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环状 PRMT5 通过靶向 miR-188-5p/HK2 轴促进肝癌细胞的增殖、迁移和糖酵解。

Circ-PRMT5 enhances the proliferation, migration and glycolysis of hepatoma cells by targeting miR-188-5p/HK2 axis.

机构信息

Department of General surgery, Xiangyang NO. 1 People's Hospital, Affiliated Hospital of Hubei University of Medicine, Xiangyang, Hubbei, China.

Department of General surgery, Xiangyang NO. 1 People's Hospital, Affiliated Hospital of Hubei University of Medicine, Xiangyang, Hubbei, China.

出版信息

Ann Hepatol. 2020 May-Jun;19(3):269-279. doi: 10.1016/j.aohep.2020.01.002. Epub 2020 Jan 27.

Abstract

INTRODUCTION AND OBJECTIVES

Circular RNA (circRNA) has been demonstrated as a critical regulator in human cancer, including hepatocellular carcinoma (HCC). Nevertheless, the role of circ-PRMT5 in HCC remains largely unknown.

PATIENTS OR MATERIALS AND METHODS

The real-time quantitative polymerase chain reaction (RT-qPCR) was performed to assess the expression levels of circ-PRMT5, miR-188-5p and anti-Hexokinase II (HK2) in HCC tissues and cells. The cell proliferation, migration and glycolysis were determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazol-3-ium bromide (MTT), transwell migration assay, and indicated kits, respectively. The interaction relationship between miR-188-5p and circ-PRMT5 or HK2 was analyzed by the bioinformatics database, dual-luciferase reporter assay, and RNA immunoprecipitation (RIP) assay. The western blot assay was used to analyze the expression level of HK2. The functional role of circ-PRMT5 in vivo was assessed by a xenograft experiment.

RESULTS

Circ-PRMT5 was elevated in HCC tissues and cells than matched control groups. Furthermore, loss-of-functional experiments revealed that the silencing of circ-PRMT5 could repress proliferation, migration, glycolysis in vitro and tumor growth in vivo. Moreover, we also confirmed that overexpression of circ-PRMT5 abolished the effects on HCC cells induced by upregulating miR-188-5p. In addition, overexpression of miR-188-5p could repress the development of HCC. More importantly, HK2 was a target gene of miR-188-5p, and miR-188-5p regulated proliferation, migration, glycolysis of HCC cells by specifically binding to HK2. Mechanistically, circ-PRMT5 could act as a sponge of miR-188-5p to regulate the expression of HK2.

CONCLUSION

In summary, circ-PRMT5 might play a key role in proliferation, migration, glycolysis of HCC cells via miR-188-5p/HK2 axis, which indicated that circ-PRMT5 might be a potential therapeutic target for HCC treatment.

摘要

简介和目的

环状 RNA(circRNA)已被证明是人类癌症(包括肝细胞癌(HCC))的关键调控因子。然而,circ-PRMT5 在 HCC 中的作用在很大程度上仍然未知。

患者或材料和方法

实时定量聚合酶链反应(RT-qPCR)用于评估 HCC 组织和细胞中 circ-PRMT5、miR-188-5p 和抗己糖激酶 II(HK2)的表达水平。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑-3-溴化铵(MTT)、Transwell 迁移实验和指示试剂盒分别确定细胞增殖、迁移和糖酵解。通过生物信息学数据库、双荧光素酶报告基因实验和 RNA 免疫沉淀(RIP)实验分析 miR-188-5p 与 circ-PRMT5 或 HK2 的相互作用关系。Western blot 实验用于分析 HK2 的表达水平。通过异种移植实验评估 circ-PRMT5 在体内的功能作用。

结果

circ-PRMT5 在 HCC 组织和细胞中的表达高于匹配对照组。此外,功能丧失实验表明,circ-PRMT5 的沉默可以抑制体外增殖、迁移和糖酵解以及体内肿瘤生长。此外,我们还证实,过表达 circ-PRMT5 可以消除上调 miR-188-5p 对 HCC 细胞的影响。此外,miR-188-5p 的过表达可以抑制 HCC 的发展。更重要的是,HK2 是 miR-188-5p 的靶基因,miR-188-5p 通过特异性结合 HK2 调节 HCC 细胞的增殖、迁移和糖酵解。机制上,circ-PRMT5 可以作为 miR-188-5p 的海绵来调节 HK2 的表达。

结论

总之,circ-PRMT5 可能通过 miR-188-5p/HK2 轴在 HCC 细胞的增殖、迁移和糖酵解中发挥关键作用,表明 circ-PRMT5 可能是 HCC 治疗的潜在治疗靶点。

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