白头翁皂苷D通过调控circ_0001178/miR-885-5p对甲状腺癌细胞的生物学行为的影响

Effect of pulsatilla saponin D on the biological behaviors of thyroid cancer cells by regulating the circ_0001178/miR-885- 5p axis

目的探讨白头翁皂苷D对甲状腺癌细胞生物学行为的影响及其对circ_0001178/miR-885-5p分子轴的调控作用。方法不同浓度的白头翁皂苷D处理人甲状腺癌细胞SW579,si-NC、si-circ_0001178、miR-NC、miR-885-5p mimics转染至SW579细胞,pcDNA-circ_0001178、anti-miR-885-5p转染至SW579细胞后加入白头翁皂苷D处理;qRT-PCR检测circ_0001178与miR-885-5p的表达量;MTT、平板克隆形成实验、Transwell小室实验分别检测细胞增殖、克隆形成、迁移及侵袭;双荧光素酶报告基因实验检测circ_0001178与miR-885-5p的靶向关系。结果白头翁皂苷D可降低circ_0001178的表达量、细胞活力及N-cadherin蛋白水平(P<0.05),可抑制克隆形成数、迁移及侵袭,还可促进miR-885-5p表达及E-cadherin的表达(P<0.05),且呈剂量依赖性;转染si-circ_0001178或转染miR-885-5p mimics可抑制细胞增殖、克隆形成、迁移及侵袭;circ_0001178能够靶向调控miR-885-5p;转染pcDNA-circ_0001178或转染anti-miR-885-5p均可逆转白头翁皂苷D对细胞生物学行为的作用。结论白头翁皂苷D可通过调控circ_0001178/miR-885-5p分子轴而抑制甲状腺癌细胞增殖、克隆形成、迁移及侵袭。

Objective To explore the effect of pulsatilla saponin D on the biological behaviors of thyroid cancer cells by regulating the circ_0001178/miR-885-5p axis.Methods Human thyroid cancer cells SW579 were transfected with si-NC,si-circ_0001178,miR-NC,miR-885-5p mimics,pcDNA-circ_0001178,or anti-circ_0001178,followed by the treatment of pulsatilla saponin D at varying concentrations.Relative levels of circ_0001178 and miR-885-5p were detected by quantitative reverse transcription polymerase chain reaction(qRT-PCR).Cell proliferation and colony formation were examined by MTT assay and colony formation assay.Cell migration and invasion were examined by Transwell assay.The targeting relationship between circ_0001178 and miR-885-5p was tested by dual-luciferase reporter assay.Results Treatment of pulsatilla saponin D significantly downregulated circ_0001178 and N-cadherin,reduced cell viability,number of colonies and migratory and invasive cell numbers,and upregulated miR-885-5p and E-cadherin in a dose-dependent manner(P<0.05).Transfection of si-circ_0001178 or miR-885-5p mimics inhibited cell proliferation,colony formation,migration and invasion.Circ_0001178 provided a targeting regulation of miR-885-5p.Transfection of pcDNA-circ_0001178 or anti-circ_0001178 reversed the regulatory effects of pulsatilla saponin D on behaviors of SW579 cells.Conclusion Pulsatilla saponin D inhibits proliferation,colony formation,migration and invasion of thyroid cancer cells by regulating the circ_0001178/miR-885-5p axis.