LINC01605 通过靶向 miR-3960/SOX11 调控结直肠癌细胞的增殖、迁移和侵袭。

LINC01605 regulates proliferation, migration and invasion of colorectal cancer cells via targeting miR-3960/SOX11.

机构信息

Department of Gastroenterology, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Zhengzhou, China.

出版信息

Eur Rev Med Pharmacol Sci. 2021 Feb;25(3):1322-1329. doi: 10.26355/eurrev_202102_24837.

DOI:10.26355/eurrev_202102_24837

PMID:33629302

Abstract

OBJECTIVE

We aimed to determine the expression level of long intergenic non-coding ribonucleic acid 1605 (LINC01605) in colorectal cancer (CRC), and to explore the effects of the LINC01605/microRNA (miR)-3960/sex-determining region Y-box 11 (SOX11) regulatory axis on the biological behaviors of CRC cells and the molecular mechanism therein.

PATIENTS AND METHODS

Tissue specimens were collected from 38 patients with CRC, and the relative expression level of LINC01605 in the CRC tissues and CRC cells was measured using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Then, the effects of LINC01605 on the proliferation, apoptosis, invasion and metastasis of CRC cells were observed via in vitro assays [cell counting kit (CCK)-8 assay, flow cytometry and transwell assay]. Besides, the possible miRNAs binding to LINC01605 were predicted by the bioinformatics method, and they were screened and verified using qRT-PCR and Dual-Luciferase reporter gene assay. Finally, the downstream target genes of miR-3960 were predicted by means of bioinformatics, and they were also screened and confirmed via qRT-PCR and Dual-Luciferase reporter gene assay.

RESULTS

According to the results of qRT-PCR, the expression of LINC01605 was up-regulated in 31 out of 38 cases of CRC tissue specimens, and its expression in CRC cells was higher than that in normal colorectal cells. The results of in vitro assays revealed that the proliferation, migration and invasion abilities of CRC cells were weakened, with an increased apoptosis rate after interference with LINC01605 expression. Based on the results of qRT-PCR and Dual-Luciferase reporter gene assay, miR-3960 was the target of LINC01605, while SOX11 was the target of miR-3960. Moreover, the expression of miR-3960 rose, but that of SOX11 declined after interference with LINC01605 expression. It was found through Western blotting that the protein expression of SOX11 was lowered after interference with LINC01605 expression.

CONCLUSIONS

LINC01605 has an up-regulated expression in CRC, and accelerates the proliferation, migration and metastasis of CRC cells by the miR-3960/SOX11 regulatory axis.

摘要

目的

本研究旨在确定长链非编码 RNA 1605（LINC01605）在结直肠癌（CRC）中的表达水平，并探讨 LINC01605/微小 RNA（miR）-3960/性别决定区 Y 框 11（SOX11）调控轴对 CRC 细胞生物学行为的影响及其内在分子机制。

患者与方法

收集 38 例 CRC 患者的组织标本，采用实时定量逆转录聚合酶链反应（qRT-PCR）检测 CRC 组织和 CRC 细胞中 LINC01605 的相对表达水平。然后，通过细胞计数试剂盒（CCK-8）检测、流式细胞术和 Transwell 检测观察 LINC01605 对 CRC 细胞增殖、凋亡、侵袭和转移的影响。此外，通过生物信息学方法预测可能与 LINC01605 结合的 miRNA，并通过 qRT-PCR 和双荧光素酶报告基因检测进行筛选和验证。最后，通过生物信息学方法预测 miR-3960 的下游靶基因，并通过 qRT-PCR 和双荧光素酶报告基因检测进行筛选和确认。

结果

qRT-PCR 结果显示，38 例 CRC 组织标本中 31 例 LINC01605 表达上调，CRC 细胞中 LINC01605 的表达高于正常结肠直肠细胞。体外实验结果表明，干扰 LINC01605 表达后，CRC 细胞的增殖、迁移和侵袭能力减弱，凋亡率增加。基于 qRT-PCR 和双荧光素酶报告基因检测的结果，miR-3960 是 LINC01605 的靶基因，而 SOX11 是 miR-3960 的靶基因。此外，干扰 LINC01605 表达后，miR-3960 的表达上调，而 SOX11 的表达下调。Western blot 结果显示，干扰 LINC01605 表达后 SOX11 蛋白表达降低。