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环状RNA C6orf132通过作用于miR-873-5p/PRKAA1轴促进缺氧条件下胃癌细胞的增殖、迁移、侵袭和糖酵解。

CircC6orf132 Facilitates Proliferation, Migration, Invasion, and Glycolysis of Gastric Cancer Cells Under Hypoxia by Acting on the miR-873-5p/PRKAA1 Axis.

作者信息

Chen Weizhi, Ji Yanhong

机构信息

Department of Pathogenic Biology and Immunology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an, China.

Department of Radiology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China.

出版信息

Front Genet. 2021 Sep 30;12:636392. doi: 10.3389/fgene.2021.636392. eCollection 2021.

Abstract

Hypoxia is a crucial factor in the progression of various tumors, including gastric cancer (GC). Circular RNAs (circRNAs) are important regulators in GC, and this study focused on researching circC6orf132 in GC progression under hypoxia. experiments were performed in GC cells under hypoxia (1% O). CircC6orf132, microRNA-873-5p (miR-873-5p), and protein kinase AMP-activated alpha 1 catalytic subunit (PRKAA1) levels were examined by real-time polymerase chain reaction (qRT-PCR). Colony formation assay and transwell assay were used for detecting cell proliferation and migration or invasion. Glycolytic metabolism was evaluated using lactate production, glucose uptake, and adenosine triphosphate (ATP) level and extracellular acidification rate (ECAR). Western blotting was performed for determining protein expression. The target interaction was analyzed by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. assay was conducted mouse xenograft model. The expression of circC6orf132 was significantly high in GC cells under hypoxia. Hypoxia-induced GC proliferation, migration, invasion, and glycolysis were reversed by silencing circC6orf132. CircC6orf132 targeted miR-873-5p; and the inhibition of circC6orf132 knockdown for the effects of hypoxia on GC cells was abrogated by miR-873-5p inhibitor. PRKAA1 was validated as a downstream gene of miR-873-5p, and miR-873-5p functioned as an anticancer molecule in GC cells under hypoxia by downregulating PRKAA1 level. CircC6orf132 could regulate PRKAA1 by sponging miR-873-5p. CircC6orf132/miR-873-5p/PRKAA1 axis could regulate GC progression under the hypoxic condition. CircC6orf132 downregulation reduced tumorigenesis through affecting the miR-873-5p/PRKAA1 axis. CircC6orf132 has been affirmed to promote proliferation, migration, invasion, and glycolysis in GC under hypoxia, partly by depending on the regulation of miR-873-5p/PRKAA1 axis.

摘要

缺氧是包括胃癌(GC)在内的各种肿瘤进展的关键因素。环状RNA(circRNA)是GC中的重要调节因子,本研究聚焦于研究缺氧条件下circC6orf132在GC进展中的作用。在缺氧(1%氧气)条件下对GC细胞进行实验。通过实时聚合酶链反应(qRT-PCR)检测circC6orf132、微小RNA-873-5p(miR-873-5p)和蛋白激酶AMP激活的α1催化亚基(PRKAA1)的水平。采用集落形成试验和Transwell试验检测细胞增殖、迁移或侵袭能力。通过检测乳酸生成、葡萄糖摄取、三磷酸腺苷(ATP)水平和细胞外酸化率(ECAR)评估糖酵解代谢。采用蛋白质印迹法测定蛋白质表达。通过双荧光素酶报告基因和RNA免疫沉淀(RIP)试验分析靶标相互作用。在小鼠异种移植模型上进行试验。缺氧条件下GC细胞中circC6orf132的表达显著升高。沉默circC6orf132可逆转缺氧诱导的GC增殖、迁移、侵袭和糖酵解。circC6orf132靶向miR-873-5p;miR-873-5p抑制剂可消除circC6orf132敲低对缺氧对GC细胞影响的抑制作用。PRKAA1被证实为miR-873-5p的下游基因,在缺氧条件下,miR-873-5p通过下调PRKAA1水平在GC细胞中发挥抗癌分子的作用。circC6orf132可通过海绵吸附miR-873-5p来调节PRKAA1。circC6orf132/miR-873-5p/PRKAA1轴可在缺氧条件下调节GC进展。circC6orf132的下调通过影响miR-873-5p/PRKAA1轴降低肿瘤发生。circC6orf132已被证实可在缺氧条件下促进GC的增殖、迁移、侵袭和糖酵解,部分依赖于对miR-873-5p/PRKAA1轴的调节。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/897e/8514671/8c29d17051f2/fgene-12-636392-g001.jpg

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