通过靶向 miR-548b-3p 下调 CPSF6,抑制 circ_0000673 可阻止结直肠癌的进展。

IF 2.1 4区 医学 Q3 MEDICINE, RESEARCH & EXPERIMENTAL
Shuang Li, Tuoyun Yang, Lu Liu, Baorong Hu, Xi Chen, Wenting Zhao, Xin Hai
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引用次数: 0

摘要

背景:大肠癌(CRC)是最常见的癌症之一,其进展受多种因素调控,包括环状 RNA(circRNA):本研究旨在确定 circ_0000673 在 CRC 中的作用:我们使用实时定量聚合酶链反应(qPCR)检测了DLD-1和RKO细胞中circ_0000673、miR-548b-3p以及裂解和多腺苷酸化特异性因子6(CPSF6)的表达。细胞计数试剂盒-8(CCK-8)和 5-乙炔基-2'-脱氧尿苷(EdU)测定法用于确定 circ_0000673 在增殖中的作用。伤口愈合和透孔试验用于检测细胞迁移和侵袭能力。用 Western 印迹法检测了 CPSF6 蛋白和干细胞相关蛋白的表达。通过双荧光素酶报告实验验证了 miR-548b-3p 与 circ_0000673 或 CPSF6 之间的推测关系。还通过裸鼠肿瘤异种移植实验研究了 circ_0000673 在 CRC 中的作用:结果:Circ_0000673 在 CRC 组织和癌细胞中的表达增加。沉默 circ_0000673 可减少肿瘤细胞的增殖、迁移和侵袭,同时降低细胞的干性。研究发现,miR-548b-3p 是 circ_0000673 的靶点,而 CPSF6 是 miR-548b-3p 的下游靶点。在拯救试验中,si-circ_0000673、抗-miR-548b-3p 和 oe-CPSF6 的肿瘤调节作用分别被抗-miR-548b-3p、si-CPSF6 和 si-circ_0000673 部分逆转。下调circ_0000673可减少实体瘤在体内的生长:Circ_0000673抑制剂通过靶向miR-548b-3p减少了CPSF6的表达,从而阻断了CRC肿瘤细胞的增殖、迁移和侵袭。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Inhibiting circ_0000673 blocks the progression of colorectal cancer through downregulating CPSF6 via targeting miR-548b-3p.

Background: Colorectal cancer (CRC) is one of the most common cancers, and its progression is regulated by several factors, including circular RNA (circRNA).

Objectives: The objective of this study was to determine the role, or roles, of circ_0000673 in CRC.

Material and methods: We used quantitative real-time polymerase chain reaction (qPCR) to detect the expression of circ_0000673, miR-548b-3p and cleavage and polyadenylation specific factor 6 (CPSF6) in DLD-1 and RKO cells. Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) assays were used to determine circ_0000673 roles in proliferation. Wound healing and transwell assays were used to detect cell migration and invasion abilities. Expression of CPSF6 protein and stem cell-associated proteins were examined using western blot. The putative relationship between miR-548b-3p and circ_0000673 or CPSF6 was verified with dual-luciferase reporter assay. The role of circ_0000673 in CRC was also investigated in a tumor xenograft assay in nude mice.

Results: Circ_0000673 expression was increased in CRC tissues and cancer cells. Silencing circ_0000673 reduced tumor cell proliferation, migration and invasion, while also decreasing cell stemness. MiR-548b-3p was found to be a target of circ_0000673, while CPSF6 was a downstream target of miR-548b-3p. The tumor-regulatory effects of si-circ_0000673, anti-miR-548b-3p and oe-CPSF6 were partially reversed by anti-miR-548b-3p, si-CPSF6 and si-circ_0000673, respectively, in rescue assays. Downregulation of circ_0000673 reduced solid tumor growth in vivo.

Conclusions: Circ_0000673 inhibition reduced CPSF6 expression by targeting miR-548b-3p, thereby blocking proliferation, migration and invasion of CRC tumor cells.

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来源期刊
Advances in Clinical and Experimental Medicine
Advances in Clinical and Experimental Medicine MEDICINE, RESEARCH & EXPERIMENTAL-
CiteScore
3.70
自引率
4.80%
发文量
153
审稿时长
6-12 weeks
期刊介绍: Advances in Clinical and Experimental Medicine has been published by the Wroclaw Medical University since 1992. Establishing the medical journal was the idea of Prof. Bogumił Halawa, Chair of the Department of Cardiology, and was fully supported by the Rector of Wroclaw Medical University, Prof. Zbigniew Knapik. Prof. Halawa was also the first editor-in-chief, between 1992-1997. The journal, then entitled "Postępy Medycyny Klinicznej i Doświadczalnej", appeared quarterly. Prof. Leszek Paradowski was editor-in-chief from 1997-1999. In 1998 he initiated alterations in the profile and cover design of the journal which were accepted by the Editorial Board. The title was changed to Advances in Clinical and Experimental Medicine. Articles in English were welcomed. A number of outstanding representatives of medical science from Poland and abroad were invited to participate in the newly established International Editorial Staff. Prof. Antonina Harłozińska-Szmyrka was editor-in-chief in years 2000-2005, in years 2006-2007 once again prof. Leszek Paradowski and prof. Maria Podolak-Dawidziak was editor-in-chief in years 2008-2016. Since 2017 the editor-in chief is prof. Maciej Bagłaj. Since July 2005, original papers have been published only in English. Case reports are no longer accepted. The manuscripts are reviewed by two independent reviewers and a statistical reviewer, and English texts are proofread by a native speaker. The journal has been indexed in several databases: Scopus, Ulrich’sTM International Periodicals Directory, Index Copernicus and since 2007 in Thomson Reuters databases: Science Citation Index Expanded i Journal Citation Reports/Science Edition. In 2010 the journal obtained Impact Factor which is now 1.179 pts. Articles published in the journal are worth 15 points among Polish journals according to the Polish Committee for Scientific Research and 169.43 points according to the Index Copernicus. Since November 7, 2012, Advances in Clinical and Experimental Medicine has been indexed and included in National Library of Medicine’s MEDLINE database. English abstracts printed in the journal are included and searchable using PubMed http://www.ncbi.nlm.nih.gov/pubmed.
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