Liu Chang, Hou Jianfeng, Shan Fengxiao, Wang Lijuan, Lu Hanjie, Ren Tiejun
Department of Oncology, Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang 471000, People's Republic of China.
Onco Targets Ther. 2020 Jun 2;13:4931-4942. doi: 10.2147/OTT.S237580. eCollection 2020.
Long non-coding RNA colorectal neoplasia differentially expressed (lncRNA CRNDE) and microRNA-126-5p (miR-126-5p) were reported to be related to the development of colorectal carcinoma (CRC). However, the detailed mechanism of CRNDE and miR-126-5p is not fully understood. The purpose of this research was to explore their roles and molecular mechanism in CRC.
Quantitative real-time polymerase chain reaction was performed to detect the transcription levels of genes. Paclitaxel (PTX) was used to analyze cell drug resistance. 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay and flow cytometry analysis were employed to assess cell proliferation and apoptosis, respectively. Furthermore, cell migratory and invasive abilities were measured using transwell assay. The interaction between miR-126-5p and CRNDE or ATPase family AAA domain-containing protein 2 (ATAD2) was predicted by online tool starbase and then confirmed using the dual-luciferase reporter assay. Besides, Western blot assay was carried out to detect the levels of proteins.
CRNDE and ATAD2 expressions were upregulated and miR-126-5p expression was downregulated in CRC tissues and cells. CRNDE depletion repressed PTX resistance and the growth of CRC cells. Interestingly, we found that miR-126-5p was a target gene of CRNDE, and miR-126-5p directly targeted ATAD2. Furthermore, CRNDE affected CRC cell progression via modulation of miR-126-5p/ATAD2 axis in CRC cells.
Our data suggested that CRNDE regulated CRC cell development and PTX resistance by modulating miR-126-5p/ATAD2 axis, providing the theoretical basis for the treatment of CRC patients.
长链非编码RNA结直肠肿瘤差异表达基因（lncRNA CRNDE）和微小RNA-126-5p（miR-126-5p）据报道与结直肠癌（CRC）的发生发展有关。然而，CRNDE和miR-126-5p的详细机制尚未完全明确。本研究旨在探讨它们在CRC中的作用及分子机制。
采用定量实时聚合酶链反应检测基因转录水平。使用紫杉醇（PTX）分析细胞耐药性。分别采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法和流式细胞术分析评估细胞增殖和凋亡。此外，使用Transwell实验检测细胞迁移和侵袭能力。通过在线工具starbase预测miR-126-5p与CRNDE或含ATP酶家族AAA结构域蛋白2（ATAD2）之间的相互作用，然后用双荧光素酶报告基因实验进行验证。此外，进行蛋白质免疫印迹实验检测蛋白质水平。
CRC组织和细胞中CRNDE和ATAD2表达上调，miR-126-5p表达下调。CRNDE缺失可抑制PTX耐药性及CRC细胞生长。有趣的是，我们发现miR-126-5p是CRNDE的靶基因，且miR-126-5p直接靶向ATAD2。此外，CRNDE通过调节CRC细胞中的miR-126-5p/ATAD2轴影响CRC细胞进展。
我们的数据表明，CRNDE通过调节miR-126-5p/ATAD2轴调控CRC细胞发育和PTX耐药性，为CRC患者的治疗提供了理论依据。
