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长非编码 RNA RUNX1-IT1 通过抑制细胞增殖和迁移在结直肠癌中发挥肿瘤抑制作用。

Long non-coding RNA RUNX1-IT1 plays a tumour-suppressive role in colorectal cancer by inhibiting cell proliferation and migration.

机构信息

Department of Surgical Oncology and General Surgery, The First Affiliated Hospital of China Medical University, Shenyang, China.

出版信息

Cell Biochem Funct. 2019 Jan;37(1):11-20. doi: 10.1002/cbf.3368. Epub 2018 Nov 30.

Abstract

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.

摘要

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。

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