Circ_0000735通过调控miR-635/FAM83F轴增强非小细胞肺癌的增殖、转移和糖酵解。
Circ_0000735 enhances the proliferation, metastasis and glycolysis of non-small cell lung cancer by regulating the miR-635/FAM83F axis.
作者信息
Tai Guigang, Zhang Miao, Liu Fang
机构信息
Department of Emergency, Jiaozhou Central Hospital, Qingdao, Shandong Province, China.
Department of Respiration, Jiaozhou Central Hospital, Qingdao, Shandong Province, China.
出版信息
Exp Lung Res. 2021 Apr;47(3):136-148. doi: 10.1080/01902148.2021.1881188. Epub 2021 Feb 9.
BACKGROUND
Circular RNA (circRNA) is considered to be an important regulator of cancer malignant progression, including non-small cell lung cancer (NSCLC). Circ_0000735 has been found to be associated with NSCLC progression. Therefore, its role and molecular mechanism in NSCLC deserve further exploration.
METHODS
Quantitative real-time PCR (qRT-PCR) was used to measure the expression of circ_0000735, microRNA (miR)-635 and family with sequence similarity 83 member F (FAM83F). Cell proliferation, migration, invasion and apoptosis were determined using cell counting kit 8 assay, colony formation assay, transwell assay and flow cytometry. Cell glycolysis were measured by detecting the glucose consumption and lactate production of cells. Western blot analysis was utilized to test the protein levels of glycolysis markers and FAM83F. The relationship between circ_0000735 and miR-635 or miR-635 and FAM83F was verified by dual-luciferase reporter assay. The effect of circ_0000735 on NSCLC tumor growth was evaluated by constructing xenograft models.
RESULTS
Circ_0000735 was a highly expressed circRNA in NSCLC. Silenced circ_0000735 could inhibit NSCLC cell proliferation, migration, invasion, glycolysis, and increase apoptosis. MiR-635 could be sponged by circ_0000735, and its inhibitor could reverse the regulation of circ_0000735 silencing on NSCLC progression. Moreover, FAM83F was a target of miR-635, and circ_0000735 positively regulated FAM83F by sponging miR-635. The inhibitory effect of miR-635 on NSCLC progression could also be reversed by FAM83F overexpression. Additionally, circ_0000735 knockdown reduced NSCLC tumor growth through regulating miR-635/FAM83F axis.
CONCLUSION
Circ_0000735 promoted NSCLC progression by the miR-635/FAM83F axis, showing that circ_0000735 might be a promising biomarker for NSCLC. Highlights: Circ_0000735 knockdown represses NSCLC cell progression and tumor growth. Circ_0000735 functions as a miR-635 sponge. FAM83F is targeted by miR-635.
背景
环状RNA（circRNA）被认为是癌症恶性进展的重要调节因子，包括非小细胞肺癌（NSCLC）。已发现Circ_0000735与NSCLC进展相关。因此，其在NSCLC中的作用和分子机制值得进一步探索。
方法
采用定量实时聚合酶链反应（qRT-PCR）检测Circ_0000735、微小RNA（miR）-635和序列相似性家族83成员F（FAM83F）的表达。使用细胞计数试剂盒8检测、集落形成试验、Transwell试验和流式细胞术测定细胞增殖、迁移、侵袭和凋亡。通过检测细胞葡萄糖消耗和乳酸生成来测量细胞糖酵解。利用蛋白质印迹分析检测糖酵解标志物和FAM83F的蛋白质水平。通过双荧光素酶报告基因试验验证Circ_0000735与miR-635或miR-635与FAM83F之间的关系。通过构建异种移植模型评估Circ_0000735对NSCLC肿瘤生长的影响。
结果
Circ_0000735是NSCLC中高表达的circRNA。沉默Circ_0000735可抑制NSCLC细胞增殖、迁移、侵袭、糖酵解，并增加细胞凋亡。miR-635可被Circ_0000735吸附，其抑制剂可逆转Circ_0000735沉默对NSCLC进展的调节作用。此外,FAM83F是miR-635的靶标，Circ_0000735通过吸附miR-635正向调节FAM83F。FAM83F过表达也可逆转miR-635对NSCLC进展的抑制作用。此外，Circ_0000735敲低通过调节miR-635/FAM83F轴减少NSCLC肿瘤生长。
结论
Circ_0000735通过miR-635/FAM83F轴促进NSCLC进展，表明Circ_0000735可能是NSCLC有前景的生物标志物。要点：Circ_0000735敲低抑制NSCLC细胞进展和肿瘤生长。Circ_0000735作为miR-635的吸附分子发挥作用。FAM83F是miR-635的靶标。
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