CM compared to LacZ-CM; based on 1-dimensional gel electrophoresis and mass spectrometry. Their increased mRNA and protein levels were confirmed by RT-PCR and immunoblot analysis. The recombinant human bFGF protein induced MMP-1, PAI-1, and cathepsin L expression in SkMCs. Endothelial cell migration was reduced in groups treated with bFGF-CM containing neutralizing antibodies against MMP-1 or PAI-1. In particular, HUVECs treated with bFGF-CM containing cell-impermeable cathepsin L inhibitor showed the most significant decrease in cell migration. Cathepsin L protein directly promotes endothelial cell migration through the JNK pathway. These results indicate that cathepsin L released from SkMCs transfected with the bFGF gene can promote endothelial cell migration.

Original languageEnglish
Pages (from-to)179-188
Number of pages10
JournalExperimental and Molecular Medicine
Volume43
Issue number4
DOIs
Publication statusPublished - 2011 Apr 1

Fingerprint

Cathepsin L
Endothelial cells
Fibroblast Growth Factor 2
Muscle Cells
Cell Movement
Muscle
Skeletal Muscle
Endothelial Cells
Cells
Matrix Metalloproteinase 1
Plasminogen Activator Inhibitor 1
Genes
Neutralizing Antibodies
Proteins
Gene transfer
Lac Operon
MAP Kinase Signaling System
Conditioned Culture Medium
Electrophoresis

Keywords

  • Cathepsin L
  • Cell migration
  • Endothelium
  • Fibroblast growth factor 2
  • JNK mitogen-activated protein kinases

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Molecular Medicine
  • Clinical Biochemistry

Cite this

Cathepsin L derived from skeletal muscle cells transfected with bFGF promotes endothelial cell migration. / Chung, Ji Hyung; Im, Eun Kyoung; Jin, Taewon; Lee, Seung Min; Kim, Soo Hyuk; Choi, Eun Young; Shin, Min-Jeong; Lee, Kyung Hye; Jang, Yangsoo.

In: Experimental and Molecular Medicine, Vol. 43, No. 4, 01.04.2011, p. 179-188.

Research output: Contribution to journalArticle

Chung, Ji Hyung ; Im, Eun Kyoung ; Jin, Taewon ; Lee, Seung Min ; Kim, Soo Hyuk ; Choi, Eun Young ; Shin, Min-Jeong ; Lee, Kyung Hye ; Jang, Yangsoo. / Cathepsin L derived from skeletal muscle cells transfected with bFGF promotes endothelial cell migration. In: Experimental and Molecular Medicine. 2011 ; Vol. 43, No. 4. pp. 179-188.
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