Hsa_circ_0007534 敲低通过调控 miR-613/SLC25A22 轴抑制结直肠癌细胞的发展。

Hsa_circ_0007534 knockdown represses the development of colorectal cancer cells through regulating miR-613/SLC25A22 axis.

机构信息

Department of Gastrointestinal Colorectal Surgery, China-Japan Union Hospital, Jilin University, Changchun, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Mar;24(6):3004-3022. doi: 10.26355/eurrev_202003_20665.

DOI:10.26355/eurrev_202003_20665

PMID:32271418

Abstract

OBJECTIVE

Colorectal cancer (CRC) is a common tumor around the world. Circular RNAs (circRNAs) have been reported to be related to the development of CRC. However, the detailed mechanism is complicated. This study aimed to reveal the functional mechanism of circ_0007534 in CRC.

PATIENTS AND METHODS

Quantitative real time polymerase chain reation (qRT-PCR) and Western blot assay were performed to analyze gene expression. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and colony formation assay were carried out to determine cell proliferation ability. Furthermore, cell migratory and invasive abilities were assessed by transwell assay. Glycolytic metabolism was examined via the measurements of extracellular acidification rate (ECAR), glucose consumption, and lactate production. Also, the interaction between circ_0007534 or solute carrier family 25 member 22 (SLC25A22) and miR-613 was predicted and confirmed by starBase v2.0 and the Dual-Luciferase reporter assay, respectively. Mouse xenograft was performed to investigate the effect of circ_0007534 on tumor growth in vivo.

RESULTS

Circ_0007534 and SLC25A22 levels were upregulated, and miR-613 level was downregulated in CRC tissues/cells. Circ_0007534 knockdown repressed CRC cell proliferation, colony formation, migration, invasion, and glycolysis. Interestingly, Circ_0007534 targeted miR-613, and miR-613 targeted SLC25A22. Circ_0007534 exerted its function by repressing miR-613 expression, and miR-613 exerted its function via inhibiting SLC25A22 expression. Also, Circ_0007534 repressed miR-613 expression to upregulate SLC25A22 level. Circ_0007534 depletion repressed tumor growth in vivo.

CONCLUSIONS

We demonstrated that circ_0007534 knockdown suppressed the growth of CRC cells by regulating miR-613/SLC25A22 axis, providing potential target for the treatment of CRC.

摘要

目的

结直肠癌（CRC）是一种常见的肿瘤。已经有报道表明环状 RNA（circRNAs）与 CRC 的发展有关。然而，其详细的机制很复杂。本研究旨在揭示 circ_0007534 在 CRC 中的功能机制。

患者与方法

采用实时定量聚合酶链反应（qRT-PCR）和 Western blot 检测分析基因表达。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法和集落形成实验检测细胞增殖能力。此外，通过 Transwell 实验检测细胞迁移和侵袭能力。通过测量细胞外酸化率（ECAR）、葡萄糖消耗和乳酸生成来检测糖酵解代谢。另外，通过 starBase v2.0 预测和 Dual-Luciferase 报告实验分别验证 circ_0007534 与溶质载体家族 25 成员 22（SLC25A22）和 miR-613 之间的相互作用。通过小鼠异种移植实验研究 circ_0007534 对体内肿瘤生长的影响。

结果

CRC 组织/细胞中 circ_0007534 和 SLC25A22 水平上调，miR-613 水平下调。circ_0007534 敲低抑制 CRC 细胞增殖、集落形成、迁移、侵袭和糖酵解。有趣的是，circ_0007534 靶向 miR-613，而 miR-613 靶向 SLC25A22。circ_0007534 通过抑制 miR-613 的表达发挥其功能，而 miR-613 通过抑制 SLC25A22 的表达发挥其功能。此外，circ_0007534 抑制 miR-613 的表达以上调 SLC25A22 水平。circ_0007534 耗竭抑制体内肿瘤生长。