Abstract
Background: Thymosin β10 is a cytoplasm G-actin sequestering protein. However, its biological functions are largely unknown. In the present study, we evaluated the direct effects of exogenous thymosin β10 on angiogenic potential including endothelial cell migration and capillary-like tube formation. Methods: Human coronary artery endothelial cells (HCAECs) were treated with increasing concentrations of thymosin β10 (25-100 ng/ml) in the upper chamber of a modified Boyden chamber. The cells migrated to the lower chamber were stained with Calcein-AM and assessed by a fluorescence reader. The effect of thymosin β10 on HCAEC capillary-like tube formation was observed on matrigel with basic fibroblast growth factor (bFGF, 20 ng/ml) as a positive control. The tube length was quantitated by Image-Pro plus software and was calculated against untreated groups. Expression of several angiogenic factors including VEGF, VEGFR-1 and integrin-αV in HCAECs before and after thymosin β10 treatment was assessed by real time PCR. Results: Thymosin β10 significantly inhibited cell migration in a dose- and time-dependent manner. Cells treated with thymosin β10 at 100 ng/ml significantly reduced endothelial migration by 36% as compared to controls (P<0.01). In addition, thymosin β10 (100 ng/ml) significantly inhibited the capillary-like tube-formation of HCAECs on matrigel, showing a 21% reduction as compared to controls (P<0.01). Furthermore, real time PCR analysis showed that thymosin β10 significantly decreased mRNA levels of VEGF, VEGFR-1 and integrin αV by 64%, 69% and 67%, respectively, in HCAECs as compared to controls (P<0.05). Conclusions: Thymosin β10 exerts direct inhibitive effects on cell migration and capillary-like tube formation in HCAECs. These effects may be mediated through a decreased expression of angiogenic factors including VEGF, VEGFR-1 and integrin αV. This study suggests a novel function of thymosin β10, which may have potential application in the treatment of diseases with excessive angiogenesis.
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