MicroRNA-145通过丝裂原活化蛋白激酶和磷脂酰肌醇3激酶/蛋白质丝氨酸苏氨酸激酶通路调控肺癌A549细胞系转移及侵袭

目的 探讨microRNA-145(miR-145)对非小细胞肺癌A549细胞转移、侵袭及对丝裂原活化蛋白激酶(MAPK)和磷脂酰肌醇3激酶/蛋白质丝氨酸苏氨酸激酶（PI3K/AKT）通路的作用。 方法 将非小细胞肺癌A549细胞分成miR-145模拟物（mimics）组和negative-mimics组（miR-NC）以及antago miR-145组（抑制剂组）和antago miR control 组（antago-NC），采用Transwell迁移实验及基质胶侵袭实验等检测miR-145对人非小细胞肺癌A549迁移、侵袭能力的影响；Western blotting方法分析miR-145对MAPK和PI3K/AKT通路的影响。此外，采用细胞外调节蛋白激酶（ERK）及AKT的通路抑制剂分别作用于A549细胞系，检测A549细胞迁移、侵袭能力的改变。 结果 miR-145 mimics组穿过细胞数（90.67±10.33）明显少于miR-NC组（175.33±23.67），miR-145 mimics组穿过基质胶的细胞数（153.33±22.33）少于miR-NC组 (77.33±13.67），P<0.05；antago-NC组通过小室的细胞数量以及穿过基质胶的细胞数量明显少于antago miR-145组（P<0.05），结果说明，miR-145具有抑制非小细胞肺癌A549细胞迁移、侵袭的能力；miR-145 mimics转染可分别抑制A549细胞中90%、78%以及73%的ERK1/2、AKT的ser-473位点和thr-308位点的磷酸化，antago miR-145转染可促进A549细胞中ERK1/2、AKT的ser-473位点和thr-308位点的磷酸化，增加115%、125%以及129%，而当抑制MAPK通路及PI3K/AKT通路的激活后，A549细胞的转移及侵袭能力下降。 结论 miR-145通过MAPK和PI3K/AKT通路调控肺癌A549细胞转移及侵袭。

Effects of microRNA-145 on the migration and invasion of lung cancer cell line A549 through mitogen-activated protein kinase and phosphatidylinositol 3 kinase/protein-serine-threonine kinase pathways

Objective To study the effect of microRNA-145(miR-145) on the migration and invasion and on the mitogen-activated protein kinase(MAPK) and phosphatiolylinositol 3 kinase/protein serine threonine kinase(PI3K/AKT) pathways in non-small cell lung cancer A549 cells. Methods The A549 cells were divided into miR-145 mimics, negative mimics (miR-NC), antago miR-145 and antago-miR control (antago-NC) groups. Transwell migration experiment and matrix matrigel invasion experiment were used to detect the influence of miR-145 on the migration and invasion ability of human non-small cell lung cancer A549. Western blotting method analyzed the effect of miR-145 on MAPK and PI3K/AKT pathways. In addition, the inhibitors of extracellular regulated protein kinase(ERK) and AKT pathways were used to detect the changes of the cell migration and invasion in A549 cell lines. Results The amount of miR-145 mimics group through transwell chambers was significantly less than the miR-NC group (175.33±23.67). The miR-145 mimics group had a lower number of cells through matrigel matrix than miR-NC group (153.33±22.33), P<0.05. When the expression of miR-145 was inhibited, the number of cells in antago-NC group through Transwell chambers or matrigel matrix were significantly less than the antago miR-145 group (245.00±23.00) and (185.00±12.00), P<0.05. The above results proved that miR-145 inhibited cell migration and invasion ability of A549 cells. The transfection of miR-145 mimics respectively inhibited the phosphorylation of ERK1/2, AKT (ser-473) and AKT (thr-308) about 90%, 78% and 73%, while the transfection of antago miR-145 promoted the phosphorylation of ERK1/2, AKT (ser-473) and AKT (thr-308) and rose up to 115%, 125%and 129%. The migration and invasion ability of A549 cells decreased when the activation of MAPK pathway and PI3K/AKT pathways were inhibited. Conclusion miR-145 regulates lung cancer A549 cell migration and invasion through MAPK and PI3K/AKT pathways, which may provide reliable research results for the treatment of lung cancer.