MicroRNA-590-5p通过靶向ARHGAP24调控肾细胞癌细胞系的细胞活力、凋亡、迁移和侵袭

IF 3.743 Q2 Biochemistry, Genetics and Molecular Biology Molecular BioSystems Pub Date : 2017-10-11 DOI:10.1039/C7MB00406K
Lei Wang, Wan-qing Wei, Zi-yu Wu and Gong-cheng Wang
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引用次数: 17

摘要

肾细胞癌(RCC)是肾脏恶性肿瘤死亡的主要原因。MicroRNA-590-5p (miR-590-5p)在许多癌症的过程中调控癌细胞的侵袭和增殖具有重要意义。在我们的研究中,通过转染pre-miR-590-5p(上调)或anti-miR-590-5p(下调)来改变miR-590-5p在RCC细胞系中的表达。流式细胞术和CCK-8分析分别检测细胞凋亡和细胞活力。Transwell法测定细胞的侵袭和迁移。采用荧光素酶测定、实时PCR和western blot法评估miR-590-5p与ARHGAP24表达的关系。western blotting检测细胞凋亡和迁移相关蛋白的表达。我们发现,转染pre-miR-590-5p后,Caki-2和786-O细胞的细胞活力、侵袭和迁移显著增加,同时细胞凋亡减少,而转染anti-miR-590-5p后,与阴性对照组相比,Caki-2和786-O细胞的细胞活力、迁移和侵袭明显受到抑制,并诱导细胞凋亡。此外,生物信息学结合荧光素酶报告基因检测表明,ARHGAP24是miR-590-5p的直接靶点。ARHGAP24在786-O和Caki-2细胞中的过表达表现出抗mir -590-5p转染的作用,同时活性Caspase-3和Bax/Bcl-2比值的表达增强,MMP-2和MMP-9的表达降低。这些发现表明,miR-590-5p/ARHGAP24似乎是RCC治疗的潜在有益靶点。
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MicroRNA-590-5p regulates cell viability, apoptosis, migration and invasion of renal cell carcinoma cell lines through targeting ARHGAP24

Renal cell carcinoma (RCC) is the leading cause of death in renal malignancies. MicroRNA-590-5p (miR-590-5p) is of great importance in the processes of many cancers regarding regulation of cancer cell invasion and proliferation. In our study, alternation of miR-590-5p expression in RCC cell lines through transfection with pre-miR-590-5p (up-regulation) or anti-miR-590-5p (down-regulation) was performed. Apoptosis and viability of RCC cell lines were measured by flow cytometry and CCK-8 analysis, respectively. Cell invasion and migration were estimated by Transwell assay. The association of miR-590-5p with ARHGAP24 expression was evaluated using luciferase assays, real-time PCR and western blot assay. The expressions of apoptosis and migration-related protein were also measured by western blotting. We found that pre-miR-590-5p transfection in Caki-2 and 786-O cells showed significant increases in cell viability, invasion and migration, which were accompanied by decreased cell apoptosis, while anti-miR-590-5p transfection obviously inhibited the cell viability, migration and invasion of Caki-2 and 786-O cells as well as induced apoptosis, compared with the negative control group. Furthermore, bioinformatics combined with luciferase reporter assays indicated that ARHGAP24 is directly targeted by miR-590-5p. ARHGAP24 overexpression in 786-O and Caki-2 cells phenocopied the effects of anti-miR-590-5p transfection along with enhanced expression of active Caspase-3 and Bax/Bcl-2 ratio as well as decreased expression of MMP-2 and MMP-9. These findings suggested that miR-590-5p/ARHGAP24 seems to function as a potentially beneficial target for RCC treatment.

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来源期刊
Molecular BioSystems
Molecular BioSystems 生物-生化与分子生物学
CiteScore
2.94
自引率
0.00%
发文量
0
审稿时长
2.6 months
期刊介绍: Molecular Omics publishes molecular level experimental and bioinformatics research in the -omics sciences, including genomics, proteomics, transcriptomics and metabolomics. We will also welcome multidisciplinary papers presenting studies combining different types of omics, or the interface of omics and other fields such as systems biology or chemical biology.
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