Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of University of South China, Hengyang, Hunan, P.R. China.
Anticancer Drugs. 2021 Jan 1;32(1):1-10. doi: 10.1097/CAD.0000000000000990.
Circular RNAs are involved in the occurrence and development of different types of cancers. We aimed to illustrate the expression profile and mechanism of circ_0074027 in non-small cell lung cancer (NSCLC). Quantitative real-time PCR was employed to detect the expression of circ_0074027, paired like homeodomain 1 (PITX1) mRNA (mPITX1) and microRNA-362-3p (miR-362-3p). Western blot assay was utilized to measure the levels of clathrin heavy chain (CLTC), cyclin D1, BCL2-associated X, apoptosis regulator Bax (Bax), vimentin and matrix metallopeptidase 9. The clonogenicity, apoptosis and metastasis of NSCLC cells were examined by colony formation assay, flow cytometry and transwell migration and invasion assays. The target relationship between miR-362-3p and circ_0074027 or CLTC was predicted by starBase website and was validated by dual-luciferase reporter assay. Murine xenograft assay was applied to explore the function of circ_0074027 in vivo. We found that The enrichment of circ_0074027 and CLTC protein was elevated, and a significant reduction in the expression of miR-362-3p was observed in NSCLC tissues and cells relative to adjacent normal tissues and human bronchial epithelial cells 16HBE. Circ_0074027 possessed a stable circular structure. Circ_0074027 and CLTC could accelerate the colony formation and metastasis and suppress the apoptosis of NSCLC cells. Circ_0074027/miR-362-3p/CLTC axis was first found to regulate the malignance of NSCLC cells. The biological influence caused by circ_0074027 depletion on NSCLC cells was alleviated by the accumulation of CLTC. Circ_0074027 acted as an oncogene to promote the growth of NSCLC tumors in vivo. In conclusion, Circ_0074027 contributed to the progression of NSCLC through promoting the proliferation and motility while hampering the apoptosis of NSCLC cells via miR-362-3p/CLTC axis.
环状 RNA 参与了多种类型癌症的发生和发展。我们旨在阐述 circ_0074027 在非小细胞肺癌(NSCLC)中的表达谱和作用机制。采用实时定量 PCR 检测 circ_0074027、配对同源域 1 (PITX1) mRNA (mPITX1) 和 microRNA-362-3p (miR-362-3p) 的表达。采用 Western blot 检测笼蛋白重链 (CLTC)、细胞周期蛋白 D1、B 细胞淋巴瘤-2 相关 X 蛋白、凋亡调节因子 Bax (Bax)、波形蛋白和基质金属蛋白酶 9 的水平。通过集落形成实验、流式细胞术和 Transwell 迁移和侵袭实验检测 NSCLC 细胞的克隆形成、凋亡和转移。通过 starBase 网站预测 miR-362-3p 与 circ_0074027 或 CLTC 的靶关系,并通过双荧光素酶报告基因实验进行验证。应用小鼠异种移植实验研究 circ_0074027 在体内的功能。我们发现,与相邻正常组织和人支气管上皮细胞 16HBE 相比,NSCLC 组织和细胞中 circ_0074027 和 CLTC 蛋白的丰度升高,miR-362-3p 的表达显著降低。Circ_0074027 具有稳定的环状结构。Circ_0074027 和 CLTC 可加速 NSCLC 细胞的集落形成和转移,抑制细胞凋亡。首次发现 circ_0074027/miR-362-3p/CLTC 轴调节 NSCLC 细胞的恶性程度。Circ_0074027 耗竭对 NSCLC 细胞的生物学影响可通过 CLTC 的积累而减轻。Circ_0074027 作为一种癌基因,促进体内 NSCLC 肿瘤的生长。总之,Circ_0074027 通过促进增殖和运动,同时通过 miR-362-3p/CLTC 轴抑制 NSCLC 细胞的凋亡,促进 NSCLC 的进展。