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环状RNA circ_0020123通过吸附miR-590-5p调节THBS2促进非小细胞肺癌进展。

Circular RNA circ_0020123 promotes non-small cell lung cancer progression by sponging miR-590-5p to regulate THBS2.

作者信息

Wang Liang, Zhao Lantao, Wang Yonghong

机构信息

Department of Thoracic Surgery, Lianyungang Second People's Hospital, No. 41 Hailian East Road, Haizhou District, Lianyungang, 222006 Jiangsu China.

出版信息

Cancer Cell Int. 2020 Aug 11;20:387. doi: 10.1186/s12935-020-01444-z. eCollection 2020.

Abstract

BACKGROUND

The incidence and death rate of non-small cell lung cancer (NSCLC) in China ranks the first among the malignant tumors. Circular RNA (circRNA) was reported to be involved in the progression of NSCLC. Our study aimed to investigate the underlying mechanism of circ_0020123 in NSCLC progression.

METHODS

Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of circ_0020123, miR-590-5p and Thrombospondin 2 (THBS2) in NSCLC tissues and cells. Cell proliferation and migration were examined by Cell Counting Kit-8 (CCK-8) assay and Transwell assay, respectively. Flow cytometry assay was used to detect the apoptosis of NSCLC cells. The protein levels of Ki-67, matrix metalloprotein-9 (MMP-9), Cleaved-caspase9 (Cleaved-casp9) and THBS2 were detected by Western blot. The targets of circ_0020123 and miR-590-5p were predicted by starBase 3.0 and TargetScan, and then confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. The animal experiment showed the effect of circ_0020123 on tumor growth in vivo.

RESULTS

The expression of circ_0020123 was upregulated in NSCLC tissues and cells. Functionally, circ_0020123 downregulation inhibited the proliferation and migration and promoted the apoptosis of NSCLC cells. Interestingly, circ_0020123 directly targeted miR-590-5p, and inhibition of miR-590-5p reversed the knockdown effects of circ_0020123 on NSCLC cells. More importantly, THBS2 was a target of miR-590-5p, and THBS2 overexpression reversed the effects of circ_0020123 knockdown on cell proliferation, migration and apoptosis in NSCLC cells. Finally, suppression of circ_0020123 inhibited tumor growth in vivo through miR-590-5p/THBS2 axis.

CONCLUSION

Circular RNA circ_0020123 regulated THBS2 by sponging miR-590-5p to promote cell proliferation and migration and inhibit cell apoptosis in NSCLC cells.

摘要

背景

中国非小细胞肺癌(NSCLC)的发病率和死亡率在恶性肿瘤中位居首位。据报道,环状RNA(circRNA)参与了NSCLC的进展。本研究旨在探讨circ_0020123在NSCLC进展中的潜在机制。

方法

采用定量实时聚合酶链反应(qRT-PCR)检测circ_0020123、miR-590-5p和血小板反应蛋白2(THBS2)在NSCLC组织和细胞中的表达。分别通过细胞计数试剂盒-8(CCK-8)检测法和Transwell检测法检测细胞增殖和迁移。采用流式细胞术检测NSCLC细胞的凋亡。通过蛋白质免疫印迹法检测Ki-67、基质金属蛋白酶-9(MMP-9)、裂解的半胱天冬酶9(Cleaved-casp9)和THBS2的蛋白水平。通过starBase 3.0和TargetScan预测circ_0020123和miR-590-5p的靶标,然后通过双荧光素酶报告基因检测法和RNA免疫沉淀(RIP)检测法进行验证。动物实验显示了circ_0020123对体内肿瘤生长的影响。

结果

circ_0020123在NSCLC组织和细胞中的表达上调。在功能上,circ_0020123下调抑制了NSCLC细胞的增殖和迁移,并促进了其凋亡。有趣的是,circ_0020123直接靶向miR-590-5p,抑制miR-590-5p可逆转circ_0020123对NSCLC细胞的敲低作用。更重要的是,THBS2是miR-590-5p的靶标,THBS2过表达可逆转circ_0020123敲低对NSCLC细胞增殖、迁移和凋亡的影响。最后,抑制circ_0020123通过miR-590-5p/THBS2轴抑制体内肿瘤生长。

结论

环状RNA circ_0020123通过海绵吸附miR-590-5p来调节THBS2,从而促进NSCLC细胞的增殖和迁移并抑制其凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0d98/7422555/e29e5156fb5b/12935_2020_1444_Fig1_HTML.jpg

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