Department of Surgical Oncology and General Surgery, The First Affiliated Hospital of China Medical University, Shenyang, China.
Cell Biochem Funct. 2019 Jan;37(1):11-20. doi: 10.1002/cbf.3368. Epub 2018 Nov 30.
Long non-coding RNAs (lncRNAs) have been demonstrated to be involved in the progression of various cancers. In this study, we aim to investigate the role of lncRNA RUNX1-IT1 in the development of colorectal cancer (CRC). The expression levels of lncRNA RUNX1-IT1 were measured using quantitative real-time Polymerase Chain Reaction(qRT-PCR). CCK8 proliferation assay, transwell assay, and flow cytometry were performed to evaluate the effect of lncRNA RUNX1-IT1 on CRC cell proliferation, migration, and apoptosis. The proliferation markers (PCNA, Ki67), apoptosis markers (cleaved-PARP, cleaved-caspase3), and MMP9 are detected by western blotting. Significant down regulation of lncRNA RUNX1-IT1 was measured in CRC tissues and three CRC cell lines (HCT116, HT29, and RKO) compared with paired nontumorous adjacent tissues (P < 0.01) or the normal colonic epithelial cell line FHC (P < 0.05), respectively. Moreover, the proliferative and migration potential of CRC cells were inhibited by overexpressing lncRNA RUNX1-IT1, which could be obviously improved by knocking down lncRNA RUNX1-IT1. The protein levels of PCNA, Ki67, and MMP9 were upregulated by overexpressing lncRNA RUNX1-IT1 and down regulated in si-RUNX1-IT1 cells. Besides, lncRNA RUNX1-IT1 could also promote the apoptosis of CRC cells. In conclusion, lncRNA RUNX1-IT1 is downregulated in CRC and plays a tumour-suppressive role due to the regulatory of cell proliferation, migration, and apoptosis. SIGNIFICANCE OF THE STUDY: We demonstrated that lncRNA RUNX1-IT1 was down regulated both in CRC tissues and cell lines. Besides, lncRNA RUNX1-IT1 could serve as a potential diagnostic biomarker and play a tumour-suppressive role owing to its good diagnostic efficacy and inhibition of CRC cell proliferation and migration.
长链非编码 RNA(lncRNA)已被证明参与了多种癌症的进展。在这项研究中,我们旨在研究 lncRNA RUNX1-IT1 在结直肠癌(CRC)发展中的作用。使用实时定量聚合酶链反应(qRT-PCR)测量 lncRNA RUNX1-IT1 的表达水平。CCK8 增殖实验、transwell 实验和流式细胞术用于评估 lncRNA RUNX1-IT1 对 CRC 细胞增殖、迁移和凋亡的影响。通过 Western blot 检测增殖标志物(PCNA、Ki67)、凋亡标志物(cleaved-PARP、cleaved-caspase3)和 MMP9。与配对的非肿瘤相邻组织(P<0.01)或正常结肠上皮细胞系 FHC(P<0.05)相比,CRC 组织和三种 CRC 细胞系(HCT116、HT29 和 RKO)中 lncRNA RUNX1-IT1 的表达水平显著下调(P<0.01)。此外,过表达 lncRNA RUNX1-IT1 抑制 CRC 细胞的增殖和迁移能力,而敲低 lncRNA RUNX1-IT1 则可明显改善这种情况。过表达 lncRNA RUNX1-IT1 可上调 PCNA、Ki67 和 MMP9 的蛋白水平,而在 si-RUNX1-IT1 细胞中则下调。此外,lncRNA RUNX1-IT1 还可以促进 CRC 细胞的凋亡。总之,lncRNA RUNX1-IT1 在 CRC 中下调,并通过调节细胞增殖、迁移和凋亡发挥肿瘤抑制作用。研究意义:我们证明 lncRNA RUNX1-IT1 在 CRC 组织和细胞系中均下调。此外,lncRNA RUNX1-IT1 可作为一种潜在的诊断生物标志物,因其具有良好的诊断功效并抑制 CRC 细胞增殖和迁移而发挥肿瘤抑制作用。