circ_NEK6靶向miR⁃370⁃3p对分化型甲状腺癌^(131)I耐受细胞恶性生物学行为的影响

Effect of circ_NEK6 on malignant biological behavior of ^(131)I⁃resistant differentiated thyroid carcinoma cells via targeting miR⁃370⁃3p

目的探讨circ_NEK6对分化型甲状腺癌(DTC)^(131)I亚热带农业研究B对比上0I耐受细胞恶性生物学行为的影响以及其作用机制。方法采用RT⁃qPCR检测DTC组织和细胞系中circ_NEK6和miR⁃370⁃3p的表达;采用CCK⁃8法、流式细胞术及Transwell法检测^(131)I耐受Res⁃BCPAP细胞增殖、凋亡水平、侵袭和迁移能力;双荧光素酶报告基因验证circ_NEK6与miR⁃370⁃3p的靶向关系。结果circ_NEK6在131I耐受DTC组织中的表达高于^(131)I敏感组织,且其在DTC细胞中的表达高于人甲状腺滤泡上皮正常细胞,尤其是在131I耐受Res⁃BCPAP细胞中的表达水平高于亲本BCPAP细胞。敲降circ_NEK6可抑制Res⁃BCPAP细胞增殖、侵袭和迁移能力,以及诱导细胞凋亡。双荧光素酶报告实验表明,circ_NEK6靶向负调控miR⁃370⁃3p。敲降circ_NEK6靶向上调miR⁃370⁃3p抑制Res⁃BCPAP细胞恶性生物学行为。结论敲降circ_NEK6通过靶向miR⁃370⁃3p增强DTC细胞对^(131)I放射敏感性,提示circ_NEK6可能是131I抵抗DTC患者潜在的生物标志物和治疗靶点。

Objective To investigate the effect of circ_NEK6 on the malignant biological behavior of ^(131)I⁃resistant differentiated thyroid carcinoma(DTC)cells and its possible mechanism.Methods RT⁃qPCR was performed for the detection of circ_NEK6 and miR⁃370⁃3p in DTC tissues and cell lines.The proliferation,apoptosis,migration and invasion abilities of Res⁃BCPAP cells was evaluated by CCK⁃8,flow cytometry,Transwell assay,respectively.Dual⁃luciferase assay was conducted to prove the interaction of circ_NEK6 and miR⁃370⁃3p.Results circ_NEK6 was upregulated in the ^(131)I⁃resistant DTC tissues compared with the 131I⁃sensitive DTC tissues,and its expression in DTC cell lines was higher than that in human thyroid follicular epithelial normal cells,especially in ^(131)I⁃resistant Res⁃BCPAP cells was higher than that in parental BCPAP cells.Knockdown of circ_NEK6 decreased Res⁃BCPAP cell proliferation,migration and invasion,as well as promoted cell apoptosis.Besides,dual⁃luciferase reporter gene assay analysis results showed that circ_NEK6 could directly bind to and negatively regulate miR⁃370⁃3p expression.Functionally,circ_NEK6 suppression inhibited the malignant biological behavior of Res⁃BCPAP cells via targeting miR⁃370⁃3p.Conclusion Inhibition of circ_NEK6 enhanced the radio⁃sensitivity of ^(131)I in DTC cells via targeting miR⁃370⁃3p,indicating that circ_NEK6 may act as a potential biomarker and therapeutic target for DTC patients with 131I resistance.