Emerging roles of circRNA_NEK6 targeting miR-370-3p in the proliferation and invasion of thyroid cancer via Wnt signaling pathway
Emerging roles of circRNA_NEK6 targeting miR-370-3p in the proliferation and invasion of thyroid cancer via Wnt signaling pathway
复制标题
DOI：
10.1080/15384047.2018.1480888
复制
发表时间：
2018-12-02
影响因子：
3.6
通讯作者：
Deng, Zhiyong
中科院分区：
文献类型：
--
作者：
Chen, Fukun;Feng, Zhiping;Deng, Zhiyong
目的：目的鉴定甲状腺癌中显著改变的circRNAs和mRNAs，研究其靶向miRNAs并确定其生物学功能。研究方法：采用基因芯片分析和基因集富集分析（GSEA）技术，筛选甲状腺癌中差异表达的circRNA、mRNA和通路。通过Pearson相关分析找出相关的circRNA和mRNA。通过Targetscan、米兰达和HMDD分析筛选出与甲状腺癌相关的circNEK 6和FZD 8的共同靶点miRNAs。采用qRT-PCR和western blot检测甲状腺癌组织和细胞中mRNA和蛋白的表达。通过RNase R消化和核酸电泳证实了CircRNA。通过双荧光素酶报告基因测定验证靶关系。MTT法检测细胞活力，Transwell法检测细胞侵袭力，流式细胞仪检测细胞凋亡。结果：CircNEK 6和FZD 8在甲状腺癌中显着上调，且具有强相关性。Wnt信号通路在甲状腺癌中被激活。miR-370- 3 p是circNEK 6和FZD 8的共同靶点，在甲状腺癌中表达下调。过表达circNEK 6和FZD 8可促进甲状腺癌细胞的生长和侵袭，而上调miR-370- 3 p可抑制甲状腺癌的进展并抑制Wnt信号通路。miR-370- 3 p对甲状腺癌细胞的作用可以通过circNEK 6或FZD 8来挽救。结论：CircNEK 6通过上调FZD 8，并以miR-370- 3 p为靶点激活Wnt信号通路，促进甲状腺癌的进展。
Objective: To identify the significantly altered circRNAs and mRNAs in thyroid cancer, investigate their target miRNAs and determine their biological functions. Methods: The differentially expressed circRNAs, mRNAs and pathways in thyroid cancer were identified by microarray analysis and gene set enrichment analysis (GSEA). The correlative circRNAs and mRNAs were found out through Pearson correlative analysis. The common target miRNAs of circNEK6 and FZD8 related to thyroid cancer was screened out through Targetscan, miRanda and HMDD analysis. The mRNA and protein expressions in thyroid cancer tissues and cells were detected by qRT-PCR and western blot. CircRNA was confirmed by the RNase R digestion and nucleic acid electrophoresis. The target relationships were verified by the dual luciferase reporter assay. Cell viability, invasion and apoptosis were determined by MTT assay, Transwell assay and flow cytometry, respectively. Results: CircNEK6 and FZD8 were significantly up-regulated in thyroid cancer, with strong correlations. The Wnt signaling pathway was activated in thyroid cancer. MiR-370-3p was the common target miRNA of circNEK6 and FZD8, and it was down-regulated in thyroid cancer. Overexpression of circNEK6 and FZD8 could promote the growth and invasion of thyroid cancer cells, while up-regulation of miR-370-3p could suppress thyroid cancer progression and inhibit the Wnt signaling pathway. MiR-370-3p's effect on thyroid cancer cells could be rescued by circNEK6 or FZD8. Conclusion: CircNEK6 promoted the progression of thyroid cancer through up-regulating FZD8 and activating Wnt signaling pathway by targeting miR-370-3p.