Department of Burns and Reconstructive Surgery, Xiangya Hospital, Central South University, Changsha, Hunan 410008, PR China.
Department of Burns and Reconstructive Surgery, Xiangya Hospital, Central South University, Changsha, Hunan 410008, PR China.
Int J Biochem Cell Biol. 2018 Sep;102:51-58. doi: 10.1016/j.biocel.2018.05.006. Epub 2018 May 16.
Lymphangiogenesis has been regarded as a physiological response to pathologic stimuli. The abnormal proliferation of lymphatic endothelial cell (LECs) and lymphangiogenesis is involved in the development of lymphatic disorders. Reportedly, VEGFC/VEGFR3 plays a key role in lymphangiogenesis; moreover, VEGFC/VEGFR3 exerts their cellular effects through activation of Ca signaling in several cell types. Herein, we demonstrated that VEGFC significantly up-regulated LEC proliferation through VEGFR3; moreover, VEGFC/VEGFR3 induced Ca signaling activation. By using online tools, miR-128 and miR-3916 were predicted as candidate upstream miRNAs which might target VEGFC/VEGFR3. As verified using Immunoblotting assays, miR-128 significantly regulated the protein levels of VEGFC/VEGFR3, whereas miR-3916 only slightly modulated VEGFC and VEGFR3 proteins. Contrary to VEGFC, miR-128 overexpression remarkably suppressed LEC proliferation, Ca release and ERK1/2-Akt signaling; moreover, the effect of VEGFC could be partially attenuated by miR-128. In summary, miR-128 interacts with the 3'-UTR of VEGFC and VEGFR3 to inhibit their expression, thus suppressing LEC proliferation through Ca and ERK1/2-Akt signaling. Taken together, we provided novel experimental basis for miRNA-regulated LEC proliferation through Ca signaling.
淋巴管生成被认为是对病理刺激的生理反应。淋巴内皮细胞（LEC）的异常增殖和淋巴管生成参与了淋巴疾病的发展。据报道，VEGFC/VEGFR3 在淋巴管生成中发挥关键作用；此外，VEGFC/VEGFR3 通过在几种细胞类型中激活 Ca 信号传导来发挥其细胞作用。在此，我们通过 VEGFR3 证明了 VEGFC 可显著上调 LEC 的增殖；此外，VEGFC/VEGFR3 诱导 Ca 信号激活。通过使用在线工具，miR-128 和 miR-3916 被预测为可能靶向 VEGFC/VEGFR3 的候选上游 miRNA。通过免疫印迹分析验证，miR-128 显著调节 VEGFC/VEGFR3 的蛋白水平，而 miR-3916 仅轻微调节 VEGFC 和 VEGFR3 蛋白。与 VEGFC 相反，miR-128 过表达可显著抑制 LEC 的增殖、Ca 释放和 ERK1/2-Akt 信号传导；此外，miR-128 可部分减弱 VEGFC 的作用。总之，miR-128 与 VEGFC 和 VEGFR3 的 3'-UTR 相互作用，抑制其表达，从而通过 Ca 和 ERK1/2-Akt 信号传导抑制 LEC 的增殖。综上所述，我们为 miRNA 通过 Ca 信号调节 LEC 增殖提供了新的实验依据。
