Department of Clinical Laboratory, Yangpu Hospital, Tongji University School of Medicine, Shanghai, 200090, People's Republic of China.
Department of Clinical Laboratory, Shanghai 10th People's Hospital, Tongji University School of Medicine, No. 301 Middle Yanchang Road, Shanghai, 200072, People's Republic of China.
Hum Cell. 2020 Oct;33(4):1142-1154. doi: 10.1007/s13577-020-00393-z. Epub 2020 Aug 10.
Long noncoding RNAs (lncRNAs) are abnormally expressed in many malignant tumors and involved in regulating the malignant phenotypes of cancer cells. However, the role of LINC00665 in colorectal cancer (CRC) and its regulatory mechanism remain unclear. In this study, real-time polymerase chain reaction (RT-PCR) was used to detect the expressions of LINC00665, miR-9-5p and activating transcription factor 1 (ATF1) mRNA in CRC tissues. The expression of ATF1 in CRC tissues was also detected by immunohistochemistry and Western blot. CCK-8 and colony formation assays were employed to detect cell proliferation. Cell cycle and apoptosis were detected by flow cytometry analysis. Scratch healing assay and Transwell test were exploited to detect cell migration and invasion. The targeting relationships between LINC00665 and miR-9-5p, and miR-9-5p and ATF1 were validated by dual luciferase reporter assay. We found that LINC00665 was significantly overexpressed in CRC tissues, and it was also negatively correlated with the expression of miR-9-5p and positively associated with the expression of ATF1. Besides, LINC00665 promoted the proliferation, migration and invasion of CRC cells, and inhibited cell apoptosis by sponging miR-9-5p. ATF1 was proved to be the downstream target of miR-9-5p and was indirectly regulated by LINC00665. Collectively, it is concluded that LINC00665 contributes to the progression of CRC by regulating miR-9-5p/ATF1 axis.
长链非编码 RNA(lncRNA)在许多恶性肿瘤中异常表达,并参与调节癌细胞的恶性表型。然而,LINC00665 在结直肠癌(CRC)中的作用及其调控机制尚不清楚。在本研究中,实时聚合酶链反应(RT-PCR)用于检测 CRC 组织中 LINC00665、miR-9-5p 和激活转录因子 1(ATF1)mRNA 的表达。免疫组织化学和 Western blot 检测 CRC 组织中 ATF1 的表达。CCK-8 和集落形成实验检测细胞增殖。通过流式细胞术分析检测细胞周期和凋亡。划痕愈合实验和 Transwell 实验检测细胞迁移和侵袭。双荧光素酶报告实验验证 LINC00665 与 miR-9-5p 以及 miR-9-5p 与 ATF1 之间的靶向关系。我们发现 LINC00665 在 CRC 组织中显著过表达,并且与 miR-9-5p 的表达呈负相关,与 ATF1 的表达呈正相关。此外,LINC00665 通过海绵吸附 miR-9-5p 促进 CRC 细胞的增殖、迁移和侵袭,并抑制细胞凋亡。ATF1 被证明是 miR-9-5p 的下游靶基因,并间接受 LINC00665 调控。总之,LINC00665 通过调节 miR-9-5p/ATF1 轴促进 CRC 的进展。