MicroRNA-184 Modulates Doxorubicin Resistance in Osteosarcoma Cells by Targeting BCL2L1.
MicroRNA-184 Modulates Doxorubicin Resistance in Osteosarcoma Cells by Targeting BCL2L1.
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DOI：
10.12659/msm.896451
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发表时间：
2016-05-25
期刊：
影响因子：
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通讯作者：
Shi FJ
中科院分区：
文献类型：
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作者：
Lin BC;Huang D;Yu CQ;Mou Y;Liu YH;Zhang DW;Shi FJ
骨肉瘤的早期转移是高致死性的，并且对药物和放射治疗反应差。MicroRNA（miRNAs）是一类在转录后水平调节基因表达的小分子非编码RNA。然而，具体的miRNAs的详细功能并不完全清楚。本研究的目的是探讨miR-184作为人骨肉瘤耐药介质的作用。采用qRT-PCR方法分析阿霉素处理后的OS细胞系U-2 OS和MG-63中miR-184的表达水平。将miR-184 agomir或miR-184 iagomir转移到细胞中以调节miR-184。通过TargetScan预测miR-184的靶点，并通过荧光素酶报告基因检测进行验证。Western blot检测Bcl-2样蛋白1（BCL 2L 1）表达。Annexin V染色检测细胞凋亡，流式细胞仪检测细胞凋亡率。阿霉素诱导OS细胞系U-2 OS和MG-63中miR-184的时间依赖性表达。荧光素酶报告基因分析确定BCL 2L 1为miR-184的直接靶基因。此外，多柔比星降低了BCL 2L 1的表达，这被miR-184过表达逆转，并且在OS细胞中被miR-184抑制进一步降低。此外，miR-184 agomir减少了阿霉素诱导的细胞凋亡，而miR-184 apomir增强了OS细胞的凋亡，这表明miR-184的上调有助于OS细胞系的化学抗性。我们的数据显示，miR-184在接受阿霉素治疗的OS患者中上调，并导致对靶向BCL 2L 1的药物治疗反应不良。
Early metastasis of osteosarcoma (OS) is highly lethal and responds poorly to drug and radiation therapies. MicroRNAs (miRNAs) are a class of small noncoding RNAs that modulate gene expression at the post-transcriptional level. However, the detailed functions of specific miRNAs are not entirely understood. The aim of the present study was to investigate the role of miR-184 as a mediator of drug resistance in human osteosarcoma. qRT-PCR was used to analyze the expression level of miR-184 in OS cell line U-2 OS and MG-63 treated with doxorubicin. MiR-184 agomir or miR-184 antagomir was transferred into cells to regulated miR-184. The target of miR-184 was predicted by TargetScan and confirmed by luciferase reporter assay. Bcl-2-like protein 1 (BCL2L1) expression was detected by Western blot. Cell apoptosis was determined by Annexin V staining and analysis by flow cytometry. Doxorubicin induced time-dependent expression of miR-184 in OS cell line U-2 OS and MG-63. Luciferase reporter assay identified BCL2L1 as the direct target gene of miR-184. Furthermore, doxorubicin reduced BCL2L1 expression, which was reversed by miR-184 overexpression and further decreased by miR-184 inhibition in OS cells. In addition, miR-184 agomir reduced doxorubicin-induced cell apoptosis, whereas miR-184 antagomir enhanced apoptosis in OS cells, suggesting that up-regulation of miR-184 contributes to chemoresistance of the OS cell line. Our data show that miR-184 was up-regulated in OS patients treated with doxorubicin therapy and leads to poor response to drug therapy by targeting BCL2L1.