Long non-coding RNA TP73-AS1 sponges miR-194 to promote colorectal cancer cell proliferation, migration and invasion via up-regulating TGFα
Long non-coding RNA TP73-AS1 sponges miR-194 to promote colorectal cancer cell proliferation, migration and invasion via up-regulating TGFα
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DOI：
10.3233/cbm-181503
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发表时间：
2018-01-01
影响因子：
3.1
通讯作者：
Cheng, Xiaohu
中科院分区：
文献类型：
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作者：
Cai, Yu;Yan, Pu;Cheng, Xiaohu
目的：结直肠癌（CRC）是全球第三大常见癌症。最近，人们发现长非编码 RNA (lncRNA) 是 CRC 进展的关键调节剂。本研究的目的是探讨lncRNA P73反义RNA 1T（TP73-AS1）在CRC发生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；进行蛋白质印迹以确定蛋白质表达水平；采用CCK-8、集落形成、伤口愈合和Transwell侵袭实验测定CRC细胞增殖、迁移和侵袭；在异种移植小鼠模型中评估体内肿瘤生长。结果：TP73-AS1 在 CRC 组织和 CRC 细胞系中均上调。 TP73-AS1 的过度表达与 CRC 患者的转移和晚期临床分期相关。 TP73-AS1过表达促进CRC细胞体外生长、增殖、迁移和侵袭；敲除TP73-AS1可显着抑制体外CRC细胞的生长、增殖、迁移和侵袭以及体内肿瘤的生长。生物信息学分析和荧光素酶报告基因检测表明TP73-AS1可以直接与miR-194结合，并且TP73-AS1负向调节CRC细胞中miR-194的表达。进一步研究表明，miR-194通过靶向其3'非翻译区负向调节转化生长因子α（TGF α）的下游靶标，而TP73-AS1正向调节结直肠癌细胞中TGF α的表达。此外，miR-194的过表达抑制了CRC细胞的增殖和侵袭，并减弱了TP73-AS1过表达对CRC细胞增殖和侵袭的影响。 TGF α的沉默抑制了CRC细胞的增殖和侵袭，并且还逆转了TP73-AS1过表达对CRC细胞增殖和侵袭的影响。结论：本研究证明TP73-AS1通过作为竞争性内源RNA海绵miR-194来调节TGF α的表达，从而调节CRC进展。
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 (TGF alpha) via targeting its 3' untranslated region, and TP73-AS1 positively regulated the expression of TGF alpha 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 TGF alpha 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 TGF alpha.