Zhao Yue, Xu Kaicheng, Liu Peng
Department of Vascular Surgery, China-Japan Union Hospital, Jilin University, Changchun, China.
Department of Anesthesiology, China-Japan Union Hospital, Jilin University, Changchun, China.
Cell Physiol Biochem. 2018;45(4):1581-1589. doi: 10.1159/000487719. Epub 2018 Feb 21.
BACKGROUND/AIMS: MicroRNAs (miRNAs) play an essential role in the tumorigenesis of osteosarcoma (OS). However, the effects of miR-1248 on chemo-resistant potential of OS have not been studied. Here, we addressed this question.
The levels of miR-1248 and apoptotic protein angiotensin II type 1 receptor (AGTR1) in OS specimens were examined by RT-qPCR and Western blotting, respectively. The relationship between miR-1248 and AGTR1 was determined by analysis of Spearman's Rank Correlation Coefficients. The patient survival was determined with Kaplan-Meier curves. Bioinformatics analyses were done to predict microRNAs (miRNAs) that target AGTR1. The functional binding of miRNAs to AGTR1 mRNA was examined by a dual luciferase reporter assay. Cell viability was determined by an CCK-8 assay. Apoptosis was determined by a fluorescence-based apoptosis assay.
The levels of miR-1248 were significantly elevated while the levels of AGTR1 were significantly decreased in OS specimens than in paired adjacent normal tissue. The levels of miR-1248 were negatively correlated to the levels of AGTR1. Moreover, the patients with high miR-1248 levels had poorer survival than those with low MiR-1248 levels, and the patients with low AGTR1 levels had poorer survival than those with high AGTR1 levels. MiR-1248 inhibited protein translation of AGTR1, through binding to the 3'-UTR of the AGTR1 mRNA. The AGTR1-mediated cell apoptosis was suppressed by overexpressing miR-1248, and was augmented by depleting miR-1248.
Increased miR-1248 expression in OS may inhibit AGTR1-mediated cancer cell death in chemotherapy. The outcome of chemotherapy may be improved by the suppression of miR-1248 in OS cells.
背景/目的：微小RNA（miRNA）在骨肉瘤（OS）的肿瘤发生中起重要作用。然而，miR-1248对OS化疗耐药潜能的影响尚未得到研究。在此，我们探讨了这个问题。
分别通过逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测OS标本中miR-1248和凋亡蛋白血管紧张素II 1型受体（AGTR1）的水平。通过分析Spearman等级相关系数确定miR-1248与AGTR1之间的关系。用Kaplan-Meier曲线确定患者生存率。进行生物信息学分析以预测靶向AGTR1的微小RNA（miRNA）。通过双荧光素酶报告基因检测法检测miRNA与AGTR1 mRNA的功能性结合。通过细胞计数试剂盒-8（CCK-8）检测法测定细胞活力。通过基于荧光的凋亡检测法测定细胞凋亡。
与配对的相邻正常组织相比，OS标本中miR-1248水平显著升高，而AGTR1水平显著降低。miR-1248水平与AGTR1水平呈负相关。此外，miR-1248水平高的患者生存率低于miR-1248水平低的患者，AGTR1水平低的患者生存率低于AGTR1水平高的患者。miR-1248通过与AGTR1 mRNA的3'-非翻译区（3'-UTR）结合抑制AGTR1的蛋白质翻译。过表达miR-1248可抑制AGTR1介导的细胞凋亡，而敲低miR-1248则可增强细胞凋亡。
OS中miR-1248表达增加可能抑制化疗中AGTR1介导的癌细胞死亡。抑制OS细胞中的miR-1248可能改善化疗结果。
