Eye Institute, Affiliated Hospital of Nantong University, Nantong, China.
Curr Eye Res. 2022 Oct;47(10):1381-1388. doi: 10.1080/02713683.2022.2108455. Epub 2022 Aug 3.
To explore the effect of doxycycline on vasculogenic mimicry (VM) formation and the potential mechanism in human pterygium fibroblasts in order to find novel targets for pterygium therapy.
First, we demonstrate the existence of VM in 73 pterygium specimens by CD31 and periodic acid Schiff (PAS) dual staining. Then we used cell counting kit-8, clone formation assay and flow cytometry to prove the inhibitory effect of doxycycline on cell proliferation and apoptosis. The VM formation was evaluated through wound healing assay, cell transwell assay and three-dimensional cell culture combined with PAS staining. Finally, we used Western blot to testify the correlation of the VM and the factors in protein level preliminarily.
Our results showed that VM existed in human pterygium specimens exactly. Otherwise, in human pterygium fibroblasts, doxycycline induced a dose-dependent inhibitory effect on cell proliferation and apoptosis induction. Besides, doxycycline significantly suppressed vasculogenic mimicry tube formation, cell migration and invasion. Furthermore, doxycycline impaired the expression of MMP-9, MMP-2 and VEGF which may related to pterygium VM formation.
Doxycycline decelerated pterygium progression might be through inhibiting VM formation according to the downregulation of MMP-9, MMP-2 and VEGF, which may provide the basis of further studies involving doxycycline for pterygium treatment.
探讨强力霉素对人翼状胬肉成纤维细胞血管生成拟态(VM)形成的影响及其潜在机制,以期为翼状胬肉的治疗寻找新的靶点。
首先,我们通过 CD31 和过碘酸希夫(PAS)双重染色证实了 73 例翼状胬肉标本中 VM 的存在。然后,我们使用细胞计数试剂盒-8、克隆形成实验和流式细胞术证明强力霉素对细胞增殖和凋亡的抑制作用。通过划痕愈合实验、细胞 Transwell 实验和与 PAS 染色相结合的三维细胞培养来评估 VM 的形成。最后,我们使用 Western blot 初步验证了 VM 与蛋白水平相关因子的相关性。
我们的结果表明,VM 确实存在于人翼状胬肉标本中。此外,在人翼状胬肉成纤维细胞中,强力霉素呈剂量依赖性地抑制细胞增殖并诱导细胞凋亡。此外,强力霉素显著抑制血管生成拟态管形成、细胞迁移和侵袭。此外,强力霉素削弱了 MMP-9、MMP-2 和 VEGF 的表达,这可能与翼状胬肉 VM 的形成有关。
强力霉素通过下调 MMP-9、MMP-2 和 VEGF 来减缓翼状胬肉的进展,这可能为进一步研究强力霉素治疗翼状胬肉提供依据。