Department of Urology Surgery, the Fourth Hospital of Hebei Medical University, Shijiazhuang, China.
Cancer Biother Radiopharm. 2021 Feb;36(1):70-83. doi: 10.1089/cbr.2019.3490. Epub 2020 May 14.
Circular RNAs (circRNAs) have been reported to be implicated in the pathogenesis of prostate cancer (PCa). Herein, the authors explore the role and molecular mechanism of circRNA SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a, member 5 (circSMARCA5) in PCa. The levels of circSMARCA5, SMARCA5, miR-432, and programmed cell death 10 (PDCD10) were determined by quantitative real-time polymerase chain reaction (qRT-PCR). The circular structure and stability of circSMARCA5 were validated by qRT-PCR using Oligo dT primer, transcriptional inhibitor actinomycin D, or RNase R treatment, respectively. Cell proliferation, migration, invasion, epithelial/mesenchymal transition (EMT), and glycolysis were detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), transwell migration and invasion assays, Western blot assay, and Glucose or Lactate Detection Kit, respectively. The target relationship between miR-432 and circSMARCA5 or PDCD10 was validated by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Western blot was performed to detect the protein expression of PDCD10 in PCa cells. CircSMARCA5 was aberrantly upregulated, and was a circular and stable RNA in PCa cells. CircSMARCA5 accelerated the proliferation, metastasis, and glycolysis of PCa cells. MiR-432 was a direct target of circSMARCA5, and circSMARCA5 accelerated the development of PCa through miR-432 in PCa cells. PDCD10 was a direct target of miR-432, and PDCD10 addition reversed the inhibitory effects of miR-432 accumulation on the proliferation, metastasis, and glycolysis of PCa cells. CircSMARCA5 upregulated the expression of PDCD10 through sponging miR-432 in PCa cells. CircSMARCA5 deteriorated PCa through the miR-432/PDCD10 axis. CircSMARCA5/miR-432/PDCD10 axis might be an underlying therapeutic target for PCa treatment.
环状 RNA（circRNA）已被报道参与前列腺癌（PCa）的发病机制。在此，作者研究了环状 RNA SWI/SNF 相关、基质相关、肌动蛋白依赖性染色质调节剂亚家族 A、成员 5（circSMARCA5）在 PCa 中的作用和分子机制。通过实时定量聚合酶链反应（qRT-PCR）测定 circSMARCA5、SMARCA5、miR-432 和程序性细胞死亡 10（PDCD10）的水平。通过使用 Oligo dT 引物、转录抑制剂放线菌素 D 或核糖核酸酶 R 处理，分别通过 qRT-PCR 验证 circSMARCA5 的环状结构和稳定性。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐（MTT）、Transwell 迁移和侵袭试验、Western blot 试验和葡萄糖或乳酸检测试剂盒分别检测细胞增殖、迁移、侵袭、上皮/间充质转化（EMT）和糖酵解。通过双荧光素酶报告基因检测和 RNA 免疫沉淀（RIP）试验验证 miR-432 与 circSMARCA5 或 PDCD10 的靶关系。通过 Western blot 检测 PCa 细胞中 PDCD10 的蛋白表达。circSMARCA5 在 PCa 细胞中异常上调，是一种环状和稳定的 RNA。circSMARCA5 加速了 PCa 细胞的增殖、转移和糖酵解。miR-432 是 circSMARCA5 的直接靶标，circSMARCA5 通过 miR-432 在 PCa 细胞中加速了 PCa 的发展。PDCD10 是 miR-432 的直接靶标，PDCD10 的添加逆转了 miR-432 积累对 PCa 细胞增殖、转移和糖酵解的抑制作用。circSMARCA5 通过海绵吸附 miR-432 在 PCa 细胞中上调 PDCD10 的表达。circSMARCA5 通过 miR-432/PDCD10 轴恶化 PCa。circSMARCA5/miR-432/PDCD10 轴可能是 PCa 治疗的潜在治疗靶点。
