摘要
目的探讨微RNA(miR)-223-3p对甲状腺癌细胞增殖和凋亡的影响以及潜在的作用机制。方法运用实时定量聚合酶链式反应(qRT-PCR)检测甲状腺癌SW579细胞和正常甲状腺细胞Nthy-ori 3-1中miR-223-3p和MTSS1的表达水平;将miR-NC组(转染miR-NC)、miR-223-3p组(转染miR-223-3 pmimics)、pcDNA组(转染pcDNA)、pcDNA-MTSS1组(转染pcDNA-MTSS1)、anti-miR-NC组(转染anti-miR-NC)、anti-miR-223-3p组(转染anti-miR-223-3p)、miR-NC+WT-MTSS1组(共转染miR-NC和WT-MTSS1)、miR-NC+MUT-MTSS1组(共转染miR-NC和MUT-MTSS1)、miR-223-3p+WT-MTSS1组(共转染miR-223-3p和WT-MTSS1)、miR-223-3p+MUT-MTSS1组(共转染miR-223-3p和MUT-MTSS1)、anti-miR-223-3p+si-NC组(共转染anti-miR-223-3p和si-NC)、anti-miR-223-3p+si-MTSS1组(共转染anti-miR-223-3p和si-MTSS1),均用脂质体法转染至SW579细胞;采用CCK-8法检测各组细胞增殖;流式细胞术检测各组细胞的凋亡;免疫印迹检测MTSS1蛋白表达;双荧光素酶报告基因检测实验检测细胞的荧光活性。结果甲状腺癌SW579细胞于正常甲状腺细胞Nthy-ori 3-1,miR-223-3p的表达水平显著升高,MTSS1 mRNA和蛋白的表达水平显著降低(P<0.05);miR-223-3p可靶向调控MTSS1的表达;抑制miR-223-3p表达、MTSS1过表达均可抑制SW579细胞增殖,促进凋亡。抑制MTSS1表达逆转了抑制miR-223-3p表达对甲状腺癌SW579细胞增殖抑制和凋亡促进的作用。结论miR-223-3p可抑制甲状腺癌SW579细胞的增殖、促进其凋亡,其机制可能与靶向MTSS1有关,将可为甲状腺癌的预防和治疗提供新靶点。
Objective To investigate the effects of microRNA (miR)-223-3p on proliferation and invasion of thyroid cancer cells and the potential mechanism. Methods Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-223-3p and MTSS1 in thyroid cancer SW579 cells and normal thyroid Nthy-ori 3-1 cell-line. The SW579 cells were subject to different protocol of liposome transfection and divided into miR-NC group (transfected with miR-NC), miR-223-3p group (transfected with miR-223-3 mimics), pcDNA group (transfected with pcDNA), pcDNA-MTSS1 group (transfected with pcDNA-MTSS1), anti-miR-NC group (transfected with anti-miR-NC), anti-miR-223-3p group (transfected with anti-miR-223-3p), miR-NC + WT-MTSS1 group (co-transfected with miR-NC and WT-MTSS1), miR-NC+MUT-MTSS1 group (co-transfected with miR-NC and MUT-MTSS1), miR-223-3p+ WT-MTSS1 group (co-transfection with miR-223-3p and WT-MTSS1), miR-223-3p+MUT-MTSS1 (co-transfected with miR-223-3p and MUT-MTSS1), anti-miR-223-3p + si-NC group (co-transfected with anti-miR-223-3p and si-NC) and anti-miR-223-3p+si-MTSS1 (co-transfected with anti-miR-223-3p and si-MTSS1). For each group, cell proliferation was detected by CCK-8 assay, cell aptopsis by flow cytometry, MTSS1 protein expression by Western blotting, and fluorescence cell viability by dual luciferase reporter gene assay. Results Compared with normal thyroid Nthy-ori 3-1 cells, the thyroid cancer SW579 cells showed significantly increased expression level of miR-223-3p and decreased expression levels of MTSS1 mRNA and protein (P<0.05). miR-223-3p may negatively target-regulate the MTSS1 expression. Either suppressed expression of miR-223-3p or overexpression of MTSS1 may inhibit the proliferation and promote apoptosis of SW579 cells. Suppressed MTSS1 expression may regress the inhibitory effect of miR-223-3p down-regulation on proliferation and apoptosis of thyroid cancer SW578 cells. Conclusion miR-223-3p may inhibit the proliferation and promote apoptosis of thyroid cancer SW578 cells. The mechanism may be related to its targeted regulation of MTSS1, which reveals a new target for prevention and treatment of thyroid cancers.
作者
石佩
申虎威
刘海花
Shi Pei;Shen Huwei;Liu Haihua(Department of Endocrinology, Heping Hospital Affiliated to Changzhi Medical College, Changzhi, Shanxi 046000, China)
出处
《中华生物医学工程杂志》
CAS
2019年第3期293-299,共7页
Chinese Journal of Biomedical Engineering
基金
山西省卫生计生委科研课题项目(201602034).
