Department of Biochemistry and Molecular Biology, Binzhou Medical University, Yantai, China.
Yantai Affiliated Hospital of Binzhou Medical University, Yantai, China.
Thorac Cancer. 2024 Sep;15(27):1929-1945. doi: 10.1111/1759-7714.15396. Epub 2024 Aug 7.
The aim of the present study was to investigate the function of novel circular RNA hsa_circ_0036683 (circ-36683) in non-small cell lung cancer (NSCLC).
RNA sequencing was used to screen out differentially expressed miRNAs. Expression levels of miR-4664-3p and circ-36683 were evaluated in lung carcinoma cells and tissues by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The effects of miR-4664-3p and circ-36683 on proliferation and migration were assessed using cell counting kit-8 (CCK-8), wound healing and transwell migration assays and xenograft experiments. The targeting relationship of circ-36683/miR-4664-3p/CDK2AP2 was assessed by luciferase reporter assays, western blot, qRT-PCR and argonaute2-RNA immunoprecipitation (AGO2 RIP). Co-immunoprecipitation (Co-IP), 5-ethynyl-2'-deoxyuridine (EdU) staining and CCK-8 were used to validate the indispensable role of CDK2AP2 in suppressing cell proliferation as a result of CDK2AP1 overexpression.
By RNA sequencing, miR-4664-3p was screened out as an abnormally elevated miRNA in NSCLC tissues. Transfection of miR-4664-3p could promote cell proliferation, migration and xenograft tumor growth. As a target of miR-4664-3p, CDK2AP2 expression was downregulated by miR-4664-3p transfection and CDK2AP2 overexpression could abolish the proliferation promotion resulting from miR-4664-3p elevation. Circ-36683, derived from back splicing of ABHD2 pre-mRNA, was attenuated in NSCLC tissue and identified as a sponge of miR-4664-3p. The functional study revealed that circ-36683 overexpression suppressed cell proliferation, migration and resulted in G0/G1 phase arrest. More importantly, the antioncogenic function of circ-36683 was largely dependent on the miR-4664-3p/CDK2AP2 axis, through which circ-36683 could upregulate the expression of p53/p21/p27 and downregulate the expression of CDK2/cyclin E1.
The present study revealed the antioncogenic role of circ-36683 in suppressing cell proliferation and migration and highlighted that targeting the circ-36683/miR-4664-3p/CDK2AP2 axis is a promising strategy for the intervention of NSCLC.
本研究旨在探究新型环状 RNA hsa_circ_0036683（circ-36683）在非小细胞肺癌（NSCLC）中的作用。
采用 RNA 测序筛选差异表达的 miRNAs。采用实时定量逆转录聚合酶链反应（qRT-PCR）检测肺癌细胞和组织中 miR-4664-3p 和 circ-36683 的表达水平。通过细胞计数试剂盒-8（CCK-8）、划痕愈合和 Transwell 迁移实验以及异种移植实验评估 miR-4664-3p 和 circ-36683 对增殖和迁移的影响。通过荧光素酶报告基因实验、Western blot、qRT-PCR 和 Argonaute2-RNA 免疫沉淀（AGO2 RIP）评估 circ-36683/miR-4664-3p/CDK2AP2 的靶向关系。采用 Co-immunoprecipitation（Co-IP）、5-乙炔基-2'-脱氧尿苷（EdU）染色和 CCK-8 验证 CDK2AP1 过表达抑制细胞增殖的不可或缺作用是由于 CDK2AP2 的抑制。
通过 RNA 测序，筛选出 miR-4664-3p 作为 NSCLC 组织中异常上调的 miRNA。miR-4664-3p 的转染可促进细胞增殖、迁移和异种移植肿瘤生长。作为 miR-4664-3p 的靶标，CDK2AP2 的表达被 miR-4664-3p 转染下调，CDK2AP2 的过表达可消除 miR-4664-3p 升高导致的增殖促进作用。Circ-36683 来源于 ABHD2 前体 mRNA 的反向剪接，在 NSCLC 组织中减少，并被鉴定为 miR-4664-3p 的海绵。功能研究表明，circ-36683 过表达抑制细胞增殖、迁移并导致 G0/G1 期阻滞。更重要的是，circ-36683 的抑癌作用在很大程度上依赖于 miR-4664-3p/CDK2AP2 轴，通过该轴，circ-36683 可上调 p53/p21/p27 的表达并下调 CDK2/细胞周期蛋白 E1 的表达。
本研究揭示了 circ-36683 在抑制细胞增殖和迁移中的抑癌作用，并强调靶向 circ-36683/miR-4664-3p/CDK2AP2 轴是 NSCLC 干预的一种有前途的策略。
