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JHDM1D-AS1 aggravates the development of gastric cancer through miR-450a-2-3p-PRAF2 axis
Life Sciences ( IF 5.2 ) Pub Date : 2020-11-24 , DOI: 10.1016/j.lfs.2020.118805
Min Wu , Yi Liu , Yan-Song Pu , Yu Ma , Jian-Hua Wang , En-Qi Liu

Aims

To investigate the molecular function and mechanisms of JHDM1D antisense 1 (JHDM1D-AS1) during gastric cancer (GC) progression.

Materials and methods

The qPCR assay was used to detect the JHDM1D-AS1 and miR-450a-2-3p expression levels in GC tissues and cell lines. Bioinformatics analysis was used for exploring the lncRNA-microRNA-mRNA interaction network. We performed dual-luciferase reporter assay and qPCR assay in order to validate the direct interactions. We explored the JHDM1D-AS1 and miR-450a-2-3p on GC progression by using JHDM1D-AS1 siRNA and miR-450a-2-3p inhibitor; in vitro CCK-8 assay, colony formation assay, and invasion assay were conducted. Further, in vivo animal experiments were performed, and the expression levels of miR-450a-2-3p and PRAF2 in the tumor tissues were detected using qPCR and western blot analysis.

Key findings

The expression levels of JHDM1D-AS1 and miR-450a-2-3p in GC tissues and cell lines were higher and lower as compared to those in the corresponding normal controls, respectively. Moreover, high levels of JHDM1D-AS1 were closely related with metastasis and the GC TNM stage. Functionally, JHDM1D-AS1 depletion caused an obvious reduction in cell proliferation and invasion both in vitro and in vivo, while the addition of miR-450a-2-3p inhibitor could nullify these effects. Mechanically, JHDM1D-AS1 promoted GC progression via the sponging of miR-450a-2-3p in order to increase PRAF2 expression.

Significance

The present results showed that the increased expression of JHDM1D-AS1 was closely associated with tumor progression of GC. JHDM1D-AS1/miR-450a-2-3p/PRAF2 axis may be a promising target for GC treatment.



中文翻译:

JHDM1D-AS1通过miR-450a-2-3p-PRAF2轴加重了胃癌的发展

目的

调查JHDM1D反义1(JHDM1D-AS1)在胃癌(GC)进展过程中的分子功能和机制。

材料和方法

qPCR分析用于检测GC组织和细胞系中的JHDM1D-AS1和miR-450a-2-3p表达水平。生物信息学分析用于探索lncRNA-microRNA-mRNA相互作用网络。为了验证直接的相互作用,我们进行了双重荧光素酶报告基因测定和qPCR测定。我们通过使用JHDM1D-AS1 siRNA和miR-450a-2-3p抑制剂探索了JHDM1D-AS1和miR-450a-2-3p对GC进程的影响;进行了体外CCK-8测定,菌落形成测定和侵袭测定。此外,进行了体内动物实验,并使用qPCR和蛋白质印迹分析检测了肿瘤组织中miR-450a-2-3p和PRAF2的表达水平。

主要发现

与相应的正常对照组相比,JHDM1D-AS1和miR-450a-2-3p在GC组织和细胞系中的表达水平分别更高和更低。此外,高水平的JHDM1D-AS1与转移和GC TNM分期密切相关。从功能上讲,JHDM1D-AS1耗竭在体内外均引起细胞增殖和侵袭的明显减少,而添加miR-450a-2-3p抑制剂则可消除这些作用。在机械上,JHDM1D-AS1通过绵延miR-450a-2-3p促进GC进程,以增加PRAF2的表达。

意义

目前的结果表明,JHDM1D-AS1的表达增加与GC的肿瘤进展密切相关。JHDM1D-AS1 / miR-450a-2-3p / PRAF2轴可能是有希望的GC治疗靶标。

更新日期:2020-12-06
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