Department of Gastrointestinal Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
Department of Vascular Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, China.
Biochem Biophys Res Commun. 2021 Jun 11;557:228-235. doi: 10.1016/j.bbrc.2021.04.018. Epub 2021 Apr 19.
LncRNA-cCSC1 is highly expressed in colorectal cancer (CRC). The study was designed to evaluate the function and mechanism of lncRNA-cCSC1 in cell proliferation of CRC. RT-PCR was used to measure the expression levels of lncRNA-cCSC1 in CRC cell lines. CCK-8, colony formation, EdU staining, flow cytometry and Western blot were performed to examine the effect of interference with lncRNA-cCSC1 expression on cell proliferation. miR-124-3p and the target genes of miR-124-3p were investigated using bioinformatics analysis and verified by dual-luciferase reporter, RT-PCR and Western blot. Rescue experiments were carried out to confirm the role of miR-124-3p in cell proliferation of CRC. Our results showed that cell proliferation of CRC was promoted by lncRNA-cCSC1 upregulation and inhibited by lncRNA-cCSC1 downregulation. In addition, miR-124-3p is predicted to be the target of lncRNA-cCSC1 and is negatively correlated with lncRNA-cCSC1. Moreover, the addition of miR-124-3p mimics or inhibitor reversed the effects induced by lncRNA-cCSC1 overexpression or silencing on cell proliferation of CRC. Additionally, lncRNA-cCSC1 regulated the expression level of CD44, a target gene of miR-124-3p. Finally, we studied the effects of the lncRNA-cCSC1/miR-124-3p axis on CD44. These results indicate that lncRNA-cCSC1 promotes cell proliferation of CRC through sponging miR-124-3p and upregulating CD44.
LncRNA-cCSC1 在结直肠癌(CRC)中高表达。本研究旨在评估 lncRNA-cCSC1 在 CRC 细胞增殖中的功能和机制。RT-PCR 用于测量 CRC 细胞系中 lncRNA-cCSC1 的表达水平。CCK-8、集落形成、EdU 染色、流式细胞术和 Western blot 用于检测干扰 lncRNA-cCSC1 表达对细胞增殖的影响。生物信息学分析用于研究 miR-124-3p 及其靶基因,并用双荧光素酶报告、RT-PCR 和 Western blot 进行验证。进行挽救实验以确认 miR-124-3p 在 CRC 细胞增殖中的作用。我们的结果表明,lncRNA-cCSC1 的上调促进了 CRC 的细胞增殖,而下调则抑制了 CRC 的细胞增殖。此外,miR-124-3p 被预测为 lncRNA-cCSC1 的靶基因,并且与 lncRNA-cCSC1 呈负相关。此外,miR-124-3p 模拟物或抑制剂的添加逆转了 lncRNA-cCSC1 过表达或沉默对 CRC 细胞增殖的影响。此外,lncRNA-cCSC1 调节了 miR-124-3p 的靶基因 CD44 的表达水平。最后,我们研究了 lncRNA-cCSC1/miR-124-3p 轴对 CD44 的影响。这些结果表明,lncRNA-cCSC1 通过海绵 miR-124-3p 并上调 CD44 促进 CRC 细胞增殖。