Sonic hedgehog protein promotes bone marrow-derived endothelial progenitor cell proliferation, migration and VEGF production via PI 3-kinase/ Akt signaling pathways1

Abstract

Aim: To investigate the effects of Sonic hedgehog (shh) protein on bone marrow‐derived endothelial progenitor cells (BM‐EPC) proliferation, migration and vascular endothelial growth factor (VEGF) production, and the potential signaling pathways involved in these effects. Methods: Bone marrow‐derived Flk‐1+ cells were enriched using the MACS system from adult Kunming mice and then BM‐EPC was cultured in gelatin‐coated culture dishes. The effects of shh N‐terminal pep‐tide on BM‐EPC proliferation were evaluated using the MTT colorimetric assay. Cell migration was assayed using a modified Boy den chamber technique. The production of VEGF was determined by ELISA and immunofluorescence analysis. The potential involvement of PKC and PI3K signaling pathways was explored using selective inhibitor or Western blot. Results: The proliferation, migration and VEGF production in BM‐EPC could be promoted by endogenous shh N‐terminal peptide at concentrations of 0.1 ug/mL to 10 ug/mL, and could be inhibited by anti‐shh antibodies. Shh‐mediated proliferation and migration in BM‐EPC could be partly attenuated by anti‐VEGF. Phospho‐PI3 ‐kinase expression in newly separated BM‐EPC was low, and it increased significantly when exogenous shh N‐terminal peptide was added, but could be attenuated by anti‐human/mouse shh N‐terminal peptide antibody. Moreover, the inhibitor of the PI3‐kinase, but not the inhibitor of the PKC, significantly inhibited the shh‐mediated proliferation, migration and VEGF production. Conclusion: Shh protein can stimulate bone marrow‐derived BM‐EPC proliferation, migration and VEGF production, which may promote neovascularization to ischemic tissues. This results also suggests that the PI3‐kinase/Akt signaling pathways are involved in the angiogenic effects of shh.

Sonic hedgehog mediates a novel pathway of PDGF-BB–dependent vessel maturation

“…21,23,[28][29][30] We demonstrated that recombinant Shh is capable of inducing a fast (15-minute) Akt phosphorylation in SMCs. We showed that Shhinduced Akt phosphorylation is entirely inhibited by the pan PI3K inhibitor Wortmannin, the PI3Kg selective inhibitor AS-252424 ( Figure 3A), and competition with the inactive mutant of PI3Kg 25 (supplemental Figure 7A), demonstrating that Shh-induced Akt phosphorylation is mediated via PI3Kg.…”

“…At the cellular level, recombinant Shh was shown to induce SMC and pericyte proliferation and survival [16][17][18][19] and to contribute to PDGF-BB-induced myofibroblast proliferation. 20 Even though Shh was shown to induce the migration of various cell types such as fibroblasts, 21 ECs, 22 endothelial progenitors, 23 and monocytes, 24 its role and mechanism in SMC migration in vitro and in MC recruitment in vivo have never been investigated.…”

Endothelial Precursor Cell Migration During Vasculogenesis

“…It has been suggested that the phosphatidylinositol-kinase/Akt signaling pathways are involved in such effects of sonic hedgehog. 45 Evidence on the involvement of CXCR4 and the ligand SDF-1 in the regulation of endothelial precursor migration also comes from investigations of CXCR4/SDF-1 signaling in EPCs. It has been shown that CXCR4 is expressed in EPCs and SDF-1 can induce increased migration of EPCs mediated by CXCR4.…”

Role of Hedgehog signaling pathway in proliferation and invasiveness of hepatocellular carcinoma cells

“…Although it has been confirmed that the Hh pathway participates in cell migration and angiogenesis processes and is correlated with the severity of the associated tumor (27)(28)(29), additionally, disruption of Hh pathway has been shown to inhibit metastasis in pancreatic cancer and gastric cancer (29)(30)(31), the relationship between the Hh signaling pathway and aggressiveness of HCC cells remains unclear. It is necessary to investigate whether the signaling influences proliferative and invasive potential of HCC cells.…”