Zhao Yao, Cai Chenchen, Zhang Miaomiao, Shi Lubing, Wang Jiwei, Zhang Haoliang, Ma Ping, Li Shibao
Department of Clinical Laboratory, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, China.
Medical Technology School of Xuzhou Medical University, Xuzhou, 221004, China.
J Cancer Res Clin Oncol. 2021 Jul;147(7):2013-2023. doi: 10.1007/s00432-021-03618-2. Epub 2021 Mar 27.
Ephrin-A2, a member of the Eph receptor subgroup, is used in diagnosing and determining the prognosis of prostate cancer. However, the role of ephrin-A2 in prostate cancer is remains elusive.
We established stable clones overexpressing or silencing ephrin-A2 from prostate cancer cells. Then, CCK-8 was used in analyzing the proliferation ability of cells. CD31 staining was used in evaluating angiogenesis. Migration and invasion assay were conducted in vivo and in vitro. The expression of EMT-related markers was evaluated in prostate cancer cells through Western blotting.
We revealed that the ectopic expression of ephrin-A2 in prostate cancer cells facilitated cell migration and invasion in vitro and promoted tumor metastasis and angiogenesis in vivo and that the silencing of ephrin-A2 completely reversed this effect. Although ephrin-A2 did not affect tumor cell proliferation in vitro, ephrin-A2 significantly promoted primary tumor growth in vivo. Furthermore, to determine the biological function of ephrin-A2, we assayed the expression of EMT-related markers in stable-established cell lines. Results showed that the overexpression of ephrin-A2 in prostate cancer cells down-regulated the expression of epithelial markers (ZO-1, E-cadherin, and claudin-1) and up-regulated the expression of mesenchymal markers (N-cadherin, β-catenin, vimentin, Slug, and Snail), but the knocking out of ephrin-A2 opposed the effects on the expression of EMT markers.
These findings indicate that ephrin-A2 promotes prostate cancer metastasis by enhancing angiogenesis and promoting EMT and may be a potentially therapeutic target in metastatic prostate cancer.
Ephrin-A2 是 Eph 受体亚组的成员之一，可用于前列腺癌的诊断和预后判定。然而，ephrin-A2 在前列腺癌中的作用仍不清楚。
我们从前列腺癌细胞中建立了过表达或沉默 ephrin-A2 的稳定克隆。然后，使用 CCK-8 分析细胞的增殖能力。采用 CD31 染色评估血管生成。在体内和体外进行迁移和侵袭实验。通过蛋白质印迹法评估前列腺癌细胞中 EMT 相关标志物的表达。
我们发现，前列腺癌细胞中 ephrin-A2 的异位表达促进了体外细胞迁移和侵袭，促进了体内肿瘤转移和血管生成，而 ephrin-A2 的沉默完全逆转了这种作用。虽然 ephrin-A2 在体外不影响肿瘤细胞增殖，但 ephrin-A2 在体内显著促进原发性肿瘤生长。此外，为了确定 ephrin-A2 的生物学功能，我们检测了稳定建立的细胞系中 EMT 相关标志物的表达。结果显示，前列腺癌细胞中 ephrin-A2 的过表达下调了上皮标志物（ZO-1、E-钙黏蛋白和紧密连接蛋白-1）的表达，上调了间充质标志物（N-钙黏蛋白、β-连环蛋白、波形蛋白、锌指蛋白 Slug 和锌指蛋白 Snail）的表达，但敲除 ephrin-A2 则对抗了对 EMT 标志物表达的影响。
这些发现表明，ephrin-A2 通过增强血管生成和促进 EMT 促进前列腺癌转移，可能是转移性前列腺癌的潜在治疗靶点。
