Yang Wenhui, Wu Lina, Jin Mingming
Department of Internal Medicine, Shanghai Ruici Ruijie Outpatient Department, Yangpu District, Shanghai, China.
Department of General Practice, Renji Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, China.
J Clin Lab Anal. 2022 Apr;36(4):e24262. doi: 10.1002/jcla.24262. Epub 2022 Feb 25.
Circular RNAs hold significant regulatory functions during various tumors. However, the exact hsa_circ_0041268 roles in non-small cell lung cancer (NSCLC) along with regulatory mechanism are unknown. In this study, RT-qPCR was used to perceive hsa_circ_0041268 expressions in NSCLC cell lines. Our team constructed small interfering RNA for hsa_circ_0041268. NSCLC cell proliferation, migration, and tumorigenesis in nude mice were assayed to confirm hsa_circ_0041268 activities in NSCLC cells. We then used bioinformatics and luciferase reporter analyses to characterize the hsa_circ_0041268 downstream targets. The result shows that the expressions of hsa_circ_0041268 incremented in NSCLC cell lines and hsa_circ_0041268 downregulation decreased cell proliferation and migration. ROCK1 and miR-214-5p were hsa_circ_0041268 downstream targets. miR-214-5p downregulation or ROCK1 overexpression restored migration and proliferation abilities after hsa_circ_0041268 silencing. ROCK1 overexpression renovated migration and proliferation abilities after miR-214-5p overexpression. In vivo investigations confirmed that hsa_circ_0041268 downregulation inhibited tumor formation and metastasis in nude mice xenografts. Together, results demonstrated that hsa_circ_0041268 acted as tumor promoter through novel hsa_circ_0041268/miR-214-5p/ROCK1 axis, which highlighted its potential as NSCLC therapeutic agent.
环状RNA在多种肿瘤中发挥着重要的调节功能。然而，hsa_circ_0041268在非小细胞肺癌（NSCLC）中的具体作用及其调控机制尚不清楚。在本研究中，采用RT-qPCR检测NSCLC细胞系中hsa_circ_0041268的表达。我们的团队构建了针对hsa_circ_0041268的小干扰RNA。通过检测NSCLC细胞增殖、迁移以及裸鼠体内肿瘤发生情况，以证实hsa_circ_0041268在NSCLC细胞中的活性。随后，我们利用生物信息学和荧光素酶报告基因分析来鉴定hsa_circ_0041268的下游靶点。结果显示，hsa_circ_0041268在NSCLC细胞系中的表达增加，下调hsa_circ_0041268可降低细胞增殖和迁移能力。ROCK1和miR-214-5p是hsa_circ_0041268的下游靶点。沉默hsa_circ_0041268后，下调miR-214-5p或过表达ROCK1可恢复细胞的迁移和增殖能力。过表达miR-214-5p后，过表达ROCK1可恢复细胞的迁移和增殖能力。体内研究证实，下调hsa_circ_0041268可抑制裸鼠异种移植瘤的肿瘤形成和转移。综上所述，结果表明hsa_circ_0041268通过新的hsa_circ_0041268/miR-214-5p/ROCK1轴发挥肿瘤促进作用，这突出了其作为NSCLC治疗药物的潜力。
