Li Wen, Wang Shanshan, Shan Boquan, Cheng Xiang, He Hui, Qin Jianbing, Tang Yi, Zhao Heyan, Tian Meiling, Zhang Xinhua, Jin Guohua
Department of Anatomy and Neurobiology, Jiangsu Key Laboratory of Neuroregeneration, Collaborative Innovation Center of Neuroregeneration, Nantong University, Nantong, China.
Front Oncol. 2021 May 17;11:666391. doi: 10.3389/fonc.2021.666391. eCollection 2021.
Glioblastoma multiform (GBM) is the most common and malignant primary brain cancer in adults, and thus, novel potential therapeutic targets for diagnosis and treatment are urgently needed. Circular RNAs (circRNAs) are a class of widespread and diverse endogenous RNAs that have been suggested as potential critical mediators during progression of various tumors. In this study, we investigated the involvement of circHECTD1 in GBM progression. CircHECTD1 Lentivirus, miR-320-5p mimic, and SLC2A1 Lentivirus were transduced into cancer cells independently or together. circHECTD1, miR-320-5p, and SLC2A1 level were detected by qRT-PCR. Western blot and qRT-PCR were applied to measure the expression of SLC2A1, CyclinD1, CDK2, and PCNA. Flow cytometry, EdU, colony formation, Transwell and wound-healing assays were conducted to assess cell proliferation and migration. Luciferase reporter assays were performed to determine the effect of miR-320-5p on circHECTD1 or SLC2A1. Xenograft experiments were implemented to evaluate tumor growth . CircHECTD1 expression led to the promotion of proliferation and migration of GBM cells. In addition, circHECTD1 acted as a ceRNA to interact with miR-320-5p, which targeted the solute carrier family 2 member 1 (SLC2A1). In vivo experiments also revealed that circHECTD1 promoted tumor growth. Collectively, our findings showed that the circHECTD1-miR-320-5p-SLC2A1 regulatory pathway promoted the progression of GBM, suggesting that circHECTD1 may be a therapeutic target for GBM.
多形性胶质母细胞瘤(GBM)是成人中最常见且恶性程度最高的原发性脑癌,因此,迫切需要用于诊断和治疗的新型潜在治疗靶点。环状RNA(circRNAs)是一类广泛且多样的内源性RNA,已被认为是各种肿瘤进展过程中的潜在关键调节因子。在本研究中,我们调查了circHECTD1在GBM进展中的作用。将circHECTD1慢病毒、miR-320-5p模拟物和SLC2A1慢病毒分别或一起转导到癌细胞中。通过qRT-PCR检测circHECTD1、miR-320-5p和SLC2A1水平。应用蛋白质免疫印迹法和qRT-PCR检测SLC2A1、细胞周期蛋白D1、细胞周期蛋白依赖性激酶2(CDK2)和增殖细胞核抗原(PCNA)的表达。进行流式细胞术、EdU检测、集落形成实验、Transwell实验和伤口愈合实验以评估细胞增殖和迁移。进行荧光素酶报告基因实验以确定miR-320-5p对circHECTD1或SLC2A1的影响。实施异种移植实验以评估肿瘤生长。CircHECTD1表达促进了GBM细胞的增殖和迁移。此外,circHECTD1作为竞争性内源RNA(ceRNA)与miR-320-5p相互作用,miR-320-5p靶向溶质载体家族2成员1(SLC2A1)。体内实验还表明circHECTD1促进肿瘤生长。总体而言,我们的研究结果表明circHECTD1-miR-320-5p-SLC2A1调节通路促进了GBM的进展,提示circHECTD1可能是GBM的一个治疗靶点。