环状 RNA SMARCA5 通过 miR-432/PDCD10 轴促进前列腺癌的进展。

CircSMARCA5 Facilitates the Progression of Prostate Cancer Through miR-432/PDCD10 Axis.

机构信息

Department of Urology Surgery, the Fourth Hospital of Hebei Medical University, Shijiazhuang, China.

出版信息

Cancer Biother Radiopharm. 2021 Feb;36(1):70-83. doi: 10.1089/cbr.2019.3490. Epub 2020 May 14.

DOI:10.1089/cbr.2019.3490

Abstract

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.

摘要

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。