时钟基因 NR1D1 可能成为膀胱癌治疗的新靶点。
Clock gene NR1D1 might be a novel target for the treatment of bladder cancer.
机构信息
Department of Urology, Institute of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan Province, China; Department of Urology, Three Gorges Hospital, Chongqing University, Wanzhou, Chongqing, China.
Department of Urology, Institute of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan Province, China.
出版信息
Urol Oncol. 2023 Jul;41(7):327.e9-327.e18. doi: 10.1016/j.urolonc.2023.04.021. Epub 2023 May 18.
PURPOSE
To explore the role of circadian clock gene NR1D1 (REV-erbα) in bladder cancer (BC).
METHODS
Firstly, the association of NR1D1 level with clinical characteristics and prognosis was investigated among patients diagnosed with BC. Secondly, CCK-8, transwell, and colony formation experiments were performed among BC cells treated with Rev-erbα agonist (SR9009), as well as lentivirus and siRNA, for which NR1D1 were overexpressed (OE) and knocked down (KD), respectively. Thirdly, cell cycle and apoptosis were tested by flowcytometry. PI3K/AKT/mTOR pathway proteins were determined in OE-NR1D1 cells. Finally, OE-NR1D1 and OE-Control BC cells were subcutaneously implanted in BALB/c nude mice. The tumor size and protein levels were compared between groups. A P < 0.05 was considered as statistically significant.
RESULTS
Patients with NR1D1 positive status had a longer disease-free survival than those with negative expression. The cell viability, migration, and colony formation of BC cells after treated with SR9009 were significantly suppressed. OE-NR1D1 cells had obviously inhibited cell viability, migration, and colony formation, while those were found strengthened in KD-NR1D1 cells. Besides, KD-NR1D1 cells were observed with a lower proportion of dead cells and G0/G1 cells, but a higher ratio of G2/M. The changes of p-AKT, p-S6, p-4EBP1, and FASN involved in PI3K/AKT/mTOR pathway were detected in OE- and KD-NR1D1 BC cells. Finally, in vivo data demonstrated that overexpression of NR1D1 suppressed the tumorigenicity of BC cells.
CONCLUSION
NR1D1 played a role of tumor suppressor and it might become a novel target for the treatment of BC.
目的
探讨昼夜节律钟基因 NR1D1（REV-erbα）在膀胱癌（BC）中的作用。
方法
首先，在诊断为 BC 的患者中研究 NR1D1 水平与临床特征和预后的关系。其次，用 Rev-erbα 激动剂（SR9009）处理 BC 细胞，进行 CCK-8、transwell 和集落形成实验，以及用慢病毒和 siRNA 分别过表达（OE）和敲低（KD）NR1D1。然后，用流式细胞术检测细胞周期和细胞凋亡。在 OE-NR1D1 细胞中测定 PI3K/AKT/mTOR 通路蛋白。最后，将 OE-NR1D1 和 OE-Control BC 细胞皮下植入 BALB/c 裸鼠。比较各组肿瘤大小和蛋白水平。P < 0.05 为差异有统计学意义。
结果
NR1D1 阳性患者的无病生存期长于阴性表达患者。BC 细胞经 SR9009 处理后，细胞活力、迁移和集落形成明显受到抑制。OE-NR1D1 细胞的细胞活力、迁移和集落形成明显受到抑制，而 KD-NR1D1 细胞则明显增强。此外，KD-NR1D1 细胞的死亡细胞和 G0/G1 细胞比例较低，而 G2/M 细胞比例较高。OE 和 KD-NR1D1 BC 细胞中检测到 PI3K/AKT/mTOR 通路相关的 p-AKT、p-S6、p-4EBP1 和 FASN 的变化。最后，体内数据表明，NR1D1 的过表达抑制了 BC 细胞的致瘤性。
结论
NR1D1 发挥抑癌作用，可能成为治疗 BC 的新靶点。
Zotero 插件