Department of Urology, Renmin Hospital of Wuhan University, Wuhan, China.
J Clin Lab Anal. 2022 Apr;36(4):e24360. doi: 10.1002/jcla.24360. Epub 2022 Mar 18.
Prostate cancer (PCa) is a frequent malignant tumor worldwide with high morbidity along with mortality. MicroRNAs (miRNAs) have been identified as key posttranscriptional modulators in diverse cancers. Herein, we purposed to explore the impacts of miR-363-3p on PCa growth, migration, infiltration along with apoptosis.
The expressions of miR-363-3p along with Dickkopf 3 (DKK3) were assessed in clinical PCa specimens. We adopted the PCa cell line PC3 and transfected it using miR-363-3p repressors or mimic. The relationship of miR-363-3p with DKK3 was verified by a luciferase enzyme reporter assay. Cell viability along with apoptosis were determined by MTT assay coupled with flow cytometry analysis. Cell migration along infiltration were detected via wound healing, as well as Transwell assays. The contents of DKK3, E-cadherin, vimentin along with N-cadherin were analyzed via Western blotting accompanied with qRT-PCR.
MiR-363-3p was found to be inversely associated with the content of DKK3 in clinical PCa specimens. Further investigations revealed that DKK3 was miR-363-3p's direct target. Besides, overexpression of miR-363-3p decreased the contents of DKK3, promoted cell viability, migration coupled with infiltration, and reduced cell apoptosis, while silencing of miR-363-3p resulted in opposite influence. Upregulation of miR-363-3p diminished E-cadherin contents but increased vimentin along with N-cadherin protein contents in PC3 cells; in contrast, miR-363-3p downregulation produced the opposite result.
Our study indicates that miR-363-3p promotes PCa growth, migration coupled with invasion while dampening apoptosis by targeting DKK3.
前列腺癌（PCa）是一种常见的恶性肿瘤，全球发病率和死亡率都很高。微小 RNA（miRNA）已被确定为多种癌症中关键的转录后调节因子。在此，我们旨在探讨 miR-363-3p 对 PCa 生长、迁移、浸润和凋亡的影响。
评估临床 PCa 标本中 miR-363-3p 和 Dickkopf 3（DKK3）的表达。我们采用 PCa 细胞系 PC3，并使用 miR-363-3p 抑制剂或模拟物进行转染。通过荧光素酶酶报告基因测定验证 miR-363-3p 与 DKK3 的关系。通过 MTT 测定结合流式细胞术分析测定细胞活力和凋亡。通过划痕愈合和 Transwell 测定检测细胞迁移和浸润。通过 Western blot 分析和 qRT-PCR 分析检测 DKK3、E-钙黏蛋白、波形蛋白和 N-钙黏蛋白的含量。
miR-363-3p 与临床 PCa 标本中 DKK3 的含量呈负相关。进一步研究表明，DKK3 是 miR-363-3p 的直接靶标。此外，过表达 miR-363-3p 降低了 DKK3 的含量，促进了细胞活力、迁移和浸润，并减少了细胞凋亡，而沉默 miR-363-3p 则产生了相反的影响。上调 miR-363-3p 降低了 PC3 细胞中 E-钙黏蛋白的含量，但增加了波形蛋白和 N-钙黏蛋白的蛋白含量；相反，下调 miR-363-3p 则产生了相反的结果。
我们的研究表明，miR-363-3p 通过靶向 DKK3 促进 PCa 的生长、迁移和浸润，同时抑制凋亡。
