Wang Lishui, Xie Yan, Wang Jing, Zhang Ying, Liu Shibiao, Zhan Yao, Zhao Yinghui, Li Juan, Li Peilong, Wang Chuanxin
Department of Clinical Laboratory, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.
Department of Clinical Laboratory, The Second Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.
Front Cell Dev Biol. 2022 May 13;10:833579. doi: 10.3389/fcell.2022.833579. eCollection 2022.
Accumulating literatures suggested that long non-coding RNAs (lncRNAs) were involved in tumorigenesis and cancer progression in lung adenocarcinoma (LUAD). However, the precise regulatory mechanism of lncRNA Lung cancer-associated transcript 1 (LUCAT1) in LUAD is not well defined. In this study, we aimed to investigate the biological function and mechanism of lncRNA LUCAT1 in regulating tumor migration and glycolysis of LUAD. High throughput sequencing was performed to identify differentially expressed lncRNAs between LUAD patients and healthy controls. The expression levels of LUCAT1 in LUAD clinical specimens or cell lines were evaluated by hybridization (ISH) and quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). Functional experiments, including wound-healing, transwell invasion assays, glucose absorption, lactate metabolism and tumor xenograft experiments were conducted to identify the biological functions of LUCAT1 in LUAD. Silencing of LUCAT1, over-expression of LUCAT1 and miR-4316 were generated in LUAD cell lines to verify the regulatory mode of LUCAT1-mir-4316-VEGFA axis. Our findings revealed that lncRNA LUCAT1 was significantly up-regulated in LUAD serum exosomes, tumor tissues, and LUAD cells in comparison with corresponding controls. Receiver operating characteristic curve (ROC) analysis indicated that the area under the curve (AUC) value of serum exosomal LUCAT1 reached 0.852 in distinguishing LUAD patients from healthy individuals. High expression of LUCAT1 in LUAD patient tissues was associated with enhanced Lymph Node Metastasis (LNM), advanced Tumor Node Metastasis (TNM) stage and poorer clinical outcome in LUAD patients. Knockdown of LUCAT1 inhibited LUAD cell metastasis and glycolysis as well as tumor metastasis , while overexpression of LUCAT1 induced a promoted LUAD metastasis and glycolysis. Furthermore, mechanistic investigations revealed that LUCAT1 elevated LUAD cell metastasis and glycolysis by sponging miR-4316, which further led to the upregulation of VEGFA. Finally, the regulatory axis LUCAT1-miR-4316-VEGFA was verified in LUAD. Our present research suggested that LUCAT1 facilitate LUAD cell metastasis and glycolysis serving as a competing endogenous RNA to regulate miR-4316/VEGFA axis, which provided a novel diagnostic marker and therapeutic target for LUAD patients.
越来越多的文献表明，长链非编码RNA（lncRNAs）参与了肺腺癌（LUAD）的肿瘤发生和癌症进展。然而，lncRNA肺癌相关转录本1（LUCAT1）在LUAD中的精确调控机制尚不清楚。在本研究中，我们旨在探讨lncRNA LUCAT1在调节LUAD肿瘤迁移和糖酵解中的生物学功能及机制。进行高通量测序以鉴定LUAD患者和健康对照之间差异表达的lncRNAs。通过杂交（ISH）和定量实时聚合酶链反应（qRT-PCR）评估LUCAT1在LUAD临床标本或细胞系中的表达水平。进行了包括伤口愈合、Transwell侵袭试验、葡萄糖吸收、乳酸代谢和肿瘤异种移植实验在内的功能实验，以确定LUCAT1在LUAD中的生物学功能。在LUAD细胞系中进行LUCAT1沉默、LUCAT1过表达和miR-4316过表达，以验证LUCAT1-miR-4316-VEGFA轴的调控模式。我们的研究结果显示，与相应对照相比，lncRNA LUCAT1在LUAD血清外泌体、肿瘤组织和LUAD细胞中显著上调。受试者工作特征曲线（ROC）分析表明，血清外泌体LUCAT1在区分LUAD患者和健康个体时的曲线下面积（AUC）值达到0.852。LUAD患者组织中LUCAT1的高表达与LUAD患者淋巴结转移（LNM）增加、肿瘤淋巴结转移（TNM）分期进展和临床预后较差相关。敲低LUCAT1可抑制LUAD细胞转移和糖酵解以及肿瘤转移，而LUCAT1过表达则促进LUAD转移和糖酵解。此外，机制研究表明，LUCAT1通过海绵吸附miR-4316提高LUAD细胞转移和糖酵解，进而导致VEGFA上调。最后，在LUAD中验证了LUCAT1-miR-4316-VEGFA调控轴。我们目前的研究表明，LUCAT1作为一种竞争性内源性RNA调节miR-4316/VEGFA轴，促进LUAD细胞转移和糖酵解，为LUAD患者提供了一种新的诊断标志物和治疗靶点。
