miR-145 Attenuates Migration and Invasion of Non-small Cell Lung Cancer A549 Cells by Targeting AP4
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摘要:目的
探讨miR-145对非小细胞肺癌迁移与侵袭能力的影响及其可能的作用机制。
方法qRT-PCR法检测非小细胞肺癌组织中miR-145和激活增强子结合蛋白4（activating enhancer binding protein 4, AP4）mRNA的表达。A549细胞转染miR-145 mimic或者AP4 siRNA后，划痕实验与Transwell小室法分别检测A549细胞的迁移与侵袭能力，qRT-PCR法与免疫印迹法（Western blot）检测A549细胞中AP4 mRNA与蛋白表达水平；双荧光素酶报告基因法检测AP4 mRNA与miR-145的关系。
结果与癌旁正常组织相比，非小细胞肺癌组织中miR-145表达明显降低（P < 0.05），而AP4 mRNA和蛋白表达均明显升高（均P < 0.05）；与对照组相比，转染miR-145 mimic后，A549细胞中miR-145表达明显升高（P < 0.05），AP4 mRNA和蛋白水平明显降低（P < 0.05）；A549细胞的迁移数与侵袭能力均显著下降（均P < 0.05）；双荧光素酶报告基因法验证AP4 mRNA是miR-145的靶基因。A549细胞转染AP4 siRNA后，A549细胞的迁移数与侵袭数显著下降（均P < 0.05）。
结论上调miR-145能抑制A549细胞的迁移与侵袭能力，可能与其抑制靶基因AP4表达有关。
Abstract:ObjectiveTo investigate the effect of miR-145 on the migration and invasion of non-small cell lung cancer (NSCLC) cells and possible mechanism.
MethodsThe expression of miR-145 and activating enhancer binding protein 4 (AP4) mRNA in NSCLC tissues were detected by qRT-PCR. After the transfection with miR-145 mimic or AP4 siRNA, the migration and invasion of A549 cells were examined by wound healing and Transwell assay, respectively; The mRNA and protein expressions of AP4 were detected by qRT-PCR and Western blot, respectively. The relationship between AP4 mRNA and miR-145 was detected by double luciferase reporter gene method.
ResultsCompared with adjacent normal tissues, the level of miR-145 was significantly decreased in NSCLC tissues (P < 0.05), however, the mRNA and protein levels of AP4 were significantly increased (P < 0.05). Compared with control group, the level of miR-145 was significantly increased (P < 0.05), the mRNA and protein levels of AP4 were significantly decreased (P < 0.05), and cells migration and invasion were significantly decreased (P < 0.05) in A549 cells after the transfection with miR-145 mimic. Moreover, AP4 was a direct target of miR-145. After the transfection with AP4 siRNA, the migration and invasion of A549 cells were significantly decreased (P < 0.05).
ConclusionmiR-145 upregulation could efficiently attenuate the migration and invasion of A549 cells via inhibiting target gene AP4 expression.
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Key words:
- Lung cancer /
- miR-145 /
- Migration /
- Invasion /
- AP4
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作者贡献赵天增：设计课题、实施实验与文章撰写 杨金华：实施实验 刘向前：参与临床样本收集和进行数据统计 徐萌博：进行数据统计
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图 1 miR-145在非小细胞肺癌组织中的表达及其与AP4表达的相关性(n=40)
Figure 1 miR-145 was widely down-regulated in NSCLC tissues and negatively correlated with AP4 expression (n=40)
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图 2 miR-145过表达抑制A549细胞侵袭与迁移(n=3)
Figure 2 miR-145 overexpression inhibited migration and invasion of A549 cells (n=3)
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