microRNA-199a-3p functions as tumor suppressor by regulating glucose metabolism in testicular germ cell tumors
microRNA-199a-3p functions as tumor suppressor by regulating glucose metabolism in testicular germ cell tumors
复制标题
microRNA-199a-3p 通过调节睾丸生殖细胞肿瘤中的葡萄糖代谢发挥肿瘤抑制剂的作用
DOI：
10.3892/mmr.2016.5472
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发表时间：
2016-09-01
影响因子：
3.4
通讯作者：
Li, Dan
中科院分区：
文献类型：
--
作者：
Liu, Xiaowen;Duan, Hongyan;Li, Dan
microRNA（miR）-199a-3p在癌症的发生和发展中起着关键作用。为了进一步了解miR-199 a-3 p在睾丸肿瘤中的作用机制，本研究探讨了miR-199 a-3 p对有氧糖酵解的调控及其对代谢的影响。使用3- 4，5-二甲基噻唑-2-基-2，5-二苯基四唑溴化物、伤口愈合和流式细胞术测定，确定miR-199 a-3 p在Ntera-2细胞中的过表达引起细胞生长和迁移的抑制。进一步的生物化学方法和代谢基因的高通量定量聚合酶链反应阵列显示，抑制miR-199 a-3 p显著提高了Ntera-2细胞的乳酸产生，并且在miR-199 a-3 p处理后鉴定了12个差异表达基因，包括2个上调基因和10个下调基因。在临床样本中，乳酸脱氢酶A、单羧酸转运蛋白1、磷酸甘油酸激酶1和TP 53诱导的糖酵解和凋亡调节因子4个选定基因在恶性睾丸生殖细胞肿瘤中显著过表达，且其表达与miR-199 a-3 p的表达呈负相关，提示这4个基因可能受到miR-199 a-3 p的影响。利用生物信息学分析，确定转录因子Sp1结合位点的启动子区的四个选定的基因。此外，预测miR-199 a-3 p与Sp1 mRNA的3-非翻译区中的保守靶序列结合，表明miR-199 a-3 p可能通过Sp1下调这四个代谢基因。研究表明，miR-199 a-3 p的异常表达和激活可能在睾丸癌患者有氧糖酵解和肿瘤发生中发挥重要作用。因此，miR-199 a-3 p可能是睾丸肿瘤预后和治疗的潜在生物标志物。
microRNA (miR)-199a-3p serves critical roles in cancer development and progression. In order to improve knowledge of the functional mechanism of miR-199a-3p in testicular tumors, the present study characterized the regulation of aerobic glycolysis by miR-199a-3p and its impact on metabolism. Using 3-4,5-dimethylthiazol-2-yl-2,5 diphenyl tetrazolium bromide, wound healing and flow cytometry assays, it was determined that overexpression of miR-199a-3p in Ntera-2 cells caused suppression of cell growth and migration. Further biochemical methods and high-throughput quantitative polymerase chain reaction array of metabolic genes showed that inhibition of miR-199a-3p markedly elevated lactate production and 12 differentially expressed genes, including 2 upregulated and 10 downregulated genes, were identified following treatment with miR-199a-3p in Ntera-2 cells. In clinical samples, four selected genes, lactate dehydrogenase A, monocarboxylate transporter 1, phosphoglycerate kinase 1 and TP53-inducible glycolysis and apoptosis regulator, were significantly overexpressed in malignant testicular germ cell tumor, and their expression inversely correlated with the expression of miR-199a-3p, suggesting that these four genes may be affected by miR-199a-3p. Using bioinformatics analysis, the transcription factor Sp1 binding site was identified in the promoter region of the four selected genes. In addition, miR-199a-3p was predicted to bind to conservative target sequences in the 3-untranslated region of Sp1 mRNA, suggesting that miR-199a-3p may downregulate these four metabolic genes through Sp1. It was demonstrated the dysregulated expression and activation of miR-199a-3p may serve important roles in aerobic glycolysis and tumorigenesis in patients with testicular cancer. Therefore, miR-199a-3p may be a potential biomarker in the prognosis and treatment of testicular tumors.