Ma Jing, Chen Xu, Chen Yumo, Tao Ning, Qin Zhihai
Key Laboratory of Protein and Peptide Pharmaceuticals, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
The First Affiliated Hospital of Zhengzhou University, Zhengzhou University, Zhengzhou 450052, China.
Cancers (Basel). 2022 May 13;14(10):2406. doi: 10.3390/cancers14102406.
CAFs secrete VEGFA in the tumor microenvironment to induce angiogenesis and promote tumor growth. The downregulation of VEGFA secretion from CAFs helps block angiogenesis and exerts an anti-tumor effect. In vivo experiments showed that the angiogenesis of the tumor-bearing mice in the ligustilide group was significantly reduced. The results of MTT, tube formation, Transwell and scratch experiments showed that ligustilide did not affect the proliferation of HUVECs in a certain concentration range (<60 μM), but it inhibited the proliferation, tube formation and migration of HUVECs induced by CAFs. At this concentration, ligustilide did not inhibit CAF proliferation. The qPCR and WB results revealed that ligustilide downregulated the level of VEGFA in CAFs via the TLR4-ERK/JNK/p38 signaling pathway, and the effect was attenuated by blockers of the above molecules. Ligustilide also downregulated the autocrine VEGFA of HUVECs induced by CAFs, which inhibited angiogenesis more effectively. In addition, ligustilide inhibited glycolysis and HIF-1 expression in CAFs. Overall, ligustilide downregulated the VEGFA level in CAFs via the TLR4-ERK/JNK/p38 signaling pathway and inhibited the promotion of angiogenesis. This study provides a new strategy for the anti-tumor effect of natural active molecules, namely, blockade of angiogenesis, and provides a new candidate molecule for blocking angiogenesis in the tumor microenvironment.
癌相关成纤维细胞（CAFs）在肿瘤微环境中分泌血管内皮生长因子A（VEGFA）以诱导血管生成并促进肿瘤生长。CAFs分泌VEGFA的下调有助于阻断血管生成并发挥抗肿瘤作用。体内实验表明，藁本内酯组荷瘤小鼠的血管生成显著减少。MTT、管腔形成、Transwell和划痕实验结果表明，在一定浓度范围（<60μM）内藁本内酯不影响人脐静脉内皮细胞（HUVECs）的增殖，但它抑制了CAFs诱导的HUVECs的增殖、管腔形成和迁移。在此浓度下，藁本内酯不抑制CAF的增殖。定量聚合酶链反应（qPCR）和蛋白质免疫印迹（WB）结果显示，藁本内酯通过Toll样受体4（TLR4）-细胞外信号调节激酶（ERK）/应激活化蛋白激酶（JNK）/p38信号通路下调CAFs中VEGFA的水平，并且上述分子的阻断剂减弱了该作用。藁本内酯还下调了CAFs诱导的HUVECs的自分泌VEGFA，从而更有效地抑制血管生成。此外，藁本内酯抑制了CAFs中的糖酵解和缺氧诱导因子-1（HIF-1）表达。总体而言，藁本内酯通过TLR4-ERK/JNK/p38信号通路下调CAFs中VEGFA的水平并抑制血管生成的促进作用。本研究为天然活性分子的抗肿瘤作用提供了一种新策略，即阻断血管生成，并为阻断肿瘤微环境中的血管生成提供了一种新的候选分子。
