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

夏迎晨 甄杰 叶飞* 郭惠明 张丽娟

doi:10.16098/j.issn.0529-1356.2018.05.011

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解剖学报 ›› 2018, Vol. 49 ›› Issue (5) : 630-635. DOI: 10.16098/j.issn.0529-1356.2018.05.011

肿瘤生物学

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

夏迎晨¹ 甄杰² 叶飞^2* 郭惠明² 张丽娟²

作者信息 +

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

XIA Ying-chen¹ ZHEN Jie² YE Fei ^2* GUO Hui-ming² ZHANG Li-juan²

Author information +

文章历史 +

摘要

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。

Abstract

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.

导出引用

夏迎晨甄杰叶飞郭惠明张丽娟. 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[J]. 解剖学报. 2018, 49(5): 630-635 https://doi.org/10.16098/j.issn.0529-1356.2018.05.011

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. 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[J]. Acta Anatomica Sinica. 2018, 49(5): 630-635 https://doi.org/10.16098/j.issn.0529-1356.2018.05.011

参考文献

［1］ Arrieta O, Anaya P, Morales-Oyarvide Ⅴ,et al.Cost-effectiveness analysis of EGFR mutation testing in patients with non-small cell lung cancer (NSCLC) with gefitinib or carboplatin-paclitaxel［J］. Eur J Health Econ,2016,17(7):855-863.

［2］ Mirshahidi HR, Hsueh CT. Updates in non-small cell lung cancer-insights from the 2009 45th annual meeting of the American Societyof Clinical Oncology［J］. J Hematol Oncol,2010,3(1):18.

［3］ Monteiro J, Fodde R. Cancer stemness and metastasis: therapeutic consequences and perspectives［J］. Eur J Cancer, 2010,46(7)):1198-1203.

［4］ Fazi F, Fontemaggi G. MicroRNAs and lymph node metastatic disease in lung cancer［J］. Thorac Surg Clin,2012,22(2): 167-175.

［5］ Inamura K, Ishikawa Y. Lung cancer progression and metastasis from the prognostic point of view［J］. Clin Exp Metastasis,2010,27(6):389-397

［6］ Lee RC, Feinbaum RL, Ambros V. The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14［J］.Cell,1993,75(5):843-854.

［7］ Bartels CL, Tsongalis GJ. MicroRNAs: Novel biomarkers for human cancer［J］.Ann Biol Clin,2010,68(3):263-272.

［8］ Liu X, Sempere LF, Ouyang H, et al. MicroRNA-31 functions as an oncogenic microRNA in mouse and human lung cancer cells by repressing specific tumor suppressors［J］. J Clin Invest, 2010, 120(4):1298-1309.

［9］ Puiss égur MP, Mazure NM, Bertero T, et al. miR-210 is overexpressed in late stages of lung cancer and mediates mitochondrial alterations associated with modulation of HIF-1 activity［J］. Cell Death Differ,2011, 18(3):465-478.

［10］Li Y, Li Y, Liu J, et al. Expression levels of microRNA-145 and microRNA-10b are associated with metastasis in non-small cell lung cancer［J］.Cancer Biol Ther,2016,17(3): 272-279.

［11］Chen Z, Zeng HZ, Guo Y, et al. miRNA-145 inhibits non-small cell lung cancer cell proliferation by targeting c-Myc ［J］. J Exp Clin Cancer Res,2010,29(1): 151.

［12］Zhang Y, Lin Q. MicroRNA-145 inhibits migration and invasion by down-regulating FSCN1 in lung cancer［J］. Int J Clin Exp Med,2015,8(6): 8794-8802.

［13］Wang Y, Sun YM. The impact of microRNA143 and microRNA145 on the growth cycle and apoptosis of the QBC939 in bile duct cancer cells ［J］. Journal of Nanjing Medical University (Natural Science Edition), 2010, 30(5): 612-616. (in Chinese)

王勇，孙跃明. microRNA143、microRNA145对胆管癌细胞株QBC939 生长周期及凋亡的影响［J］. 南京医科大学学报(自然科学版),2010, 30(5): 612-616.

［14］Zhang J,Han C,Wu T. Microrna-26A enhances cholangiocarcinoma growth through degradation of GSK-3β and activation of β-catenin［J］. Hepatology,2011, 54(Suppl 1):1356A-1357A.

［15］Yan X, Chen X, Liang H, et al. miR-143 and miR-145 synergistically regulate ERBB3 to suppress cell proliferation and invasion in breast cancer ［J］. Mol Cancer,2014,13(1),220.［16］Gregersen LH, Jacobsen AB, Frankel LB, et al. MicroRNA-145 targets YES and STAT1 in colon cancer cells ［J］. PLoS One,2010, 5 (1):e8836.

［17］Cho WC, Chow AS, Au JS. MiR-145 inhibits cell proliferation of human lung adenocarcinoms by targeting EGFR and NUDT1［J］. RNA Bio,2011, 8 (1):125-131.

［18］Fang F, Chen CX, Ma CL. Expression and clinical significance of miRNA-145 in different cervical tissues of xinjiang uygur and han women ［J］. Chinese Clinical Study, 2016,29(2):173-177. (in Chinese)

方芳，陈彩霞，马彩玲. 新疆维吾尔族和汉族妇女不同宫颈组织中miRNA-145的表达及临床意义［J］. 中国临床研究,2016,29(2):173-177.

［19］Xing XF，Li ZY. The expression and function of miR-143 and miR-145 in gastric cancer ［J］. Journal of Gastroenterology, 2015, 18(1): 50-53.(in Chinese)

邢晓芳，李子禹. miR-143和miR-145在胃癌中的表达及功能研究［J］. 中华胃肠外科杂志,2015, 18(1): 50-53.

［20］Guo YH, Jiang B, Shi J, et al. EGFR-AKT signaling pathway reduced expression of miRNA-145 in lung cancer cells ［J］. Modern Oncology Medicine, 2013, 21(12):2652-2655. (in Chinese)

郭跃辉，姜斌，时娟，等. EGFR-AKT信号通路下调肺癌细胞miRNA-145的表达［J］. 现代肿瘤医学,2013, 21(12):2652-2655.

［21］Guo W, Ren D, Chen X, et al. HEF1 promotes epithelial mesenchymal transition and bone invasion in prostate cancer under the regulation of microRNA-145 ［J］. J Cell Biochem,2013, 114(7):1606-1615.

［22］Zhao YJ,Chun H,Li XL,et la.Effects of crocin on the cell proliferation and extracellular regulated protein kinases pathway in human umbilical vein endothelial cells［J］.Acta Anatomica Sinica,2017,48(3):282-286. (in Chinese)

赵玉娇,春花,李鑫磊,等. 藏红花素对人脐静脉内皮细胞增殖及细胞外调节蛋白激酶信号通路的影响［J］. 解剖学报,2017,48(3):282-286.

［23］Boufraqech M, Zhang L, Jain M, et al. miR-145 suppresses thyroid cancer growth and metastasis and targets AKT3 ［J］. Endocr Relat Cancer, 2014, 21(4): 517-531.

［24］Guo YH, Zhang C, Shi J, et al. Abnormal activation of the EGFR signaling pathway mediates the downregulation of miR-145 through the ERK1/2 in non-small cell lung cancer ［J］. Oncol Rep, 2014, 31(4): 1940-1946.