miR-182表达异常对甲状腺癌细胞株SW579增殖迁移的影响

Effects of miRNA- 182 on the proliferation and migration in thyroid cancer cell line SW579

目的:研究不同病理状态甲状腺组织中miRNA-182(miR-182)的mRNA表达水平,探讨沉默miR-182后,甲状腺癌细胞系SW579的增殖、迁移能力及VEGF和p53的蛋白表达水平变化。方法:实时荧光定量PCR检测miR-182的mRNA表达水平。转染miR-182-inhibitor后,MTT法、流式细胞术和划痕实验分别检测SW579细胞的增殖和迁移能力的变化。免疫印迹法(Western blotting)检测细胞中VEGF和p53的蛋白表达水平。结果:miR-182在正常甲状腺组织和癌旁甲状腺组织中几乎不表达,在甲状腺腺瘤组织和甲状腺癌组织中呈高表达。转染miR-182-inhibitor后,SW579细胞的增殖能力和细胞迁移数显著降低(P<0.05),VEGF和p53的蛋白表达水平显著降低(P<0.05)。结论:miR-182高表达可能与甲状腺癌的发生发展具有潜在联系。下调miR-182可抑制SW579细胞的增殖迁移,其作用机制与降低细胞中VEGF和p53的蛋白表达水平有关。

Objective: To investigate the expression of miRNA- 182( miR- 182) in thyroid tissue under different physiological pathology conditions,and explore the effects of miR- 182 deficiency on proliferation,migration and protein expression of VEGF and p53 in SW579 cells. Methods: The expression of miR- 182 in different thyroid tissues was determined with Real- time quantitative PCR( RT- q PCR). After transfection with miR- 182 inhibitor using lipofectamine in SW579 cells,cell proliferation,migration,and VEGF and p53 expression were examined with MTT assay,wound healing assay,transwell assay and western blotting,respectively. Results: miR- 182 was prominently expressed in thyroid cancer tissues,as compared with normal thyroid tissues( P < 0. 05). Knockdown of miR- 182 inhibited SW579 cell proliferation and migration( P < 0. 05). The protein expression of VEGF and p53 was significantly decreased after transfection with miR- 182 inhibitor. Conclusion: The high expression of miR- 182 may be correlated with the development of thyroid cancer. Decreased VEGF and p53 protein expression may underlie the inhibition of thyroid cancer cell proliferation and migration after deficiency of miR- 182. The results indicated miR- 182 played an important role in thyroid cancer.