miRNA-126通过调节notch-1/Akt信号通路调控甲状腺癌细胞SW579增殖、凋亡及迁移

miRNA-126 regulates the proliferation, apoptosis and migration of thyroid cancer cell SW579 by regulating the notch-1/Akt signaling pathway

目的探索miRNA-126对甲状腺癌SW579细胞增殖、凋亡及迁移的影响,并进一步探索其机制。方法体外培养甲状腺癌SW579细胞,利用qPCR检测人甲状腺正常Nthy-ori3-1细胞和SW579细胞miRNA-126的表达;将细胞分为空白对照组、实验组及阴性对照组,分别不做任何处理、空质粒载体进行转染、转染miRNA-126表达质粒;通过转染质粒构建miRNA-126过表达的SW579细胞;利用CCK-8实验检测细胞增殖;Transwell小室实验检测细胞迁移和侵袭;流式细胞术检测细胞凋亡及活性氧水平变化;Western blot检测Notch-1/Akt通路相关蛋白表达,利用SPSS 21.0统计软件通过正态分布和t检验来印证。结果SW579细胞中miRNA-126表达量为(0.25±0.07),与Nthy-ori3-1细胞相比差异具有统计学意义(P<0.001);空白对照组、实验组及阴性对照组细胞存活率为(105.70±7.61)、(98.60±5.42)及(62.70±3.82);细胞凋亡率为(9.14±0.83)、(12.28±1.34)及(36.39±3.21)(P均<0.05);细胞迁移率为(34.51±2.45)、(33.29±3.17)及(11.22±1.23)(P均<0.05);活性氧水平为(1.02±0.07)、(1.08±0.11)及(6.54±0.74)(P均<0.05)。结论过表达miRNA-126能通过上调细胞内活性氧水平抑制Notch-1/Akt通路,抑制SW579细胞的增殖、迁移、诱导SW579细胞发生凋亡,为miRNA-126在甲状腺癌的研究中提供一定的理论基础。

Objective To investigate the effect of miRNA-126 on proliferation,apoptosis and migration of thyroid cancer SW579 cells,and further to explore its mechanism.Methods Thyroid cancer SW579 cells were cultured in vitro,and the expression of miRNA-126 in normal thyroid Nthy-ori3-1 cells and SW579 cells was detected by qPCR.The cells were divided into blank control group(without any treatment),experimental group(transfected with miRNA-126 expression plasmid)and negative control group(transfected with empty plasmid vector).SW579 cells with overexpression of miRNA-126 were constructed by transfection of plasmids.CCK-8 test was used to detect cell proliferation;Transwell cell test was used to detect cell migration and invasion;Flow cytometry was employed to detect apoptosis and changes in reactive oxygen levels;Western blot was used to detect Notch-1/Akt pathway related protein expression.Results The expression level of miRNA-126 in SW579 cells was 0.25±0.07,and the difference was statistically significant compared with that in Nthy-ori3-1 cells(P<0.001).The cell survival rates of blank control group,experimental group and negative control group were(105.70±7.61),(98.60±5.42)and(62.70±3.82);The apoptosis rates were(9.14±0.83),(12.28±1.34)and(36.39±3.21)(all P<0.05);The cell migration rates were(34.51±2.45),(33.29±3.17)and(11.22±1.23)(all P<0.05);The levels of ROS were(1.02±0.07),(1.08±0.11)and(6.54±0.74)(all P<0.05).Conclusion Overexpression of miR-NA-126 can inhibit Notch-1/Akt pathway by up-regulating intracellular reactive oxygen species,inhibit the proliferation and migration of SW579 cells,and induce apoptosis in SW579 cells,which provides a certain basis for the study of miRNA-126 in thyroid cancer.