环状 RNA RACGAP1 通过 miR-1296/CDK2 通路促进非小细胞肺癌细胞的增殖。

circRACGAP1 Promotes Proliferation of Non-Small Cell Lung Cancer Cells through the miR-1296/CDK2 Pathway.

机构信息

The Affiliated Changsha Central Hospital, Department of Oncology, Hengyang Medical School, University of South China, Changsha, Hunan, China.

出版信息

Folia Biol (Praha). 2024;70(2):104-112. doi: 10.14712/fb2024070020104.

DOI:10.14712/fb2024070020104

PMID:39231318

Abstract

Circular RNAs (circRNAs) have played an essential role in cancer development. This study aimed to illustrate the impact and potential mechanism of circRACGAP1 action in NSCLC development. The expression patterns of circRACGAP1, miR-1296, and CDK2 in NSCLC tissues and cell lines were analysed by RT-qPCR. The function of circRACGAP1 in NSCLC cell proliferation and apoptosis was investigated using the CCK-8 assay, flow cytometry, TUNEL staining, and Western blot. The interaction among circRACGAP1, miR-1296, and CDK2 was clarified by dual-luciferase reporter assay while the correlation was confirmed by the Pearson correlation coefficient. The expression of circRACGAP1 and CDK2 was up-regulated in NSCLC tissues, while the expression of miR-1296 was down-regulated. Cell function studies further revealed that circRACGAP1 could promote NSCLC cell proliferation, accelerate the cell cycle process, up-regulate B-cell lymphoma 2 (Bcl2) expression, and down-regulate Bcl2-associated X (Bax) expression. miR-1296 was identified as a downstream target to reverse circRACGAP1-mediated cell proliferation. miR-1296 directly targeted the 3'-UTR of CDK2 to regulate proliferation and apoptosis of NSCLC cells. Additionally, the dual-luciferase reporter assay and Pearson correlation coefficient analysis proved that circRACGAP1 acted in NSCLC cells by negatively regulating miR-1296 expression and positively regulating CDK2 expression. In summary, our study revealed that circRACGAP1 promoted NSCLC cell proliferation by regulating the miR-1296/CDK2 pathway, providing potential diagnostic and therapeutic targets for NSCLC.

摘要

环状 RNA（circRNAs）在癌症发展中发挥了重要作用。本研究旨在阐明 circRACGAP1 在非小细胞肺癌（NSCLC）发展中的作用及其潜在机制。通过 RT-qPCR 分析 NSCLC 组织和细胞系中 circRACGAP1、miR-1296 和 CDK2 的表达模式。通过 CCK-8 测定、流式细胞术、TUNEL 染色和 Western blot 研究 circRACGAP1 在 NSCLC 细胞增殖和凋亡中的作用。通过双荧光素酶报告基因实验阐明 circRACGAP1、miR-1296 和 CDK2 之间的相互作用，通过 Pearson 相关系数证实它们之间的相关性。circRACGAP1 和 CDK2 的表达在 NSCLC 组织中上调，而 miR-1296 的表达下调。细胞功能研究进一步表明，circRACGAP1 可促进 NSCLC 细胞增殖，加速细胞周期进程，上调 B 细胞淋巴瘤 2（Bcl2）表达，下调 Bcl2 相关 X（Bax）表达。miR-1296 被鉴定为逆转 circRACGAP1 介导的细胞增殖的下游靶标。miR-1296 直接靶向 CDK2 的 3'-UTR 来调节 NSCLC 细胞的增殖和凋亡。此外，双荧光素酶报告基因实验和 Pearson 相关系数分析证明，circRACGAP1 通过负调控 miR-1296 表达和正调控 CDK2 表达在 NSCLC 细胞中发挥作用。总之，我们的研究表明，circRACGAP1 通过调节 miR-1296/CDK2 通路促进 NSCLC 细胞增殖，为 NSCLC 提供了潜在的诊断和治疗靶点。