Yin Dianhe, Hua Li, Wang Jiao, Liu Yuru, Li Xiaoyan
Department of General Practice, Huaihe Hospital of Henan University, Kaifeng 475000, Henan, People's Republic of China.
Department of Respiratory and Critical Care Medicine, Peking University International Hospital, Beijing, People's Republic of China.
Onco Targets Ther. 2020 Jul 22;13:7111-7123. doi: 10.2147/OTT.S243542. eCollection 2020.
Long non-coding RNAs (lncRNAs) were confirmed to play important roles in human cancers. In this study, we explored the functional role of lncRNA double homeobox A pseudogene 8 (DUXAP8) in non-small-cell lung cancer (NSCLC).
Real-time quantitative PCR (RT-qPCR) was used to detect DUXAP8 and microRNA-409-3p (miR-409-3p) expression. CCK-8, cell colony formation assay, and Transwell migration assay were performed to measure cell growth and migration, respectively. The expression of the relative proteins was detected by Western blot. Cell glycolysis was determined by glucose uptake, adenosine triphosphate (ATP) concentration, lactate generation, extracellular acidification rate and oxygen consumption rate assays. Bioinformatics analysis and dual-luciferase reporter assay were used to measure the interaction among DUXAP8, miR-409-3p, hexokinase 2 (HK2) and lactate dehydrogenase A (LDHA). In vivo, subcutaneous tumor formation assay was performed in the nude mice.
DUXAP8 was highly expressed in NSCLC, while miR-409-3p was downregulated. High expression of DUXAP8 was positively related to the grade division and negatively associated with the 5-year survival rate of NSCLC patients. Downregulated DUXAP8 significantly suppressed cell growth, metastasis and glycolysis. Besides, DUXAP8 sponged miR-409-3p to promote HK2 and LDHA expression. DUXAP8 promoted cell viability, migration and glycolysis by regulating miR-409-3p/HK2/LDHA axis. Moreover, DUXAP8 downregulation markedly inhibited tumor growth in vivo.
Our findings demonstrated that DUXAP8 served as an oncogene in the progression of NSCLC.
长链非编码RNA（lncRNAs）已被证实在人类癌症中发挥重要作用。在本研究中，我们探讨了lncRNA双同源盒A假基因8（DUXAP8）在非小细胞肺癌（NSCLC）中的功能作用。
采用实时定量PCR（RT-qPCR）检测DUXAP8和微小RNA-409-3p（miR-409-3p）的表达。分别进行CCK-8、细胞集落形成试验和Transwell迁移试验以检测细胞生长和迁移情况。通过蛋白质印迹法检测相关蛋白的表达。通过葡萄糖摄取、三磷酸腺苷（ATP）浓度、乳酸生成、细胞外酸化率和耗氧率试验测定细胞糖酵解情况。采用生物信息学分析和双荧光素酶报告基因试验检测DUXAP8、miR-409-3p、己糖激酶2（HK2）和乳酸脱氢酶A（LDHA）之间的相互作用。在体内，对裸鼠进行皮下肿瘤形成试验。
DUXAP8在NSCLC中高表达，而miR-409-3p下调。DUXAP8的高表达与NSCLC患者的分级呈正相关，与5年生存率呈负相关。下调DUXAP8可显著抑制细胞生长、转移和糖酵解。此外，DUXAP8通过吸附miR-409-3p来促进HK2和LDHA的表达。DUXAP8通过调节miR-409-3p/HK2/LDHA轴促进细胞活力、迁移和糖酵解。此外，下调DUXAP8可显著抑制体内肿瘤生长。
我们的研究结果表明，DUXAP8在NSCLC进展中作为一种癌基因发挥作用。
