Department of Urology, Zhongshan Hospital Xiamen University, Xiamen, Fujian, China.
The Third Clinical Medical College, Fujian Medical University, Xiamen, Fujian, China.
Bioengineered. 2021 Dec;12(2):11089-11107. doi: 10.1080/21655979.2021.2008217.
Circular RNAs (CircRNAs) gain importance as regulatory molecules in prostate cancer (PCa), but molecular mechanism of most circRNAs in pathogenesis of PCa remains to be studied. This study aimed to explore the role of hsa_circ_0030586 in PCa. Gene Expression Omnibus database (GSE77661) was used to screen out candidate circRNAs. Quantitative real-time PCR was used to verify the relative expressions of circRNAs, miRNAs, and genes in PCa cells. A CCK-8 assay was used to evaluate the cells' proliferation. Transwell and wound healing assay were used to determine the cells' migration and invasion. Western blotting and immunohistochemistry were used to detect the protein expression of PI3K/AKT signaling proteins and epithelial-mesenchymal transition (EMT) markers. Furthermore, a nude mice tumorigenesis experiment in vivo was conducted to determine the function of hsa_circ_0030586 on PCa. Our results showed that hsa_circ_0030586 is significantly upregulated in PCa cells (p < 0.05). Its circular structure was confirmed via agarose gel electrophoresis and Sanger sequencing. Interfering with hsa_circ_0030586 in PC3 cells inhibited cell proliferation, migration, and invasion and led to the significant upregulation of E-cadherin and the significant downregulation of p-AKT/AKT, IKKα, PIK3CB, and Twist (all p < 0.05). Conversely, the hsa_circ_003058 interference fragment combined with the transfection of a miR-145-3p inhibitor could reverse the above effects. In vivo tumorigenesis of the xenograft model confirmed that interfering with hsa_circ_0030586 suppressed tumor cell proliferation and inhibited PI3K-AKT signaling and EMT in PC3 cells. Hsa_circ_0030586 is significantly upregulated in PCa cells and may promote EMT via PI3K-AKT signaling.
环状 RNA（circRNAs）作为前列腺癌（PCa）的调控分子的重要性日益增加，但大多数 circRNAs 在 PCa 发病机制中的分子机制仍有待研究。本研究旨在探讨 hsa_circ_0030586 在 PCa 中的作用。使用基因表达综合数据库（GSE77661）筛选候选 circRNAs。使用定量实时 PCR 验证 PCa 细胞中 circRNAs、miRNAs 和基因的相对表达水平。CCK-8 测定法用于评估细胞增殖。Transwell 和划痕愈合实验用于测定细胞迁移和侵袭。Western blot 和免疫组化用于检测 PI3K/AKT 信号蛋白和上皮间质转化（EMT）标志物的蛋白表达。此外，进行体内裸鼠肿瘤发生实验以确定 hsa_circ_0030586 对 PCa 的功能。我们的结果表明，hsa_circ_0030586 在 PCa 细胞中显著上调（p<0.05）。通过琼脂糖凝胶电泳和 Sanger 测序确认其环状结构。在 PC3 细胞中干扰 hsa_circ_0030586 抑制细胞增殖、迁移和侵袭，并导致 E-钙黏蛋白的显著上调和 p-AKT/AKT、IKKα、PIK3CB 和 Twist 的显著下调（均 p<0.05）。相反，hsa_circ_003058 干扰片段与 miR-145-3p 抑制剂的转染结合可以逆转上述作用。异种移植模型的体内肿瘤发生证实，干扰 hsa_circ_0030586 抑制 PC3 细胞的肿瘤细胞增殖并抑制 PI3K-AKT 信号和 EMT。hsa_circ_0030586 在 PCa 细胞中显著上调，可能通过 PI3K-AKT 信号促进 EMT。
