Table_10_lnc-REG3G-3-1/miR-215-3p Promotes Brain Metastasis of Lung Adenocarcinoma by Regulating Leptin and SLC2A5.DOC

This study aims to explore the role and mechanism of specific lncRNA in brain metastasis (BM) from lung adenocarcinoma (LADC), providing an effective biomarker for early diagnosis and targeted therapy of BM from LADC. Based on the gene expression profiles of lncRNA and mRNA in LADC and BM tissues detected by Gene Chip, lnc-REG3G-3-1 was selected, and the related genes, including miR-215-3p, leptin, and SLC2A5, were identified by data analysis. Human LADC cell lines A549 and H1299 were cultured. Dual-luciferase and endogenous validation experiments were used to confirm the regulation between these genes. Real-time quantitative reverse transcription–polymerase chain reaction and Western blotting were used to detect gene expression. The tumor metastasis-related gene function of lnc-REG3G-3-1 and miR-215-3p in H1299 cells was verified by Transwell invasion, migration assays, and scratch testing. Nude mice xenograft tumors constructed with decreased lnc-REG3G-3-1 confirmed the influences on gene expression in vivo. lnc-REG3G-3-1 was highly expressed in BM tissues that originated from LADC compared with that in primary cancer tissues. lnc-REG3G-3-1 reduced miR-215-3p expression, thereby regulating the target genes leptin and SLC2A5 and the signaling pathways, taking part in the lnc-REG3G-3-1/miR-215-3p axis in the process of BM from LADC. lnc-REG3G-3-1, leptin, and SLC2A5 through regulating signaling pathways may be jointly involved in the regulation of the biological process of BM in patients with LADC.

本研究旨在探讨特定长非编码RNA (lncRNA) 在肺腺癌 (LADC) 脑转移 (BM) 中的作用及其机制，以期为LADC BM的早期诊断和靶向治疗提供有效的生物标志物。基于对LADC及BM组织中lncRNA和mRNA基因表达谱的基因芯片检测，通过数据分析选出了lnc-REG3G-3-1，并鉴定了包括miR-215-3p、瘦素和SLC2A5在内的相关基因。培养人LADC细胞系A549和H1299。利用双荧光素酶和内源验证实验确认了这些基因之间的调控关系。通过实时定量逆转录聚合酶链反应和蛋白质印迹检测基因表达。通过Transwell侵袭、迁移实验和划痕实验验证了lnc-REG3G-3-1和miR-215-3p在H1299细胞中的肿瘤转移相关基因功能。构建的降低lnc-REG3G-3-1表达的裸鼠异种移植肿瘤模型证实了其在体内对基因表达的影响。与原发肿瘤组织相比，源于LADC的BM组织中lnc-REG3G-3-1的表达显著升高。lnc-REG3G-3-1通过降低miR-215-3p的表达，进而调节靶基因瘦素和SLC2A5及信号通路，参与构成lnc-REG3G-3-1/miR-215-3p轴，在LADC BM的过程中发挥作用。lnc-REG3G-3-1、瘦素和SLC2A5通过调节信号通路，可能共同参与LADC患者肿瘤转移生物学过程的调控。