Department of Clinical Laboratory, Jiangsu Cancer Hospital and Jiangsu Institute of Cancer Research and the Affiliated Cancer Hospital of Nanjing Medical University, No. 42 Baiziting Road, Nanjing, 210009, People's Republic of China.
Department of Surgery, Jiangsu Cancer Hospital and Jiangsu Institute of Cancer Research and the Affiliated Cancer Hospital of Nanjing Medical University, Nanjing, 210009, People's Republic of China.
Breast Cancer Res Treat. 2023 Nov;202(2):397-408. doi: 10.1007/s10549-023-07090-z. Epub 2023 Aug 28.
Overactivated neddylation is considered to be a common event in cancer. Long non-coding RNAs (lncRNAs) can regulate cancer development by mediating post-translational modifications. However, the role of lncRNA in neddylation modification remains unclear.
LncRNA cytochrome P450 family 1 subfamily B member 1 antisense RNA 1 (CYP1B1-AS1) expression in breast cancer tissues was evaluated by RT-PCR and TCGA BRCA data. Gain and loss of function experiments were performed to explore the role of CYP1B1-AS1 in breast cancer cell proliferation and apoptosis in vitro and in vivo. Luciferase assay, CHIP-qPCR assay, transcriptome sequencing, RNA-pulldown assay, mass spectrometry, RIP-PCR and Western blot were used to investigate the regulatory factors of CYP1B1-AS1 expression and the molecular mechanism of CYP1B1-AS1 involved in neddylation modification.
We found that CYP1B1-AS1 was down-regulated in breast cancer tissues and correlated with prognosis. In vivo and in vitro functional experiments confirmed that CYP1B1-AS1 inhibited cell proliferation and induced apoptosis. Mechanistically, CYP1B1-AS1 was regulated by the transcription factor, forkhead box O1 (FOXO1), and could be upregulated by inhibiting the PI3K/FOXO1 pathway. Moreover, CYP1B1-AS1 bound directly to NEDD8 activating enzyme E1 subunit 1 (NAE1) to regulate protein neddylation.
This study reports for the first time that CYP1B1-AS1 inhibits protein neddylation to affect breast cancer cell proliferation, which provides a new strategy for the treatment of breast cancer by lncRNA targeting neddylation modification.
过度激活的 neddylation 被认为是癌症的一个共同事件。长链非编码 RNA（lncRNA）可以通过介导翻译后修饰来调节癌症的发展。然而，lncRNA 在 neddylation 修饰中的作用尚不清楚。
通过 RT-PCR 和 TCGA BRCA 数据评估乳腺癌组织中细胞色素 P450 家族 1 亚家族 B 成员 1 反义 RNA 1（CYP1B1-AS1）的表达。通过体外和体内的 gain 和 loss of function 实验来研究 CYP1B1-AS1 在乳腺癌细胞增殖和凋亡中的作用。利用荧光素酶报告基因检测、CHIP-qPCR 检测、转录组测序、RNA 下拉实验、质谱分析、RIP-PCR 和 Western blot 实验来研究 CYP1B1-AS1 表达的调节因子和 CYP1B1-AS1 参与 neddylation 修饰的分子机制。
我们发现 CYP1B1-AS1 在乳腺癌组织中下调，并与预后相关。体内和体外功能实验证实 CYP1B1-AS1 抑制细胞增殖并诱导细胞凋亡。机制上，CYP1B1-AS1 受转录因子叉头框 O1（FOXO1）的调控，并可通过抑制 PI3K/FOXO1 通路而上调。此外，CYP1B1-AS1 直接与 NEDD8 激活酶 E1 亚基 1（NAE1）结合，以调节蛋白质 neddylation。
本研究首次报道 CYP1B1-AS1 抑制蛋白质 neddylation 以影响乳腺癌细胞增殖，为通过 lncRNA 靶向 neddylation 修饰治疗乳腺癌提供了新策略。
