Chu Zewen, Shi Xin, Chen Gaoyang, He Xuejun, Qian Yayun, Wang Haibo, Tao Li, Liu Yanqing, Jiang Wei, Chen Jue
Department of Oncology, The Second People's Hospital of Taizhou Affiliated to Medical College of Yangzhou University, Yangzhou, China.
The Key of Cancer Prevention and Treatment of Yangzhou University, Yangzhou, China.
Front Pharmacol. 2021 Mar 31;12:619732. doi: 10.3389/fphar.2021.619732. eCollection 2021.
New strategies and drugs are urgently needed to improve the treatment of hepatocellular carcinoma (HCC). Vasculogenic mimicry (VM) has been elucidated being associated with the progression of HCC and anti-VM could be a promising strategy. extract (COE), a mixture of 26 compounds isolated from the Chinese Herb Vine, has been elucidated to be able to disrupt VM formation in HCC. This study aims to dissect and identify the potential targets of COE on anti-VM formation both and that are distinct from our previous study. Proteomics analysis was used to identify differential proteins in HCC cells treated with or without COE (Data are available ProteomeXchange with identifier PXD022203). Cells invasion was examined using Transwell. Matrigel was used to establish a 3-D culture condition for VM formation . RT-PCR and Western Blot were used to examine changes of mRNA and protein respectively. Clinical resected samples were applied to confirm association between VM formation and identified targets. Subcutaneous xenograft tumor model was established to observe tumor growth and VM formation . PAS-CD34 dual staining was used to detect VM . A total of 194 proteins were identified to be differentially expressed in HCC cells treated with or without COE. In the 93 down-regulated proteins EphA2 stood out to be regulated on both RNA and protein level. Disruption EphA2 using COE or NVP inhibited VM formation and decreased VM associated biomarkers. In xenograft mouse model, COE inhibited tumor growth and VM formation via down-regulating EphA2. Taken together, our results indicate that COE could be used in HCC treatment because of its promising anti-VM effect.
迫切需要新的策略和药物来改善肝细胞癌（HCC）的治疗。血管生成拟态（VM）已被阐明与HCC的进展相关，抗VM可能是一种有前景的策略。中药藤黄提取物（COE）是从藤黄中分离出的26种化合物的混合物，已被阐明能够破坏HCC中的VM形成。本研究旨在剖析并确定COE在抗VM形成方面的潜在靶点，这些靶点与我们之前的研究不同。蛋白质组学分析用于鉴定用或不用COE处理的HCC细胞中的差异蛋白（数据可在ProteomeXchange上获取，标识符为PXD022203）。使用Transwell检测细胞侵袭。使用基质胶建立用于VM形成的三维培养条件。RT-PCR和蛋白质印迹分别用于检测mRNA和蛋白质的变化。应用临床切除样本确认VM形成与鉴定出的靶点之间的关联。建立皮下异种移植肿瘤模型以观察肿瘤生长和VM形成。PAS-CD34双重染色用于检测VM。共鉴定出194种蛋白质在用或不用COE处理的HCC细胞中差异表达。在93种下调的蛋白质中，EphA2在RNA和蛋白质水平上均受到调控。使用COE或NVP破坏EphA2可抑制VM形成并降低VM相关生物标志物。在异种移植小鼠模型中，COE通过下调EphA2抑制肿瘤生长和VM形成。综上所述，我们的结果表明，由于其有前景的抗VM作用，COE可用于HCC治疗。
