外泌体 Circ-XIAP 通过调控 miR-1182/TPD52 轴促进前列腺癌多西他赛耐药。
Exosomal Circ-XIAP Promotes Docetaxel Resistance in Prostate Cancer by Regulating miR-1182/TPD52 Axis.
机构信息
College of Medical, Huanghuai University, Zhumadian, Henan, People's Republic of China.
Department of Urology, The Central Hospital of Zhumadian, Zhumadian, Henan, People's Republic of China.
出版信息
Drug Des Devel Ther. 2021 May 3;15:1835-1849. doi: 10.2147/DDDT.S300376. eCollection 2021.
BACKGROUND
Exosomal circular RNAs (circRNAs) are involved in the pathogenesis of prostate cancer (PCa) and chemotherapy resistance. This research aimed to explore the function and molecular mechanism of circRNA X-linked inhibitor of apoptosis (circ-XIAP) in docetaxel (DTX) resistance of PCa.
METHODS
The expression of circ-XIAP, microRNA-1182 (miR-1182), tumor protein D52 (TPD52) was measured by quantitative real-time polymerase chain reaction (qRT-PCR). Exosomes were detected with transmission electron microscopy (TEM). Cluster of differentiation 63 (CD63), cluster of differentiation 9 (CD9) and TPD52 protein levels were detected by Western blot (WB). FIfty percent inhibitory concentration (IC50) of DTX and cell viability were determined using Cell Counting Kit-8 (CCK-8) assay. Colony formation assay was applied to assess colony-forming ability. Cell cycle distribution and apoptosis were analyzed by flow cytometry. Transwell assay was used for measuring cell migration and invasion. Dual-reporter luciferase assay was performed to confirm the interaction between miR-1182 and circ-XIAP or TPD52. The role of circ-XIAP in vivo was confirmed via the mice xenograft model.
RESULTS
Circ-XIAP and TPD52 were upregulated and miR-1182 was downregulated in DTX-resistant PCa tissue specimens and cell lines. Circ-XIAP was also overexpressed in exosomes from DTX-resistant cells and could be transmitted via exosomes. Circ-XIAP knockdown enhanced DTX sensitivity by suppressing DTX-resistant cell proliferation, migration and invasion and inducing cell cycle arrest and apoptosis. Circ-XIAP directly targeted miR-1182, and the effects of circ-XIAP knockdown were reversed by downregulating miR-1182 in DTX-resistant cells. TPD52 was the target of miR-1182, and its upregulation weakened the promotive effect of miR-1182 on DTX sensitivity. Importantly, circ-XIAP depletion inhibited tumor growth and increased DTX sensitivity in vivo.
CONCLUSION
Exosomal circ-XIAP promoted DTX resistance of PCa by regulating miR-1182/TPD52 axis, providing a promising therapeutic target for PCa chemotherapy.
背景
外泌体环状 RNA（circRNA）参与前列腺癌（PCa）的发病机制和化疗耐药性。本研究旨在探讨 X 连锁凋亡抑制因子（circ-XIAP）在 PCa 多西他赛（DTX）耐药中的作用及其分子机制。
方法
采用实时定量聚合酶链反应（qRT-PCR）检测 circ-XIAP、微小 RNA-1182（miR-1182）和肿瘤蛋白 D52（TPD52）的表达。采用透射电子显微镜（TEM）检测外泌体。采用 Western blot（WB）检测 CD63、CD9 和 TPD52 蛋白水平。采用细胞计数试剂盒-8（CCK-8）检测 DTX 的 50%抑制浓度（IC50）和细胞活力。采用集落形成实验评估集落形成能力。采用流式细胞术分析细胞周期分布和凋亡。Transwell 实验用于检测细胞迁移和侵袭。双荧光素酶报告基因实验验证 miR-1182 与 circ-XIAP 或 TPD52 的相互作用。通过小鼠异种移植模型证实 circ-XIAP 在体内的作用。
结果
在 DTX 耐药的 PCa 组织标本和细胞系中，circ-XIAP 和 TPD52 上调，miR-1182 下调。在 DTX 耐药细胞来源的外泌体中也过表达 circ-XIAP，并且可以通过外泌体传递。circ-XIAP 敲低通过抑制 DTX 耐药细胞增殖、迁移和侵袭，诱导细胞周期阻滞和凋亡，增强 DTX 敏感性。circ-XIAP 直接靶向 miR-1182，在 DTX 耐药细胞中下调 miR-1182 可逆转 circ-XIAP 敲低的作用。TPD52 是 miR-1182 的靶基因，上调 TPD52 可减弱 miR-1182 对 DTX 敏感性的促进作用。重要的是，circ-XIAP 耗竭抑制体内肿瘤生长并增加 DTX 敏感性。
结论
外泌体 circ-XIAP 通过调节 miR-1182/TPD52 轴促进 PCa 的 DTX 耐药性，为 PCa 化疗提供了有前途的治疗靶点。
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