Preclinical Models and New Therapeutic Agents Unit, Department of Research and Advanced Technologies, IRCCS Regina Elena National Cancer Institute, Rome, Italy.
Unit of Cell and Developmental Biology, Department of Biology, University of Pisa, Pisa, Italy.
J Exp Clin Cancer Res. 2020 Jun 5;39(1):103. doi: 10.1186/s13046-020-01604-z.
Understanding the signalling pathways involved in angiogenesis, and developing anti-angiogenic drugs are one of the major focuses on cancer research. Herein, we assessed the effect of CPTH6, a lysine acetyltransferase inhibitor and anti-tumoral compound, on angiogenesis-related properties of both endothelial and cancer cells.
The in vitro effect of CPTH6 on protein acetylation and anti-angiogenic properties on endothelial and lung cancer cells was evaluated via wound healing, trans-well invasion and migration, tube formation, immunoblotting and immunofluorescence. Matrigel plug assay, zebrafish embryo and mouse xenograft models were used to evaluate in vivo anti-angiogenic effect of CPTH6.
CPTH6 impaired in vitro endothelial angiogenesis-related functions, and decreased the in vivo vascularization both in mice xenografts and zebrafish embryos. Mechanistically, CPTH6 reduced α-tubulin acetylation and induced accumulation of acetylated microtubules in the perinuclear region of endothelial cells. Interestingly, CPTH6 also affected the angiogenesis-related properties of lung cancer cells, and conditioned media derived from CPTH6-treated lung cancer cells impaired endothelial cells morphogenesis. CPTH6 also modulated the VEGF/VEGFR2 pathway, and reshaped cytoskeletal organization of lung cancer cells. Finally, anti-migratory effect of CPTH6, dependent on α-tubulin acetylation, was also demonstrated by genetic approaches in lung cancer cells.
Overall, this study indicates that α-tubulin acetylation could play a role in the anti-angiogenic effect of CPTH6 and, more in general, it adds information to the role of histone acetyltransferases in tumor angiogenesis, and proposes the inhibition of these enzymes as an antiangiogenic therapy of cancer.
了解参与血管生成的信号通路，并开发抗血管生成药物是癌症研究的主要焦点之一。在此，我们评估了赖氨酸乙酰转移酶抑制剂和抗肿瘤化合物 CPTH6 对内皮细胞和肺癌细胞的血管生成相关特性的影响。
通过划痕实验、Transwell 侵袭和迁移实验、管形成实验、免疫印迹和免疫荧光实验评估 CPTH6 对内皮细胞和肺癌细胞蛋白乙酰化和抗血管生成特性的体外作用。利用 Matrigel plugs 实验、斑马鱼胚胎和小鼠异种移植模型评估 CPTH6 的体内抗血管生成作用。
CPTH6 损害了内皮细胞体外血管生成相关功能，并减少了小鼠异种移植和斑马鱼胚胎中的体内血管生成。机制上，CPTH6 降低了α-微管蛋白乙酰化水平，并诱导内皮细胞的核周区域中乙酰化微管的积累。有趣的是，CPTH6 还影响了肺癌细胞的血管生成相关特性，并且 CPTH6 处理的肺癌细胞条件培养基损害了内皮细胞的形态发生。CPTH6 还调节了 VEGF/VEGFR2 通路，并重塑了肺癌细胞的细胞骨架组织。最后，通过肺癌细胞的遗传方法证明了 CPTH6 的抗迁移作用依赖于α-微管蛋白乙酰化。
总体而言，这项研究表明α-微管蛋白乙酰化可能在 CPTH6 的抗血管生成作用中发挥作用，并且更广泛地说，它增加了组蛋白乙酰转移酶在肿瘤血管生成中的作用的信息，并提出抑制这些酶作为癌症的抗血管生成治疗。
