Zhang Yueyan, Liu Xingchi, Li Qiang, Zhang Yong
Department of Pathology, Liaoning Cancer Hospital and Institute, Cancer Hospital of China Medical University, Shenyang, People's Republic of China,
Department of Thoracic Surgery, General Hospital of Shenyang Military Command, Shenyang, People's Republic of China.
Cancer Manag Res. 2019 Feb 22;11:1779-1789. doi: 10.2147/CMAR.S192452. eCollection 2019.
lncRNAs are widely involved in multiple malignancies including colorectal cancer (CRC). The expression and function of long intergenic non-protein coding RNA 460 (LINC00460) in CRC remains obscure.
In the present study, quantitative real-time PCR assays were applied to detect the expression changes of LINC00460 and microRNA-939-5p (miR-939-5p) in CRC tissue specimens and cell lines. Western blot assays were used to measure the changes of LIMK2. Bioinformatics analysis, luciferase assays, and RNA pull-down assays were applied to determine the targeting binding effect between LINC00460 and miR-939-5p as well as LIMK2 and miR-939-5p. Transwell assays were used to evaluate the metastatic ability changes of CRC line HT29 and LOVO cells.
We found that LINC00460 was upregulated and closely correlated to clinicopathological features and poor prognosis of patients with CRC. Functionally, we elucidated that LINC00460 promoted metastasis in CRC cell lines HT29 and LOVO. Further, we showed that LIMK2 was a downstream effector in the LINC00460-induced promotion of metastasis in CRC cells HT29 and LOVO. Through online bioinformatics analysis, LINC00460 and LIMK2 were demonstrated to share similar microRNA response elements for miR-939-5p. Then, LINC00460 and LIMK2 were verified to be the targets of miR-939-5p via a luciferase assay and an RNA pull-down assay. Also, miR-939-5p was showed to suppress metastasis by targeting of LIMK2. Lastly, we revealed that LINC00460 promoted LIMK2-mediated metastasis via miR-939-5p sponging in CRC cells HT29 and LOVO.
The findings of this study showed that LINC00460 works as an oncogene in CRC and promoted CRC cell metastasis via regulation of miR-939-3p/LIMK2 axial. The present study might provide a new target in treating CRC.
长链非编码RNA（lncRNAs）广泛参与包括结直肠癌（CRC）在内的多种恶性肿瘤。长链基因间非编码RNA 460（LINC00460）在结直肠癌中的表达及功能仍不清楚。
在本研究中，应用定量实时PCR检测LINC00460和微小RNA-939-5p（miR-939-5p）在结直肠癌组织标本和细胞系中的表达变化。采用蛋白质免疫印迹法检测LIMK2的变化。应用生物信息学分析、荧光素酶报告基因检测和RNA下拉实验确定LINC00460与miR-939-5p以及LIMK2与miR-939-5p之间的靶向结合作用。采用Transwell实验评估结直肠癌细胞系HT29和LOVO细胞转移能力的变化。
我们发现LINC00460上调，且与结直肠癌患者的临床病理特征及不良预后密切相关。在功能上，我们阐明LINC00460促进结直肠癌细胞系HT29和LOVO的转移。此外，我们发现LIMK2是LINC00460诱导结直肠癌细胞系HT29和LOVO转移的下游效应分子。通过在线生物信息学分析，发现LINC00460和LIMK2共享相似的miR-939-5p微小RNA反应元件。随后，通过荧光素酶报告基因检测和RNA下拉实验证实LINC00460和LIMK2是miR-939-5p的靶点。同时，发现miR-939-5p通过靶向LIMK2抑制转移。最后，我们揭示LINC00460通过在结直肠癌细胞系HT29和LOVO中充当miR-939-5p的海绵，促进LIMK2介导的转移。
本研究结果表明，LINC00460在结直肠癌中作为癌基因发挥作用，并通过调节miR-939-3p/LIMK2轴促进结直肠癌细胞转移。本研究可能为结直肠癌的治疗提供新靶点。
