Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.
National Key Laboratory of Advanced Drug Delivery and Release Systems, Zhejiang University, Hangzhou, China.
Int J Immunopathol Pharmacol. 2023 Jan-Dec;37:3946320231184997. doi: 10.1177/03946320231184997.
Ten-eleven translocation 1 (TET1) is a member of the DNA demethylase family that regulates the methylation level of the genome. Dysregulation of TET1 in renal cell carcinoma (RCC) may be associated with RCC progression, but the mechanism of TET1 down-regulation in RCC is not yet known. MiR-183-5p is up-regulated in various tumor tissues and acts as an oncogene. We used Transwell and wound healing assays to test cell invasion and migration. To investigate DNA methylation, we used dot blot, which indicates TET1 enzyme activity. We verified the binding of miR-183-5p and TET1 3'-UTR (untranslated region) using dual-luciferase reporter assay. Our study demonstrated, for the first time, that miR-183-5p can directly repress TET1 expression in RCC. We observed a significant decrease in TET1 expression in RCC specimens, as reported in the literature, and a significant decrease in the concentration of 5hmC in RCC. By aligning the microRNA with a database and using the luciferase reporter gene method, we found that miR-183-5p can inhibit luciferase activity by binding to 453-459 bp of TET1 3'-UTR, leading to inhibition of TET1 expression. Furthermore, down-regulation of TET1 inhibited miR-200c expression and promoted RCC cell invasion and migration. Our findings suggest that in RCC, increased expression of miR-183-5p inhibits the expression of TET1, which in turn inhibits the expression of miR-200c and E-cadherin, both of which are associated with cell adhesion. This leads to the promotion of cell invasion and migration.
十号十一号转位 1（TET1）是 DNA 去甲基化酶家族的一员，调节基因组的甲基化水平。TET1 在肾细胞癌（RCC）中的失调可能与 RCC 的进展有关，但 RCC 中 TET1 下调的机制尚不清楚。miR-183-5p 在各种肿瘤组织中上调，作为一种癌基因发挥作用。我们使用 Transwell 和划痕愈合实验来测试细胞侵袭和迁移。为了研究 DNA 甲基化，我们使用点印迹法，这表明 TET1 酶活性。我们使用双荧光素酶报告基因检测验证了 miR-183-5p 和 TET1 3'-UTR（非翻译区）的结合。我们的研究首次表明，miR-183-5p 可以直接抑制 RCC 中的 TET1 表达。我们观察到 RCC 标本中 TET1 表达明显下降，正如文献报道的那样，RCC 中 5hmC 的浓度也明显下降。通过将 microRNA 与数据库对齐并使用荧光素酶报告基因方法，我们发现 miR-183-5p 可以通过结合 TET1 3'-UTR 的 453-459 bp 来抑制荧光素酶活性，从而抑制 TET1 的表达。此外，下调 TET1 抑制 miR-200c 的表达并促进 RCC 细胞侵袭和迁移。我们的研究结果表明，在 RCC 中，miR-183-5p 的表达增加抑制了 TET1 的表达，进而抑制了 miR-200c 和 E-cadherin 的表达，这两者都与细胞黏附有关。这导致了细胞侵袭和迁移的促进。
