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MicroRNA-590-5p regulates cell viability, apoptosis, migration and invasion of renal cell carcinoma cell lines through targeting ARHGAP24
Molecular BioSystems Pub Date : 2017-10-11 00:00:00 , DOI: 10.1039/c7mb00406k
Lei Wang 1, 2, 3, 4 , Wan-qing Wei 4, 5, 6, 7 , Zi-yu Wu 1, 2, 3, 4 , Gong-cheng Wang 1, 4, 8, 9, 10
Affiliation  

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.

中文翻译:

MicroRNA-590-5p通过靶向ARHGAP24调节肾癌细胞的细胞活力,凋亡,迁移和侵袭

肾细胞癌(RCC)是肾恶性肿瘤死亡的主要原因。在调节癌细胞侵袭和增殖方面,MicroRNA-590-5p(miR-590-5p)在许多癌症的过程中非常重要。在我们的研究中,通过用pre-miR-590-5p(上调)或抗miR-590-5p(下调)转染来改变RCC细胞系中miR-590-5p的表达。通过流式细胞术和CCK-8分析分别测量RCC细胞系的凋亡和生存力。通过Transwell测定法估计细胞的侵袭和迁移。使用荧光素酶测定,实时PCR和蛋白质印迹测定评估了miR-590-5p与ARHGAP24表达的关联。还通过蛋白质印迹法检测凋亡和迁移相关蛋白的表达。我们发现,Caki-2和786-O细胞中的前miR-590-5p转染显示细胞活力,侵袭和迁移显着增加,并伴有细胞凋亡减少,而抗miR-590-5p转染则明显抑制与阴性对照组相比,Caki-2和786-O细胞的细胞活力,迁移和侵袭以及诱导的细胞凋亡。此外,生物信息学与荧光素酶报告基因检测相结合表明,ARHGAP24直接被miR-590-5p靶向。ARHGAP24在786-O和Caki-2细胞中的过表达显着地表达了抗miR-590-5p转染的作用,以及活性Caspase-3和Bax / Bcl-2比例的增强表达以及MMP-2和MMP的表达降低-9。
更新日期:2017-10-11
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