1 Department of Urology, Qilu Hospital, Shandong University, Jinan, China.
2 Department of Urology, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, China.
Hum Exp Toxicol. 2019 Aug;38(8):927-937. doi: 10.1177/0960327119842241. Epub 2019 Apr 17.
To investigate the effects of EH domain containing protein 2 (EHD2) on clear cell renal cell carcinoma (ccRCC) and provide new insights for the clinical treatment of rental cancer. Forty patients (26 males and 14 females, 62.4 ± 5.7 years old) with ccRCC were selected from January 2015 to December 2016 to serve as research subjects in this study. The EHD2 protein expression in the tumor tissues and adjacent healthy tissues of ccRCC patients were detected by Western Blot assay. The cells of ccRCC cell lines RLC-310 and 786-O were divided into normal control group (control), no-load control group (pLV), EHD2 overexpression group (pLV-EHD2), and EHD2 interference group (pLV-siEHD2). The expression levels of EHD2 protein in each group of cells were detected by western blot. The cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. Wound healing assay was performed to check the cell migration ability. Transwell invasion assay was used to detect the cell invasion ability. Cell apoptosis was detected by flow cytometry. The expression level of EHD2 was significantly increased in pLV-EHD2 group and decreased in pLV-siEHD2 group compared with control group and pLV-siEHD2 group, indicating the successfully established EHD2 overexpression cell line and EHD2 RNA interference cell line. EHD2 overexpression enhanced the proliferation, invasion, and migration but inhibited the apoptosis of ccRCC cells, while EHD2 interference showed opposite functions. EHD2 interference can inhibit the development of ccRCC by inhibiting the proliferation, invasion, and migration, and EHD2 can potentially serve as a molecular target for the clinical treatment of ccRCC.
为了研究 EH 结构域包含蛋白 2（EHD2）对肾透明细胞癌（ccRCC）的影响，为肾癌的临床治疗提供新的思路。本研究选取 2015 年 1 月至 2016 年 12 月期间在我院接受治疗的 40 例 ccRCC 患者（男 26 例，女 14 例，年龄 62.4±5.7 岁）作为研究对象。采用 Western blot 法检测 ccRCC 患者肿瘤组织和癌旁正常组织中 EHD2 蛋白的表达。将 ccRCC 细胞系 RLC-310 和 786-O 的细胞分为正常对照组（对照组）、空载对照组（pLV）、EHD2 过表达组（pLV-EHD2）和 EHD2 干扰组（pLV-siEHD2）。采用 Western blot 法检测各组细胞中 EHD2 蛋白的表达水平，采用细胞计数试剂盒-8（CCK-8）法检测细胞增殖能力，采用划痕愈合实验检测细胞迁移能力，采用 Transwell 侵袭实验检测细胞侵袭能力，采用流式细胞术检测细胞凋亡。与对照组和 pLV-siEHD2 组相比，pLV-EHD2 组 EHD2 蛋白表达水平明显升高，pLV-siEHD2 组 EHD2 蛋白表达水平明显降低，表明成功建立了 EHD2 过表达细胞系和 EHD2 RNA 干扰细胞系。EHD2 过表达增强了 ccRCC 细胞的增殖、侵袭和迁移能力，但抑制了细胞凋亡，而 EHD2 干扰则表现出相反的作用。EHD2 干扰通过抑制 ccRCC 的增殖、侵袭和迁移，抑制 ccRCC 的发生发展，EHD2 可能成为 ccRCC 临床治疗的分子靶点。
