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MicroRNA-21 Inhibits the Apoptosis of Osteosarcoma Cell Line SAOS-2 via Targeting Caspase 8

机译:MicroRNA-21通过靶向Caspase 8抑制骨肉瘤细胞系Saos-2的凋亡

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摘要

Currently, multiple microRNAs (miRNAs) have been found to play vital roles in the pathogenesis of osteosarcoma. This study aimed to investigate the role of miR-21 in osteosarcoma. The level of miR-21 in 20 pairs of osteosarcoma and corresponding adjacent tissues was monitored by qPCR. Human osteosarcoma cell line SAOS-2 was transfected with either miR-21 mimic or miR-21 inhibitor, and then cell viability, survival, and apoptosis were measured by MTT, colony formation assay, and flow cytometry. A target of miR-21 was predicted by the microRNA.org database and verified in vitro by using luciferase reporter, qPCR, and Western blot analyses. Finally, cells were cotransfected with siRNA against caspase 8 and miR-21 inhibitor, and the apoptotic cell rate was determined again. Results showed that the mRNA level of miR-21 was highly expressed in osteosarcoma tissues compared with adjacent tissues. Overexpression of miR-21 improved cell viability and survival but suppressed apoptosis. Caspase 8 was a direct target of miR-21, and it was negatively regulated by miR-21. Moreover, miR-21 suppression attenuated caspase 8 silencing and induced the decrease in apoptosis. In conclusion, overexpression of miR-21 suppressed SAOS-2 cell apoptosis via directly targeting caspase 8.
机译:目前,已发现多个MicroRNA(miRNA)在骨肉瘤的发病机制中起到重要的作用。本研究旨在探讨miR-21在骨肉瘤中的作用。通过QPCR监测20对骨肉瘤和相应的相邻组织中miR-21的水平。用miR-21模拟或miR-21抑制剂转染人骨肉瘤细胞系SAOS-2,然后通过MTT,菌落形成测定和流式细胞术测量细胞活力,存活和细胞凋亡。 MICRORNA.org数据库预测MIR-21的靶标,并通过使用荧光素酶报告,QPCR和Western Blot分析进行体外验证。最后,将细胞用针对半胱天冬酶8和miR-21抑制剂的siRNA分配,并且再次测定凋亡细胞率。结果表明,与相邻组织相比,在骨肉瘤组织中,MIR-21的mRNA水平高度表达。 miR-21的过度表达改善了细胞活力和存活但抑制了细胞凋亡。胱天蛋白酶8是miR-21的直接靶标,并通过miR-21负调节。此外,miR-21抑制减毒胱天蛋白酶8沉默并诱导细胞凋亡的降低。总之,MiR-21的过度表达通过直接靶向Caspase 8抑制Saos-2细胞凋亡。

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