Comprehensive gene and microRNA expression profiling reveals a role for miRNAs in the oncogenic roles of SphK1 in papillary thyroid cancer
Comprehensive gene and microRNA expression profiling reveals a role for miRNAs in the oncogenic roles of SphK1 in papillary thyroid cancer
复制标题
全面的基因和 microRNA 表达谱揭示了 miRNA 在 SphK1 在甲状腺乳头状癌中的致癌作用中的作用
DOI：
10.1007/s00432-016-2315-0
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发表时间：
2017-04-01
影响因子：
3.6
通讯作者：
Guan, Hongyu
中科院分区：
文献类型：
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作者：
Liang, Weiwei;Xie, Zhiwei;Guan, Hongyu
目的鞘氨醇激酶1(SphK1)在包括甲状腺癌在内的多种癌症中的致癌作用已被充分证明。然而，与SphK1致癌作用相关的microRNAs(MiRNAs)在很大程度上仍不清楚。方法分别用数字基因表达分析(DGE)和小RNA序列分析方法分析了TPC1载体和TPC1 SphK1细胞中的全局基因和miRNA的表达。用microT-CDS研究miRNA-mRNA的相互作用，并用CytoScape®可视化预测的网络。通过进行Transwell侵袭和伤口愈合试验评估细胞的侵袭和迁移。用荧光素酶报告基因和免疫印迹法检测miR-144-3对纤维连接蛋白1(FN1)的靶向性。结果在本研究中，我们发现SphK1的过表达对甲状腺乳头状癌(PTC)TPC1细胞中46个miRNAs和506个mRNAs的表达具有差异性调节作用。结合对信使核糖核酸靶标的生物信息学预测和关于信使核糖核酸表达的DGE数据，使我们能够识别解除调控的miRNAs的信使核糖核酸靶标。实验证实了miR-144-3p与FN1之间的直接相互作用，介导了SphK1在PTC细胞中的促侵袭作用。结论SphK1的过表达驱动了一个调控PTC细胞miRNA和mRNA表达的网络。我们还展示了miR-144-3p和FN1在介导SphK1的致癌功能中所起的作用，从而加深了对PTC病因的理解。
PurposeThe oncogenic roles of sphingosine kinase 1 (SphK1) in various cancers, including thyroid cancer, have been well demonstrated. However, the microRNAs (miRNAs) associated with the oncogenic roles of SphK1 remain largely unknown.MethodsGlobal gene and miRNA expression in TPC1-Vector and TPC1-SphK1 cells was analyzed using digital gene expression (DGE) analysis and small RNA-seq, respectively. miRNA–mRNA interactions were explored by microT-CDS, and the predicted networks were visualized using CytoScape®. Cell invasion and migration were assessed by performing Transwell invasion and wound-healing assays. Luciferase reporter and immunoblot assays were used to evaluate the targeting of fibronectin 1 (FN1) by miR-144-3p.ResultsIn this study, we found that overexpression of SphK1 differentially regulates the expression of 46 miRNAs and 506 mRNAs in papillary thyroid cancer (PTC) TPC1 cells. Combining bioinformatics predictions of mRNA targets with DGE data on mRNA expression allowed us to identify the mRNA targets of deregulated miRNAs. The direct interaction between miR-144-3p and FN1, which mediates the pro-invasive role of SphK1 in PTC cells, was experimentally validated.ConclusionsOur results demonstrated that SphK1 overexpression drives a regulatory network governing miRNA and mRNA expression in PTC cells. We also demonstrated the roles played by miR-144-3p and FN1 in mediating the oncogenic function of SphK1, which enhanced the understanding of the etiology of PTC.