Bin1基因通过NF-κB途径抑制非小细胞肺癌A549细胞的迁移和侵袭能力

发布时间：2018-04-15 06:35

本文选题：非小细胞肺癌 + A细胞　；参考：《中国肿瘤生物治疗杂志》2016年04期

【摘要】：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(P0.05);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P0.05];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(P0.05)而p-NF-κB表达明显下调(P0.05),N-钙黏着蛋白、MPP-9明显下调(P0.05),E-钙黏着蛋白明显上调(P0.05)。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。
[Abstract]:Aim: to investigate the effect of overexpression of 1(bridging intergrator-1 Bin1 gene on the migration and invasion of non-small cell lung cancer cell line A549, and to explore its mechanism.Methods: the eukaryotic expression plasmid CMV-MCS-GFP-SV40-Neomycin-Bin1 containing human full-length Bin1 gene sequence was transfected into A549 cell line with cationic liposome by gene transfection technique. Blank control group and empty plasmid transfection group were set up respectively.RT-PCR and Western blotting were used to detect the expression of Bin1 gene and protein respectively.The overexpression of Bin1 in A549 cells was detected by cell scratch assay and Transwell invasion assay.The effect of overexpression of Bin1 on the phosphorylation of NF- 魏 B and the expression of E-cadherin and MMP-9 in A549 cells were detected by Western blotting assay.Results: compared with the blank control group and the empty plasmid transfection group, the expression of Bin1 gene and protein in the A549 cells of the BIN1 transfection group were significantly higher than those in the P0.05BIN1 transfection group.The invasiveness was significantly lower than that in the blank plasmid group and the blank control group [the number of perforating cells was 50.50 卤4.25 3.15)vs(124.00 卤4.25, 130.00 卤4.37].Compared with the blank transfection group and the blank control group, the expression level of NF- 魏 B increased significantly (P0.05), and the expression of p-NF- 魏 B down-regulated the expression of P0.05N- calcitonin (MPP-9) down-regulated the expression of P0.05- E-cadherin.Conclusion the overexpression of BIN1 can inhibit the migration and invasion ability of A549 cells, and its mechanism may be related to the inactivation of NF- 魏 B pathway and the changes of cell migration and invasion related protein expression.
【作者单位】：河北医科大学第四医院生物治疗科;
【基金】：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)~~
【分类号】：R734.2

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