凋亡抑制蛋白c-FLIP(L)对肺癌细胞增殖迁移的影响-东南大学学报医学版

凋亡抑制蛋白c-FLIP(L)对肺癌细胞增殖迁移的影响

单位：南京大学生命科学院医药生物技术国家重点实验室, 江苏南京 210093

分类号：R734.2;R318

出版年·卷·期（页码）：2015·34·第四期（536-540）

摘要：

目的:近来凋亡抑制蛋白c-FLIP(L)作为新的抗肿瘤治疗靶点受到关注,为了探讨c-FLIP(L)在肺癌细胞迁移中的作用,我们建立了c-FLIP(L)高表达稳定细胞株,以比较c-FLIP(L)在肺癌细胞株中表达水平与其细胞迁移能力的关系。方法:采用非小细胞肺癌A549细胞株进行转染c-FLIP(L)真核表达质粒,通过G418筛选出多株c-FLIP(L)高表达单克隆,以此为平台研究c-FLIP(L)在肿瘤细胞迁移过程中的作用。通过实时荧光定量QPCR及Western Blot检测稳定株中c-FLIP(L)表达水平;划痕实验及Transwell法检测c-FLIP(L)高表达对肺癌细胞迁移的影响,并通过荧光骨架蛋白染色观察细胞形态的改变。结果:QPCR和Western Blot显示A549稳定细胞株中c-FLIP(L)均为高表达;Wound healing和Transwell实验表明c-FLIP(L)高表达肺癌细胞迁移能力明显上升,并初步探索了c-FLIP(L)表达对细胞骨架的影响。结论:凋亡抑制蛋白c-FLIP(L)的高表达能促进肺癌A549细胞的迁移。

Objective: Cellular FLICE-like inhibitory protein (c-FLIP(L)) is an important target for cancer therapy. In this study, we established stable cell line in A549 overexpressing c-FLIP(L) and to examine the potential role of c-FLIP(L) in migration of lung cancer cells. Methods: A549 cells was transfected with recombinant plasmid pcDNA3.1-FLIP and then selected with G418. Expressions of c-FLIP(L) in stable cells were detected by QPCR and Western Blot. The effect of c-FLIP(L) on cell migration was determined using Would healing and Transwell assays, respectively. Results: c-FLIP(L) was successfully highly expressed in A549 stable cells, and increased expression of c-FLIP(L) significantly promoted cell migration testified by Wound healing and Transwell assay. Conclusion: c-FLIP(L) is upregualted in A549 cells relative to the migration ability. This finding is helpful for exploring the molecular mechanism of metastasis in lung cancer cells.

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