Wang Huimin, Tang Zhiqin, Duan Jihui, Zhou Chunlei, Xu Ke, Mu Hong
Department of Clinical Laboratory, Tianjin First Center Hospital, Tianjin, Hebei, China.
Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenviroment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin, Hebei, China.
Bioengineered. 2022 Apr;13(4):8937-8949. doi: 10.1080/21655979.2022.2056822.
Circular RNA (circRNA) circ_0008717 has been revealed to promote cell carcinogenesis in non-small cell lung cancer (NSCLC). Exosomal circRNA packaged into exosomes has been defined as a potential diagnostic and therapeutic biomarker of cancers. However, little attention is focused on the role of circRNAs within exosomes in NSCLC. Exosomes were isolated by ultracentrifugation method and qualified by nanoparticle tracking analysis and Western blot. Levels of circ_0008717, microRNA (miR)-1287-5p, and P21-activated kinase 2 (PAK2) were detected using qRT-PCR and western blot. The interaction between miR-1287-5p and circ_0008717 or PAK2 was investigated. The phenotypes of NSCLC cells with circ_0008717 downregulation were tested. Circ_0008717 was highly expressed in NSCLC. Functionally, circ_0008717 deficiency suppressed cell malignant phenotypes in NSCLC and in nude mice. Circ_0008717 sponged miR-1287-5p to elevate PAK2, a downstream target of miR-1287-5p. Silencing of miR-1287-5p blocked the antitumor effects of circ_0008717 knockdown in NSCLC cells. Besides, miR-1287-5p repressed cell oncogenic behaviors in NSCLC by targeting PAK2. Besides that, we confirmed that circ_0008717 was incorporated into exosomes in NSCLC cells. Circ_0008717 knockdown inhibited NSCLC tumorigenesis via miR-1287-5p/PAK2 axis, and the extracellular circulating circ_0008717 was transferred through incorporation in exosomes.
环状RNA(circRNA)circ_0008717已被证实可促进非小细胞肺癌(NSCLC)的细胞癌变。包装在外泌体中的外泌体circRNA已被定义为癌症的潜在诊断和治疗生物标志物。然而,外泌体内circRNA在NSCLC中的作用鲜受关注。采用超速离心法分离外泌体,并通过纳米颗粒跟踪分析和蛋白质印迹法进行鉴定。使用qRT-PCR和蛋白质印迹法检测circ_0008717、微小RNA(miR)-1287-5p和p21激活激酶2(PAK2)的水平。研究了miR-1287-5p与circ_0008717或PAK2之间的相互作用。检测了circ_0008717下调的NSCLC细胞的表型。Circ_0008717在NSCLC中高表达。在功能上,circ_0008717缺陷抑制了NSCLC细胞和裸鼠的细胞恶性表型。Circ_0008717吸附miR-1287-5p以提高PAK2(miR-1287-5p的下游靶点)的水平。miR-1287-5p的沉默阻断了circ_0008717敲低对NSCLC细胞的抗肿瘤作用。此外,miR-1287-5p通过靶向PAK2抑制NSCLC中的细胞致癌行为。除此之外,我们证实circ_0008717被纳入NSCLC细胞的外泌体中。Circ_0008717敲低通过miR-1287-5p/PAK2轴抑制NSCLC肿瘤发生,并且细胞外循环的circ_0008717通过纳入外泌体进行转移。