Department of Urology, The Second Affiliated Hospital of Hainan Medical University, Haikou, P.R. China.
DNA Cell Biol. 2024 May;43(5):245-257. doi: 10.1089/dna.2023.0405. Epub 2024 Mar 15.
Clear cell renal cell carcinoma (ccRCC) is a malignant tumor of kidney epithelial cells, one of the most common tumors in the world. Transforming growth factor beta (TGFβ)1 is a crucial factor that induces epithelial-mesenchymal transition (EMT) in cancer cells. is a microRNA that is considered a tumor suppressor. However, the role and mechanism of in TGFβ1-induced ccRCC cells are not fully understood. This study investigated the roles of and its target gene in regulating EMT in ccRCC development. 786-0 and Caki-1cells were treated with TGFβ1 to induce EMT. The levels of and TGFβ2 were determined by quantitative real-time polymerase chain reaction and Western blotting. The progression of EMT was evaluated by E-cadherin detection by immunofluorescence, and E-cadherin, N-cadherin, and vimentin detection by Western blotting. Furthermore, migration and invasion capacities were assessed using a Transwell system. The direct binding of with the target gene was confirmed by dual luciferase reporter gene assay. Results indicated that TGFβ1 treatment decreased the protein abundance of E-cadherin while increasing the protein expression of N-cadherin and vimentin, indicating TGFβ1-induced EMT was constructed successfully. Moreover, TGFβ1 treatment repressed the expression of . mimics reversed the effect of TGFβ1 on the migration, invasion, and expression of E-cadherin, N-cadherin, and vimentin. The directly binds with the 3' untranslated region of mRNA and suppresses its expression. Furthermore, TGFβ2 overexpression abrogated the above changes regulated by mimics. Taken together, inhibited TGFβ1-induced EMT by suppressing the migration and invasion of ccRCC cells via directly targeting gene expression.
透明细胞肾细胞癌 (ccRCC) 是一种肾脏上皮细胞的恶性肿瘤，是世界上最常见的肿瘤之一。转化生长因子β (TGFβ)1 是诱导癌细胞上皮-间充质转化 (EMT) 的关键因素。是一种被认为是肿瘤抑制因子的 microRNA。然而，在 TGFβ1 诱导的 ccRCC 细胞中，的作用和机制尚不完全清楚。本研究探讨了及其靶基因在调节 EMT 中的作用在 ccRCC 发展中。用 TGFβ1 处理 786-0 和 Caki-1 细胞诱导 EMT。通过实时定量聚合酶链反应和 Western blot 测定 和 TGFβ2 的水平。通过免疫荧光检测 E-钙黏蛋白，Western blot 检测 E-钙黏蛋白、N-钙黏蛋白和波形蛋白来评估 EMT 的进展。此外，使用 Transwell 系统评估迁移和侵袭能力。通过双荧光素酶报告基因检测证实了与靶基因 的直接结合。结果表明，TGFβ1 处理降低了 E-钙黏蛋白的蛋白丰度，同时增加了 N-钙黏蛋白和波形蛋白的蛋白表达，表明成功构建了 TGFβ1 诱导的 EMT。此外，TGFβ1 处理抑制了的表达。模拟物逆转了 TGFβ1 对迁移、侵袭和 E-钙黏蛋白、N-钙黏蛋白和波形蛋白表达的影响。直接与 mRNA 的 3'非翻译区结合并抑制其表达。此外，TGFβ2 过表达消除了模拟物调节的上述变化。总之，通过直接靶向 基因表达，抑制 ccRCC 细胞的迁移和侵袭，抑制 TGFβ1 诱导的 EMT。
