Yang Qian, Ni Laichao, Imani Saber, Xiang Zhangqiang, Hai Rui, Ding Ruilin, Fu Shaozhi, Wu Jing Bo, Wen Qinglian
Department of Oncology, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, People's Republic of China.
Nuclear Medicine and Molecular Imaging Key Laboratory of Sichuan Province, Department of Nuclear Medicine, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, People's Republic of China.
Cancer Manag Res. 2020 Jun 24;12:4937-4948. doi: 10.2147/CMAR.S252181. eCollection 2020.
Anlotinib is a highly potent multi-target tyrosine kinase inhibitor, with very good anti-tumor activity against a variety of solid tumors. However, its effect on colorectal cancer (CRC) is not yet clearly understood. The objective of this study was to investigate the anti-tumor effect and underlying mechanism of anlotinib in the pathogenesis of CRC.
Effects of anlotinib on CT26 cells proliferation and microvessel formation in endothelial cells were determined by MTT assay and tube formation assay. Cell migration and invasion were analyzed by using the wound healing assay and transwell assay. Cell cycle and apoptosis were detected by flow cytometry. A CRC xenograft mouse model was used for conducting in-vivo studies to verify the effect of anlotinib. The expression of Ki-67 and CD31 in the tumor tissue was detected by immunohistochemistry and protein expression was measured by Western blot.
In-vitro studies revealed that anlotinib inhibited the proliferation, migration, and invasion of CT26 cells and the tube formation of HUVECs in a dose-dependent manner. Anlotinib also significantly induced cell apoptosis and G2/M arrest. It effectively inhibited tumor growth and prolonged survival time in the CRC xenograft mouse model. Immunohistochemical analysis of the tumor tissue revealed that anlotinib downregulated CD31 and Ki-67 which are the biomarkers of microvessel density and proliferation. Furthermore, anlotinib was able to inhibit the activation of VEGFR-2/AKT and FGFR, PDGFRβ and their downstream signaling ERK.
The findings of the present study suggested that anlotinib suppressed cell proliferation and angiogenesis via inhibition of AKT/ERK signaling pathway in colorectal cancer and could be a novel therapeutic strategy for treatment of CRC.
安罗替尼是一种高效的多靶点酪氨酸激酶抑制剂，对多种实体瘤具有良好的抗肿瘤活性。然而，其对结直肠癌（CRC）的作用尚未完全明确。本研究的目的是探讨安罗替尼在CRC发病机制中的抗肿瘤作用及其潜在机制。
采用MTT法和管腔形成试验测定安罗替尼对CT26细胞增殖和内皮细胞微血管形成的影响。采用伤口愈合试验和Transwell试验分析细胞迁移和侵袭能力。通过流式细胞术检测细胞周期和凋亡情况。利用CRC异种移植小鼠模型进行体内研究，以验证安罗替尼的效果。通过免疫组织化学检测肿瘤组织中Ki-67和CD31的表达，并通过蛋白质免疫印迹法测定蛋白表达水平。
体外研究表明，安罗替尼以剂量依赖性方式抑制CT26细胞的增殖、迁移和侵袭以及人脐静脉内皮细胞（HUVECs）的管腔形成。安罗替尼还显著诱导细胞凋亡和G2/M期阻滞。在CRC异种移植小鼠模型中，它有效抑制肿瘤生长并延长生存时间。对肿瘤组织的免疫组织化学分析显示，安罗替尼下调了作为微血管密度和增殖生物标志物的CD31和Ki-67。此外，安罗替尼能够抑制血管内皮生长因子受体-2（VEGFR-2）/蛋白激酶B（AKT）以及成纤维细胞生长因子受体（FGFR）、血小板衍生生长因子受体β（PDGFRβ）及其下游信号通路细胞外信号调节激酶（ERK）的激活。
本研究结果表明，安罗替尼通过抑制结直肠癌中的AKT/ERK信号通路来抑制细胞增殖和血管生成，可能是一种治疗CRC的新型治疗策略。
