Gong Lei, Chen Juan, Jiang Xijiao
Department of Otolaryngology, Affiliated Hospital of Shandong Medical College, Qingdao.
Department of Otolaryngology, The First People's Hospital of Jiangxia District of Wuhan, Union Jiangnan Hospital, Wuhan, China.
Anticancer Drugs. 2022 Mar 1;33(3):245-256. doi: 10.1097/CAD.0000000000001260.
Transcriptome expression profiles of laryngeal squamous cell carcinoma (LSCC) are altered, and we aimed to investigate expression and role of hsa_circ_0005033 (circ_0005033), microRNA (miR)-107 and insulin-like growth factor 1 receptor (IGF1R) in LSCC. Real-time PCR, western blotting and immunohistochemistry detected RNA and protein expression levels. Functional assays were performed using MTT assay, EdU assay, apoptosis assay, flow cytometry, Transwell assay, and xenograft tumor model. Direct interaction was predicted by Starbase algorithm and validated by dual-luciferase reporter assay and RNA immunoprecipitation. Expression of circ_0005033 was substantially upregulated in LSCC tissues and cells, and allied with miR-107 downregulation and IGF1R upregulation. Circ_0005033 showed a closed-loop structure and long half-life. Essentially, circ_0005033 and IGF1R were competing endogenous RNAs for miR-107 via target binding. Silencing circ_0005033 facilitated apoptosis rate and lowered cell viability, proliferation, migration and invasion of LSCC cells, as well as delayed xenograft tumor growth. Allied with that, cleaved-caspase 3/8/9 expression was elevated via death receptor-mediated and mitochondrial pathways, and expression of matrix metalloproteinase-2 (MMP2), MMP9, cyclin D1 and proliferating cell nuclear antigen was decreased. Moreover, Cisplatin-induced inhibition of cell viability was exacerbated by inhibiting circ_0005033. These functional effects of circ_0005033 depression were consistent with those of miR-107 overexpression. Furthermore, depleting miR-107 and restoring IGF1R abated the effects of circ_0005033 knockdown and miR-107 overexpression, respectively. Circ_0005033 was oncogenic in LSCC by regulating cell progression and Cisplatin sensitivity at least via miR-107/IGF1R axis.
喉鳞状细胞癌（LSCC）的转录组表达谱发生改变，我们旨在研究hsa_circ_0005033（circ_0005033）、微小RNA（miR）-107和胰岛素样生长因子1受体（IGF1R）在LSCC中的表达及作用。采用实时定量PCR、蛋白质免疫印迹法和免疫组织化学检测RNA和蛋白质表达水平。使用MTT法、EdU法、凋亡检测、流式细胞术、Transwell法和异种移植肿瘤模型进行功能分析。通过Starbase算法预测直接相互作用，并通过双荧光素酶报告基因检测和RNA免疫沉淀进行验证。circ_0005033在LSCC组织和细胞中表达显著上调，与miR-107下调和IGF1R上调相关。circ_0005033呈闭环结构且半衰期长。本质上，circ_0005033和IGF1R通过靶标结合作为miR-107的竞争性内源性RNA。沉默circ_0005033可促进LSCC细胞的凋亡率，降低细胞活力、增殖、迁移和侵袭能力，并延缓异种移植肿瘤生长。与此同时，通过死亡受体介导的途径和线粒体途径，裂解的半胱天冬酶3/8/9表达升高，基质金属蛋白酶-2（MMP2）、MMP9、细胞周期蛋白D1和增殖细胞核抗原的表达降低。此外，抑制circ_0005033可加剧顺铂诱导的细胞活力抑制。circ_0005033抑制的这些功能效应与miR-107过表达的效应一致。此外，分别消耗miR-107和恢复IGF1R可减轻circ_0005033敲低和miR-107过表达的效应。circ_0005033至少通过miR-107/IGF1R轴调节细胞进程和顺铂敏感性，在LSCC中具有致癌作用。
