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长链非编码RNA牛磺酸上调基因1(TUG1)下调通过miR-498调控细胞分裂周期蛋白42(CDC42)的表达来抑制食管鳞状细胞癌细胞的增殖和侵袭。

Long Non-Coding RNA Taurine Upregulated Gene 1 (TUG1) Downregulation Constrains Cell Proliferation and Invasion through Regulating Cell Division Cycle 42 (CDC42) Expression Via MiR-498 in Esophageal Squamous Cell Carcinoma Cells.

作者信息

Wang Zhifeng, Liu Jingmei, Wang Rong, Wang Qinqin, Liang Rong, Tang Jinliang

机构信息

Department of Digestive Endoscopy, Shanxi Provincial People's Hospital, Taiyuan, Shanxi, China (mainland).

Department of Gastroenterology, Shanxi Cancer Hospital, Taiyuan, Shanxi, China (mainland).

出版信息

Med Sci Monit. 2020 Mar 6;26:e919714. doi: 10.12659/MSM.919714.

Abstract

BACKGROUND Esophageal squamous cell carcinoma (ESCC) is a malignant tumor of the gastrointestinal tract. Taurine upregulated gene 1 (TUG1), a long non-coding (lnc) RNA, also known as LIN00080 or TI-227H, was connected with the tumorigenesis of various diseases. Hence, we plumed the role and molecular mechanism of TUG1 in the progression of ESCC. MATERIAL AND METHODS Expression patterns of TUG1, microRNA-498 (miR-498), and cell division cycle 42 (CDC42) mRNA were assessed using quantitative real time polymerase chain reaction (qRT-PCR). The expression level of CDC42 protein was evaluated via western blot analysis. Cell proliferation and invasion were determined with Cell Counting Kit-8 (CCK-8) assay or Transwell assay. The relationship between miR-498 and TUG1 or CDC42 was predicted by online bioinformatics database LncBase Predicted v.2 or microT-CDS and confirmed through dual-luciferase reporter system or RNA immunoprecipitation assay (RIP). RESULTS TUG1 and CDC42 were upregulated while miR-498 was strikingly decreased in ESCC tissues and cells (P<0.0001). Besides, TUG1 suppression blocked the proliferation and invasion of ESCC cells (P<0.001). Importantly, TUG1 decrease restrained CDC42 expression via binding to miR-498 in ESCC cells. Also, the suppressive impacts of TUG1 silencing on the proliferation and invasion of ESCC cells were mitigated by miR-498 reduction. Meanwhile, the repression of proliferation and invasion induced by miR-498 elevation was weakened by CDC42 overexpression. CONCLUSIONS Inhibition of TUG1 hampered cell proliferation and invasion by downregulating CDC42 via upregulating miR-498 in ESCC cells. Thus, TUG1 might be an underlying therapeutic target for ESCC.

摘要

背景 食管鳞状细胞癌(ESCC)是一种胃肠道恶性肿瘤。牛磺酸上调基因1(TUG1)是一种长链非编码(lnc)RNA,也称为LIN00080或TI-227H,与多种疾病的肿瘤发生有关。因此,我们探究了TUG1在ESCC进展中的作用及分子机制。

材料与方法 使用定量实时聚合酶链反应(qRT-PCR)评估TUG1、微小RNA-498(miR-498)和细胞分裂周期42(CDC42)mRNA的表达模式。通过蛋白质印迹分析评估CDC42蛋白的表达水平。用细胞计数试剂盒-8(CCK-8)检测法或Transwell检测法测定细胞增殖和侵袭能力。通过在线生物信息学数据库LncBase Predicted v.2或microT-CDS预测miR-498与TUG1或CDC42之间的关系,并通过双荧光素酶报告系统或RNA免疫沉淀检测法(RIP)进行验证。

结果 ESCC组织和细胞中TUG1和CDC42上调,而miR-498显著降低(P<0.0001)。此外,抑制TUG1可阻断ESCC细胞的增殖和侵袭(P<0.001)。重要的是,在ESCC细胞中,TUG1的减少通过与miR-498结合来抑制CDC42的表达。而且,miR-498表达降低可减轻TUG1沉默对ESCC细胞增殖和侵袭的抑制作用。同时,CDC42过表达可减弱miR-498升高所诱导的增殖和侵袭抑制作用。

结论 在ESCC细胞中,抑制TUG1通过上调miR-498下调CDC42,从而阻碍细胞增殖和侵袭。因此,TUG1可能是ESCC的潜在治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6273/7077061/ac2381ea3639/medscimonit-26-e919714-g001.jpg

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