长链非编码 RNA TP73-AS1 通过海绵吸附 miR-194 促进结直肠癌细胞增殖、迁移和侵袭并上调 TGFα。

Long non-coding RNA TP73-AS1 sponges miR-194 to promote colorectal cancer cell proliferation, migration and invasion via up-regulating TGFα.

机构信息

Department of General Surgery, The First Affiliated Hospital of Xi'an Medical University, Xi'an, Shaanxi, China.

Department of General Surgery, Shaanxi Sengong Hospital, Xi'an, Shaanxi, China.

出版信息

Cancer Biomark. 2018;23(1):145-156. doi: 10.3233/CBM-181503.

DOI:10.3233/CBM-181503

PMID:30010111

Abstract

OBJECTIVE

Colorectal cancer (CRC) is the 3rd most common cancer worldwide. Recently, long non-coding RNAs (lncRNAs) were found to be critical modulators in the CRC progression. The aim of this study is to investigate the potential roles of lncRNA P73 antisense RNA 1T (TP73-AS1) in CRC development and progression.

METHODS

Quantitative real-time PCR (qRT-PCR) was performed to determine relevant gene expression levels; western blot was performed to determine protein expression levels; CCK-8, colony formation, wound healing and Transwell invasion assays were used to determined CRC cell proliferation, migration and invasion; in vivo tumor growth was assessed in xenograft mice model.

RESULTS

TP73-AS1 was up-regulated in both CRC tissues and CRC cell lines. Overexpression of TP73-AS1 was associated with metastasis and advanced clinical stages in CRC patients. Overexpression of TP73-AS1 promoted CRC cell growth, proliferation, migration and invasion in vitro; and knockdown of TP73-AS1 significantly inhibited CRC cell growth, proliferation, migration and invasion in vitro as well as tumor growth in vivo. Bioinformatics analysis and luciferase reporter assay indicated that TP73-AS1 could bind directly with miR-194, and TP73-AS1 negatively regulated the expression of miR-194 in CRC cells. Further study indicated that miR-194 negatively regulated the downstream target of transforming growth factor alpha (TGFα) via targeting its 3' untranslated region, and TP73-AS1 positively regulated the expression of TGFα in CRC cells. Moreover, overexpression of miR-194 suppressed CRC cell proliferation and invasion, and attenuated the effects of TP73-AS1 overexpression on CRC cell proliferation and invasion. Silence of TGFα inhibited CRC cell proliferation and invasion, and also reversed the effects of TP73-AS1 overexpression on CRC cell proliferation and invasion.

CONCLUSIONS

this study demonstrated that TP73-AS1 regulated CRC progression by acting as a competitive endogenous RNA to sponge miR-194 to modulate the expression of TGFα.

摘要

目的

结直肠癌（CRC）是全球第三大常见癌症。最近，长非编码 RNA（lncRNA）被发现是 CRC 进展的关键调节因子。本研究旨在探讨 lncRNA P73 反义 RNA 1T（TP73-AS1）在 CRC 发展和进展中的潜在作用。

方法

采用实时定量 PCR（qRT-PCR）测定相关基因表达水平；采用 Western blot 测定蛋白表达水平；采用 CCK-8、集落形成、划痕愈合和 Transwell 侵袭实验测定 CRC 细胞增殖、迁移和侵袭能力；在异种移植小鼠模型中评估体内肿瘤生长情况。

结果

TP73-AS1 在 CRC 组织和 CRC 细胞系中均上调。TP73-AS1 的过表达与 CRC 患者的转移和晚期临床分期相关。TP73-AS1 的过表达促进 CRC 细胞在体外的生长、增殖、迁移和侵袭；而 TP73-AS1 的敲低显著抑制 CRC 细胞在体外的生长、增殖、迁移和侵袭以及体内的肿瘤生长。生物信息学分析和荧光素酶报告实验表明，TP73-AS1 可以直接与 miR-194 结合，并且 TP73-AS1 负调控 CRC 细胞中 miR-194 的表达。进一步的研究表明，miR-194 通过靶向其 3'非翻译区负调控转化生长因子 alpha（TGFα）的下游靶标，而 TP73-AS1 正调控 CRC 细胞中 TGFα 的表达。此外，过表达 miR-194 抑制 CRC 细胞的增殖和侵袭，并减弱 TP73-AS1 过表达对 CRC 细胞增殖和侵袭的影响。沉默 TGFα 抑制 CRC 细胞的增殖和侵袭，并逆转 TP73-AS1 过表达对 CRC 细胞增殖和侵袭的影响。