Circular RNA TMEM87A promotes cell proliferation and metastasis of gastric cancer by elevating ULK1 via sponging miR-142-5p
Circular RNA TMEM87A promotes cell proliferation and metastasis of gastric cancer by elevating ULK1 via sponging miR-142-5p
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环状RNA TMEM87A通过海绵miR-142-5p升高ULK1促进胃癌细胞增殖和转移
DOI：
10.1007/s00535-020-01744-1
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发表时间：
2020-11-06
影响因子：
6.3
通讯作者：
Xu, Zekuan
中科院分区：
文献类型：
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作者：
Wang, Haixiao;Sun, Guangli;Xu, Zekuan
背景环状RNA(CircRNAs)在多种癌症中是基因表达的重要调节因子。然而，CircRNAs在胃癌(GC)中的作用在很大程度上尚不清楚。因此，我们鉴定了CircTMEM87A海绵miR-142-5P通过上调ULK1的表达来促进胃癌的进展。方法通过RNA测序和实时定量聚合酶链式反应(qRT-PCR)检测CircTMEM87A在胃癌中的表达。体外和体内研究了下调或外源表达CircTMEM87A对GC细胞表型的影响。用生物信息学预测了CircTMEM87A与miRNA的相互作用，并通过RNA下拉、双荧光素酶报告基因分析和荧光原位杂交(FISH)进行了证实。应用Western印迹、GFP/MRFP-LC3斑点分析、透射电子显微镜等方法，探讨了CircTMEM87A/miR-142-5p/ULK1轴促进胃癌发生的机制。结果胃癌组织和细胞系中CircTMEM87A显著升高，其高表达与胃癌患者预后不良密切相关。在体外，CircTMEM87A基因的敲除抑制了细胞的生长、迁移、侵袭和诱导细胞凋亡，在体内也抑制了胃癌的致瘤性和肺转移潜能。而约TMEM87A过表达则有相反的作用。此外，我们还证实了CircTMEM87A可以作为miR-142-5P的海绵，调节ULK1的表达和胃癌的进展。结论我们的研究结果表明，CircTMEM87A通过miR-142-5p/ULK1轴发挥癌基因的作用。CircTMEM87A可能是一种预测预后的生物标志物，也是一种有前途的治疗靶点。
BackgroundCircular RNAs (circRNAs) act as vital regulators of gene expression in a variety of cancers. However, the role of circRNAs in gastric cancer (GC) remains largely unexplored. Herein, we identified that circTMEM87A sponges miR-142-5p to promote GC progression through up-regulating ULK1 expression.MethodsThe expression of circTMEM87A in GC was determined by RNA sequencing and quantitative real-time PCR (qRT-PCR). The effects of knockdown or exogenous expression of circTMEM87A on GC cell phenotypes were evaluated both in vitro and in vivo. The interacting miRNA of circTMEM87A was predicted by bioinformatics and confirmed by RNA pull-down, dual-luciferase reporter assay and fluorescence in situ hybridization (FISH). The mechanism by which circTMEM87A/miR-142-5p/ULK1 axis promotes GC was determined by western blot, GFP/mRFP-LC3 puncta analysis, transmission electron microscope (TEM).ResultsCircTMEM87A was dramatically elevated in GC tissues and cell lines, and high circTMEM87A expression was closely correlated with poor prognosis of GC patients. Knockdown of circTMEM87A suppressed cell growth, migration, invasion and induced apoptosis in vitro, as well as inhibited GC tumorigenicity and lung metastasis potential in vivo. Meanwhile, circTMEM87A overexpression had the opposite effects. Furthermore, we demonstrated that circTMEM87A could act as a sponge of miR-142-5p to regulate ULK1 expression and GC progression.ConclusionsOur findings suggest that circTMEM87A functions as an oncogene through the miR-142-5p/ULK1 axis in GC. CircTMEM87A might be a prognostic biomarker as well as a promising therapeutic target for GC.