CAJ | 학술논문

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。
목적：근래조망억제단백c－FLIP（ L）작위신적항종류치료파점수도관주，위료탐토c－FLIP（ L）재폐암세포천이중적작용，아문건립료c－FLIP（ L）고표체은정세포주，이비교c－FLIP（ L）재폐암세포주중표체수평여기세포천이능력적관계。방법：채용비소세포폐암A549세포주진행전염c－FLIP（ L）진핵표체질립，통과G418사선출다주c－FLIP（ L）고표체단극륭，이차위평태연구c－FLIP（ L）재종류세포천이과정중적작용。통과실시형광정량QPCR급Western Blot검측은정주중c－FLIP（ L）표체수평；화흔실험급Transwell법검측c－FLIP（ L）고표체대폐암세포천이적영향，병통과형광골가단백염색관찰세포형태적개변。결과：QPCR화Western Blot현시A549은정세포주중c－FLIP（ L）균위고표체；Wound healing화Transwell실험표명c－FLIP （ L）고표체폐암세포천이능력명현상승，병초보탐색료c－FLIP（ L）표체대세포골가적영향。결론：조망억제단백c－FLIP（ L）적고표체능촉진폐암A549세포적천이。
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 <br> finding is helpful for exploring the molecular mechanism of metastasis in lung cancer cells.