Ma Qingshan, Huai Baogang, Liu Yuting, Jia Zhongyao, Zhao Qilong
Department of Oncology, Linyi People's Hospital, Linyi, Shandong 276000, People's Republic of China.
Department of Pulmonary Disease, Pinyi County Hospital of Traditional Chinese Medicine, Linyi, Shandong 273300, People's Republic of China.
Cancer Manag Res. 2021 Jan 7;13:75-87. doi: 10.2147/CMAR.S278913. eCollection 2021.
It was reported that circular RNAs (circRNAs) and microRNAs (miRNAs) were related to non-small cell lung cancer (NSCLC) development. However, the detailed mechanisms of circ_0020123 and miR-384 in NSCLC are elusive.
QRT-PCR and Western blot assay were performed to detect the transcription and protein levels of genes, respectively. Then, the functional experiments, including MTT assay, flow cytometry, and transwell assay, were employed. Besides, the interaction between miR-384 and circ_0020123 or tripartite motif‑containing protein 44 () was predicted by starbase or targetscan, and then verified by the dual-luciferase reporter, RNA pull-down assays and RNA immunoprecipitation assay (RIP). Mouse xenograft assay was performed to evaluate the effect of circ_0020123 on tumor growth in vivo.
Levels of circ_0020123 and were enhanced, and the miR-384 level was attenuated in NSCLC tissues and cells. Circ_0020123 depletion attenuated the abilities of NSCLC cell viability, migration, invasion, and epithelial-mesenchymal transition (EMT), and induced apoptosis. Besides, circ_0020123 interacted with miR-384, and miR-384 targeted . Circ_0020123 regulated cell progression by regulating miR-384 and subsequently mediated expression. Besides, circ_0020123 depletion repressed tumor growth in vivo.
We demonstrated that circ_0020123 knockdown suppressed NSCLC cell progression by regulating the miR-384/ axis, providing the theoretical basis for the therapy of NSCLC.
据报道,环状RNA(circRNAs)和微小RNA(miRNAs)与非小细胞肺癌(NSCLC)的发生发展有关。然而,circ_0020123和miR-384在NSCLC中的具体机制尚不清楚。
分别采用qRT-PCR和蛋白质免疫印迹法检测基因的转录水平和蛋白质水平。然后,进行包括MTT法、流式细胞术和Transwell实验在内的功能实验。此外,通过starbase或targetscan预测miR-384与circ_0020123或含三联基序蛋白44()之间的相互作用,随后通过双荧光素酶报告基因实验、RNA下拉实验和RNA免疫沉淀实验(RIP)进行验证。进行小鼠异种移植实验以评估circ_0020123对体内肿瘤生长的影响。
circ_0020123和的水平在NSCLC组织和细胞中升高,而miR-384水平降低。circ_0020123的缺失减弱了NSCLC细胞的活力、迁移、侵袭及上皮-间质转化(EMT)能力,并诱导细胞凋亡。此外,circ_0020123与miR-384相互作用,且miR-384靶向。circ_0020123通过调控miR-384进而介导的表达来调节细胞进程。此外,circ_0020123的缺失在体内抑制肿瘤生长。
我们证明了circ_0020123的敲低通过调控miR-384/轴抑制NSCLC细胞进程,为NSCLC的治疗提供了理论依据。