Department of Urology, Chongqing University Three Gorges Hospital, Chongqing, China.
Department of Scientific Research and Foreign Affairs, Chongqing University Three Gorges Hospital, Chongqing, China.
Investig Clin Urol. 2022 Sep;63(5):577-588. doi: 10.4111/icu.20220153.
This research concentrated on the biological effects and special mechanism of circ_0003221 in bladder cancer (BLCA).
The level quantification by reverse transcription-quantitative polymerase chain reaction was administrated for circ_0003221, microRNA-892b (miR-892b) and 24-dehydrocholesterol reductase (DHCR24). The biological behaviors were assessed by EDU assay and colony formation assay for proliferation, and transwell assay for cell motility. Glycolytic metabolism was tested using the commercial kits. DHCR24 protein level and cell markers were measured through western blot. The analysis of interaction potential was conducted via dual-luciferase reporter assay and pull-down assay. Circ_0003221 was implemented via tumor xenograft assay .
Abnormal circ_0003221 upregulation was affirmed in BLCA. BLCA cell proliferation, motility and glycolysis were impeded after circ_0003221 level was knocked down. MiR-892b was identified as a target for circ_0003221. Reduction of miR-892b relieved si-circ_0003221-induced anti-tumor response in BLCA cells. In addition, miR-892b targeted DHCR24 and circ_0003221/miR-892b could regulate the level of DHCR24. The effects of si-circ_0003221 were also counteracted by DHCR24 overexpression.
The current evidence elucidated circ_0003221 targeted miR-892b to elevate the DHCR24 level, thus accelerating cell development and glycolytic metabolism of BLCA cells.
本研究集中探讨环状 RNA （circRNA）_0003221 在膀胱癌（BLCA）中的生物学效应及特殊作用机制。
采用逆转录定量聚合酶链反应（RT-qPCR）对 circ_0003221、微小 RNA-892b（miR-892b）和 24-脱氢胆固醇还原酶（DHCR24）进行水平定量。通过 EDU 检测和集落形成实验评估增殖能力，Transwell 检测评估细胞迁移能力。使用商业试剂盒检测糖酵解代谢。通过 Western blot 检测 DHCR24 蛋白水平和细胞标志物。通过双荧光素酶报告基因检测和下拉实验分析相互作用潜力。通过肿瘤异种移植实验研究 circ_0003221。
BLCA 中存在异常的 circ_0003221 上调。circ_0003221 水平降低后，BLCA 细胞增殖、迁移和糖酵解受到抑制。miR-892b 被鉴定为 circ_0003221 的靶基因。降低 miR-892b 可缓解 BLCA 细胞中 si-circ_0003221 诱导的抗肿瘤反应。此外，miR-892b 靶向 DHCR24，circ_0003221/miR-892b 可调节 DHCR24 水平。DHCR24 过表达也可逆转 si-circ_0003221 的作用。
目前的证据表明，circ_0003221 通过靶向 miR-892b 来提高 DHCR24 水平，从而加速 BLCA 细胞的发育和糖酵解代谢。
