Zhang Hui, Cai Wenwen, Miao Yiyan, Gu Yihang, Zhou Xiaorong, Kaneda Hiroyasu, Wang Lan
The Jiangyin Clinical College of Xuzhou Medical University, Xuzhou, China.
Sanmen County People's Hospital, Taizhou, China.
J Cell Mol Med. 2024 Dec;28(23):e70270. doi: 10.1111/jcmm.70270.
Long non-coding RNA (lncRNA) LINC01116 is crucial in promoting cell proliferation, invasion and migration in solid tumours, including lung adenocarcinoma (LUAD). LINC01116 acts as a competing endogenous RNAs (ceRNA) that binds competitively to microRNAs and plays a critical role in tumour migration and invasion. However, other mechanisms of action besides the ceRNA theory have been rarely reported and remain to be elucidated further. The differences in RNA and protein levels in cells and tissues were assessed through real-time quantitative PCR and Western blot analysis. In vitro functional assays and in vivo xenograft models were used to analyse the function of LINC01116 in LUAD. Thus, the molecular correlation between miR-9-5p and CCNE1 was investigated through direct and indirect mechanism experiments. LINC01116, miR-9-5p and CCNE1 were upregulated in LUAD cell lines and tissues and were associated with a poor prognosis in patients. LINC01116 depletion inhibited proliferation but facilitated cell apoptosis. AGO2-RNA binding protein immunoprecipitation (AGO2-RIP) experiments confirmed that AGO2 binds to LINC01116 and miR-9-5p, indicating that LINC01116 interacts with miR-9-5p. The overexpression of miR-9-5p and CCNE1 effectively counteracts the biological effects of LINC01116 knockdown on reduced proliferation and cell cycle arrest in LUAD cells. The downregulation of miR-9-5p significantly reduces the CCNE1 level in A549 cells, and the upregulation of LINC01116 counteracts the downregulation of miR-9-5p effect, restoring the expression level of CCNE1. Our data demonstrated that LINC01116 regulates the expression of CCNE1 by positively regulating miR-9-5p, thereby affecting cell cycle, proliferation and participating in the development of LUAD.
长链非编码RNA（lncRNA）LINC01116在促进实体瘤（包括肺腺癌，LUAD）的细胞增殖、侵袭和迁移中起关键作用。LINC01116作为一种竞争性内源性RNA（ceRNA），与微小RNA竞争性结合，并在肿瘤迁移和侵袭中起关键作用。然而，除ceRNA理论外的其他作用机制鲜有报道，仍有待进一步阐明。通过实时定量PCR和蛋白质免疫印迹分析评估细胞和组织中RNA和蛋白质水平的差异。采用体外功能测定和体内异种移植模型分析LINC01116在LUAD中的功能。因此，通过直接和间接机制实验研究了miR-9-5p与CCNE1之间的分子相关性。LINC01116、miR-9-5p和CCNE1在LUAD细胞系和组织中上调，且与患者预后不良相关。LINC01116的缺失抑制增殖，但促进细胞凋亡。AGO2-RNA结合蛋白免疫沉淀（AGO2-RIP）实验证实AGO2与LINC01116和miR-9-5p结合，表明LINC01116与miR-9-5p相互作用。miR-9-5p和CCNE1的过表达有效抵消了LINC01116敲低对LUAD细胞增殖减少和细胞周期停滞的生物学效应。miR-9-5p的下调显著降低A549细胞中CCNE1的水平，而LINC01116的上调抵消了miR-9-5p下调的作用，恢复了CCNE1的表达水平。我们的数据表明，LINC01116通过正向调节miR-9-5p来调节CCNE1的表达，从而影响细胞周期、增殖并参与LUAD的发生发展。
