Gao Pan, Liu Hongtao, Yang Zhenzhen, Hui Yiran, Shi Zhuangzhuang, Yang Zhen, Song Min, Yao Menghui, Fan Wenfei, Yang Jinhua, Hao Yibin, Fan Tianli
People's Hospital of Zhengzhou, Zhengzhou, Henan, 450001, People's Republic of China.
Department of Pharmacology, School of Basic Medicine, Zhengzhou University, Zhengzhou, Henan, 450001, People's Republic of China.
Onco Targets Ther. 2021 Feb 4;14:785-793. doi: 10.2147/OTT.S290564. eCollection 2021.
Increasing evidence has demonstrated that animal models are imperative to investigate the potential molecular mechanism of metastasis and discover anti-metastasis drugs; however, efficient animal models to unveil the underlying mechanisms of metastasis in esophageal squamous cell carcinoma (ESCC) are limited.
ESCC cell EC9706 with high invasiveness was screened by repeated Transwell assays. Its biological characteristics were identified by flow cytometry as well as by the wound healing and CCK-8 assays. Besides, the levels of epithelial-mesenchymal transition-related markers were examined using Western blotting. Parental (EC9706-I) and subpopulation (EC9706-I) cells were employed to establish the renal capsule model. Next, the tumor growth was detected by a live animal imaging system, and hematoxylin and eosin staining was applied to evaluate the metastatic status in ESCC.
EC9706-I cells showed rapid proliferation ability, S phase abundance, and high invasive ability; obvious upregulation in N-cadherin, Snail, Vimentin, and Bit1; and downregulation in E-cadherin. EC9706-I cells were less sensitive to the chemotherapy drug 5-fluorouracil than EC9706-I cells; however, both cell lines reached a tumorigenesis rate of 100% in the renal capsule model. The live animal imaging system revealed that the tumors derived from EC9706-I cells grew more slowly than those from EC9706-I cells at weeks 3-14. The EC9706-I xenograft model displayed a spontaneous metastatic site, including kidney, heart, liver, lung, pancreas, and spleen, with a distant metastatic rate of 80%.
Our data suggested that the metastatic model was successfully established, providing a novel platform for further exploring the molecular mechanisms of metastasis in ESCC patients.
越来越多的证据表明，动物模型对于研究转移的潜在分子机制和发现抗转移药物至关重要；然而，用于揭示食管鳞状细胞癌（ESCC）转移潜在机制的有效动物模型有限。
通过反复的Transwell实验筛选出具有高侵袭性的ESCC细胞EC9706。通过流式细胞术、伤口愈合实验和CCK-8实验鉴定其生物学特性。此外，使用蛋白质免疫印迹法检测上皮-间质转化相关标志物的水平。采用亲代（EC9706-I）和亚群（EC9706-I）细胞建立肾包膜模型。接下来，通过活体动物成像系统检测肿瘤生长情况，并应用苏木精-伊红染色评估ESCC的转移状态。
EC9706-I细胞表现出快速增殖能力、S期丰度和高侵袭能力；N-钙黏蛋白、Snail、波形蛋白和Bit1明显上调；E-钙黏蛋白下调。EC9706-I细胞对化疗药物5-氟尿嘧啶的敏感性低于EC9706-I细胞；然而，在肾包膜模型中，两种细胞系的成瘤率均达到100%。活体动物成像系统显示，在第3-14周，源自EC9706-I细胞的肿瘤生长比源自EC9706-I细胞的肿瘤更缓慢。EC9706-I异种移植模型显示出一个自发转移部位，包括肾脏、心脏、肝脏、肺、胰腺和脾脏，远处转移率为80%。
我们的数据表明成功建立了转移模型，为进一步探索ESCC患者转移的分子机制提供了一个新平台。
