Suzhou Hospital of Traditional Chinese Medicine, Suzhou, 215009, Jiangsu, China.
Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou, 450008, Henan, China.
Eur J Pharmacol. 2019 Sep 5;858:172464. doi: 10.1016/j.ejphar.2019.172464. Epub 2019 Jun 22.
Long intergenic non-coding RNA No.668 (LINC00668) is implicated in the development of various malignancies. However, the role of LINC00668 and underlying mechanism in colorectal cancer (CRC) remains totally unknown. The expression pattern of LINC00668 in CRC cells were determined by qRT-PCR. CCK-8, EdU incorporation, flow cytometry, Transwell, and wound-healing assays were run to evaluate the functions of LINC00668 in CRC cells. Bioinformatics analyses were used to identify the LINC00668-specific binding with miRNAs that were screened by RNA pull-down. RNA immunoprecipitation and luciferase gene report assay were performed to confirm the interaction between miR-188-5p and LINC00668 in CRC cells. LINC00668 was significantly upregulated in CRC tissues and cells. Knockdown of LINC00668 suppressed cell proliferation and migration potential and induced cell apoptosis, but inhibition of miR-188-5p which was predicted to bind with LINC00668 reversed these effects. Furthermore, USP47 was a direct target of miR-188-5p, and overexpression of USP47 attenuated LINC00668 knockdown-induced tumor suppressive effects in CRC cells. Conclusively, our findings demonstrated that lncRNA LINC00668 acted as an oncogenic role in CRC cells by sponging miR-188-5p and upregulating USP47 and may represent a potential marker for CRC patients.
长链非编码 RNA No.668（LINC00668）参与多种恶性肿瘤的发生发展。然而，LINC00668 在结直肠癌（CRC）中的作用及其潜在机制尚不清楚。通过 qRT-PCR 检测 CRC 细胞中 LINC00668 的表达模式。通过 CCK-8、EdU 掺入、流式细胞术、Transwell 和划痕愈合实验评估 LINC00668 在 CRC 细胞中的功能。通过 RNA 下拉筛选出 miRNA 的生物信息学分析，确定 LINC00668 与 miRNA 的特异性结合。RNA 免疫沉淀和荧光素酶基因报告实验证实 miR-188-5p 与 LINC00668 在 CRC 细胞中的相互作用。LINC00668 在 CRC 组织和细胞中显著上调。敲低 LINC00668 抑制细胞增殖和迁移能力，并诱导细胞凋亡，但抑制 miR-188-5p 可逆转这些作用，miR-188-5p 被预测与 LINC00668 结合。此外，USP47 是 miR-188-5p 的直接靶标，过表达 USP47 减弱了 LINC00668 敲低对 CRC 细胞中肿瘤抑制作用。总之，我们的研究结果表明，lncRNA LINC00668 通过海绵 miR-188-5p 上调 USP47 在 CRC 细胞中发挥致癌作用，可能成为 CRC 患者的潜在标志物。
