Li Ying, Wang Yu, Gao Jun, Xu Weiran, Wang Yingkai, Zhang Fan
Department of Gastroenterology, The First Hospital of Jilin University, Changchun, China.
Endoscopy Center, The First Hospital of Jilin University, Changchun, China.
Iran J Biotechnol. 2025 Apr 1;23(2):e4071. doi: 10.30498/ijb.2025.504573.4071. eCollection 2025 Apr.
This study aimed to investigate the role of lncRNA ZFAS1 in gastric cancer progression, focusing on its regulation by SP1 and its impact on the AKT/mTOR signaling pathway. By exploring ZFAS1's effects on cell proliferation, migration, and apoptosis, we sought to uncover its molecular mechanisms and potential as a therapeutic target.
We evaluated ZFAS1 expression in gastric cancer cells (SGC7901) by RT-qPCR and compared it with GES-1 cells. The LnCAR database provided insight into ZFAS1 levels in STAD compared to normal tissue. To knockdown ZFAS1 in SGC7901 cells, we transfected the cells with si-ZFAS1 #1-3 (small interfering RNAs targeting ZFAS1), and si-ZFAS1-2 was found to have the highest knockdown efficiency. Then, the effect of ZFAS1 knockdown on cell invasion, migration and proliferation was evaluated using transwell invasion, wound healing assays, CCK8 and flow cytometry. In addition, ZFAS1 promoter regions were examined using the JASPAR database and subsequent ChIP assays to understand SP1 transcription factor binding. The effect of ZFAS1 on the AKT/mTOR pathway was clarified using Western blotting.
SGC7901 cells were shown to have increased ZFAS1 expression, which was linked to a poor prognosis for gastric cancer. Knockdown of ZFAS1 in SGC7901 cells inhibited cell invasion, migration and proliferation and induced apoptosis. In addition, SP1 was found to upregulate ZFAS1 transcription by binding to its promoter region. ZFAS1 knockdown resulted in a significant reduction of AKT/mTOR pathway components, including p-AKT, AKT, p-mTOR, and mTOR. When the AKT activator SC79 was introduced, the repressive effects of ZFAS1 knockdown on cell invasion, migration, proliferation, and AKT/mTOR signaling were partially reversed.
Our results highlight the pivotal role of ZFAS1 in gastric cancer cell malignancy, which inhibits the activation of the AKT/mTOR pathway. The regulatory involvement of SP1 in ZFAS1 transcription provides a novel understanding of the molecular mechanisms driving cancer progression and offers potential therapeutic avenues by suggesting that further research could focus on developing targeted therapies that modulate ZFAS1 expression or activity, which may lead to more effective treatment options for gastric cancer patients in the future.
本研究旨在探讨长链非编码RNA ZFAS1在胃癌进展中的作用，重点关注其受SP1调控的机制及其对AKT/mTOR信号通路的影响。通过探究ZFAS1对细胞增殖、迁移和凋亡的作用，我们试图揭示其分子机制及作为治疗靶点的潜力。
我们通过逆转录定量聚合酶链反应（RT-qPCR）评估了胃癌细胞（SGC7901）中ZFAS1的表达，并与GES-1细胞进行比较。LnCAR数据库提供了与正常组织相比，STAD中ZFAS1水平的相关信息。为了在SGC7901细胞中敲低ZFAS1，我们用si-ZFAS1 #1-3（靶向ZFAS1的小干扰RNA）转染细胞，发现si-ZFAS1-2具有最高的敲低效率。然后，使用Transwell侵袭实验、伤口愈合实验、CCK8和流式细胞术评估敲低ZFAS1对细胞侵袭、迁移和增殖的影响。此外，使用JASPAR数据库和随后的染色质免疫沉淀实验（ChIP）检测ZFAS1启动子区域，以了解SP1转录因子的结合情况。使用蛋白质免疫印迹法阐明ZFAS1对AKT/mTOR通路的影响。
结果显示SGC7901细胞中ZFAS1表达增加，这与胃癌的不良预后相关。在SGC7901细胞中敲低ZFAS1可抑制细胞侵袭、迁移和增殖，并诱导凋亡。此外，发现SP1通过结合其启动子区域上调ZFAS1转录。敲低ZFAS1导致AKT/mTOR通路成分，包括p-AKT、AKT、p-mTOR和mTOR显著减少。当引入AKT激活剂SC79时，敲低ZFAS1对细胞侵袭、迁移、增殖和AKT/mTOR信号的抑制作用部分被逆转。
我们的结果突出了ZFAS1在胃癌细胞恶性肿瘤中的关键作用，其抑制了AKT/mTOR通路的激活。SP1对ZFAS1转录的调控参与为驱动癌症进展的分子机制提供了新的认识，并通过表明进一步的研究可以集中于开发调节ZFAS1表达或活性的靶向治疗方法，提供了潜在的治疗途径，这可能在未来为胃癌患者带来更有效的治疗选择。
