目的：研究桥接整合因子1（bridging intergrator-1,Bin1）基因过表达对非小细胞肺癌细胞株A549细胞迁移和侵袭能力的影响，并初步探讨其作用机制。方法：通过基因转染技术，利用阳离子脂质体将含有人全长Bin1基因序列的真核表达质粒CMV-MCS-GFP-SV40-Neomycin-Bin1转染到A549细胞株，分别设置空白对照组及空质粒转染组，利用RT-PCR和Western blotting分别检测各处理组细胞中Bin1基因和蛋白表达水平。通过细胞划痕实验、Transwell侵袭实验分别检测Bin1过表达对A549细胞迁移、侵袭能力的影响；Western blotting实验检测Bin1过表达对A549细胞内NF-κB磷酸化水平和迁移相关蛋白E-钙黏着蛋白、N-钙黏着蛋白、MMP-9表达水平的影响。结果：与空白对照组和空质粒转染组相比，Bin1转染组A549细胞中Bin1基因和蛋白表达水平均明显升高（P<0.05）；Bin1转染组细胞迁移、侵袭能力均较空白质粒组和空白对照组明显下降\[穿膜细胞数：(50.50±3.15) vs (124.00±4.25), (130.00±4.37)个；均P<0.05\]；与空白转染组和空白对照组相比，Bin1转染组细胞内NF-κB表达水平明显上调（P<0.05）而p-NF-κB表达明显下调（P<0.05），N-钙黏着蛋白、MPP-9明显下调（P<0.05），E-钙黏着蛋白明显上调（P<0.05）。结论： Bin1过表达可以抑制A549细胞的迁移及侵袭能力，其机制可能与NF-κB途径的失活及细胞迁移侵袭相关蛋白表达变化有关。

Objective:To study the effect of bridging intergrator-1 (Bin1) gene over-expression on migration and invasion abilities of non-small cell lung cancer A549 line cell, and initially discuss mechanism of its action. Methods: Eukaryotic expression plasmid CMV-MCS-GFP-SV40-Neomycin-Bin1 containing full length Bin1 gene sequence was transfected into the A549 cell using cationic liposomes and gene transfection technology (as Bin1 transfection group), as well as blanck control and empty plasmid transfection groups were set up. Expressions of Bin1mRNA and protein in various treatment groups were detected with PT-PCR and Western blotting assays. Effects of Bin1over-expression on migration and invasion abilities of the A549 cell were examed by scarification and Transwell invasion tests respectively. Western blotting assay was used to detect effects of Bin1over-expression on NF-κB phosphorylation level and expressions of migration-related proteins, E-cadherin, N- cadherin and MMP-9, in the A549 cell.Results: Comparing with blanck control and empty plasmid transfection groups, expressions of Bin1 mRNA and protein in the A549 cell of Bin1 transfection group increased significantly (all P<0.05). Cell abilities of migration and invasion in Bin1transfection group were significantly lower than those in empty plasmid transfection and blanck control groups (number of penetrating cells: \[50.50±3.15\] vs \[124.00±425\], \[130.00±4.37\], all P<0.05). In the A549 cell of Bin1transfection group, expressions of NF-κB and E-cadherin obviously increased (all P<0.05), and expressions of p-NF-κB, N- cadherin and MMP-9 significantly decreased (all P<0.05), compared to empty plasmid transfection and blanck control groups. Conclusion: Over-expression of Bin1 could inhibit abilities of migration and invasion of the A549 cell, and its mechanism might be associated with inactivation of NF-κB pathway and expression changes of cell migration and invasion-related proteins.

国家自然科学基金青年科学基金资助项目（No.81201607）；河北省杰出青年基金资助项目（No. H2014206320）；河北省高层次人才培养项目资助（No.A201401040）