Xie Dan, Zhang Shuyi, Jiang Xiaocong, Huang Weizhen, He Ying, Li Yi, Chen Sihan, Xiong Hailin
Department of Medical Oncology, Huizhou Municipal Central Hospital of Guangdong Province, No. 41, North Eling Road, Huizhou, 516000, Guangdong, People's Republic of China.
Department of Radiotherapy Oncology, Huizhou Municipal Central Hospital of Guangdong Province, Huizhou, Guangdong, People's Republic of China.
Biochem Genet. 2023 Feb;61(1):1-20. doi: 10.1007/s10528-022-10231-6. Epub 2022 Jun 9.
In this study, we explored the role of circ_CSPP1 in non-small cell lung cancer (NSCLC) using NSCLC cell lines (A549 and H1299) and human bronchial epithelioid cells (16HBE). The differential expression of circ_CSPP1, miR-486-3p and BRD9 in NSCLC by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot in A549 cells, H1299 cells, 16HBE cells, NSCLC tissues and healthy lung tissues. Dual-luciferase reporter assay was conducted to verify the interaction between circ_CSPP1 and miR-486-3p or miR-486-3p and BRD9. The effect of circ_CSPP1/miR-486-3p/BRD9 axis on NSCLC cell proliferation was evaluated using cell counting kit-8 assay, colony formation assay, and 5-ethynyl-2'-deoxyuridine assay. Additionally, transwell assays were performed to evaluate the effect of circ_CSPP1/miR-486-3p/BRD9 axis on A549 and H1299 cell migration and invasion. The effect of circ_CSPP1 on tumor tumorigenesis of A549 cells in vivo was determined by xenograft tumor model and immunohistochemistry assay. Circ_CSPP1 and BRD9 expression were upregulated, while miR-486-3p expression was downregulated in tumor tissues of NSCCL patients and A549 and H1299 cells. Circ_CSPP1 specifically bound miR-486-3p, and miR-486-3p could target BRD9. Circ_CSPP1 upregulation promoted proliferation, invasion and migration of NSCLC cells, circ_CSPP1 knockdown or miR-486-3p upregulation had the opposite effects. Circ_CSPP1 knockdown-induced effects were reverted by miR-486-3p inhibition. Similarly, the effects of miR-486-3p upregulation on NSCLC cell proliferation, invasion and migration were reversed by BRD9 overexpression. In addition, circ_CSPP1 silencing inhibited tumor growth in nude mice. Circ_CSPP1 promoted A549 and H1299 cell malignancy by competitively inhibiting BRD9 and binding to miR-486-3p.
在本研究中，我们使用非小细胞肺癌（NSCLC）细胞系（A549和H1299）以及人支气管上皮样细胞（16HBE），探讨了circ_CSPP1在非小细胞肺癌中的作用。通过定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测A549细胞、H1299细胞、16HBE细胞、NSCLC组织和健康肺组织中circ_CSPP1、miR-486-3p和BRD9的差异表达。进行双荧光素酶报告基因检测以验证circ_CSPP1与miR-486-3p或miR-486-3p与BRD9之间的相互作用。使用细胞计数试剂盒-8检测、集落形成检测和5-乙炔基-2'-脱氧尿苷检测评估circ_CSPP1/miR-486-3p/BRD9轴对NSCLC细胞增殖的影响。此外，进行Transwell检测以评估circ_CSPP1/miR-486-3p/BRD9轴对A549和H1299细胞迁移和侵袭的影响。通过异种移植肿瘤模型和免疫组织化学检测确定circ_CSPP1对A549细胞体内肿瘤发生的影响。在NSCCL患者的肿瘤组织以及A549和H1299细胞中，circ_CSPP1和BRD9表达上调，而miR-486-3p表达下调。circ_CSPP1特异性结合miR-486-3p，且miR-486-3p可靶向BRD9。circ_CSPP1上调促进NSCLC细胞的增殖、侵袭和迁移，circ_CSPP1敲低或miR-486-3p上调则产生相反的效果。miR-486-3p抑制可逆转circ_CSPP1敲低诱导的效应。同样，BRD9过表达可逆转miR-486-3p上调对NSCLC细胞增殖、侵袭和迁移的影响。此外，circ_CSPP1沉默抑制裸鼠肿瘤生长。circ_CSPP1通过竞争性抑制BRD9并与miR-486-3p结合促进A549和H1299细胞的恶性表型。
