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Circ_0002711 敲低通过调节 miR-1244/ROCK1 轴抑制卵巢癌细胞生长和有氧糖酵解。

Circ_0002711 knockdown suppresses cell growth and aerobic glycolysis by modulating miR-1244/ROCK1 axis in ovarian cancer.

机构信息

Department of Obstetrics and Gynaecology, Central Hospital of Enshi Tujia and Miao Autonomous Prefecture, Enshi City, Hubei Province, China.

出版信息

J Biosci. 2021;46.

Abstract

It has been well investigated that circular RNAs (circRNAs) play important roles in various cancers. The function of circ_0002711 and its underlying mechanisms in ovarian cancer (OC) remain unknown. qRT-PCR and western blot were performed to detect the expressions of circ_0002711, microRNA-1244 (miR-1244), and Rho kinase 1 (ROCK1) in OC tissues and cells. MTT assay and colony formation assay were employed to evaluate cell proliferation. Detection of lactate production, glucose uptake, and ATP level and oxygen consumption were used to determine Warburg effect. Western blot was used to examine glycolysis or proliferationrelated genes. Dual-luciferase reporter assay and RIP pull down assay were used to address the relationship among circ_0002711, miR-1244, and ROCK1. tumor growth was evaluated in nude mice. Circ_0002711 was upregulated in OC tissues and cell lines. Circ_0002711 downregulation inhibited cell viability, colony formation ability and aerobic glycolysis. Circ_0002711 contained binding sites with miR1244. Moreover, loss of miR-1244 undermined circ_0002711 downregulation-mediated function. ROCK1 contained binding sites with miR-1244. MiR-1244 upregulation suppressed cell proliferation and aerobic glycolysis, which was rescued by enhanced expression of ROCK1. Circ_0002711 knockdown hampered ROCK1 expression by upregulating miR-1244 expression. Finally, decreased expression of circ_0002711 inhibited tumor growth . Circ_0002711/miR-1244/ROCK1 axis regulated Warburg effect and tumor growth .

摘要

已经有充分的研究表明,环状 RNA(circRNAs)在各种癌症中发挥着重要作用。circ_0002711 在卵巢癌(OC)中的作用及其潜在机制尚不清楚。qRT-PCR 和 Western blot 用于检测 OC 组织和细胞中 circ_0002711、microRNA-1244(miR-1244)和 Rho 激酶 1(ROCK1)的表达。MTT assay 和集落形成 assay 用于评估细胞增殖。检测乳酸生成、葡萄糖摄取和 ATP 水平以及耗氧量用于确定瓦博格效应。Western blot 用于检测糖酵解或增殖相关基因。双荧光素酶报告 assay 和 RIP pull down assay 用于研究 circ_0002711、miR-1244 和 ROCK1 之间的关系。裸鼠体内肿瘤生长情况进行评估。circ_0002711 在 OC 组织和细胞系中上调。circ_0002711 下调抑制细胞活力、集落形成能力和有氧糖酵解。circ_0002711 含有与 miR1244 结合的位点。此外,miR-1244 的缺失破坏了 circ_0002711 下调介导的功能。ROCK1 含有与 miR-1244 结合的位点。miR-1244 上调抑制细胞增殖和有氧糖酵解,这可以通过增强 ROCK1 的表达来挽救。circ_0002711 下调通过上调 miR-1244 表达来抑制 ROCK1 表达。最后,circ_0002711 的表达降低抑制了肿瘤的生长。circ_0002711/miR-1244/ROCK1 轴调节瓦博格效应和肿瘤生长。

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