siRNA下调Ezrin表达对人胰腺癌PANC-1细胞增殖和运动能力的影响

Effect of downregulation of Ezrin expression on human pancreatic cancer PANC-1 cell proliferation and moving capacity

目的:探讨Ezrin蛋白在胰腺癌细胞生长和运动转移过程中的作用。方法:体外培养人胰腺癌PANC-1细胞,针对Ezrin基因设计小干扰RNA(siRNA),并通过脂质体转染入细胞,运用MTT法、流式细胞仪、划痕愈合实验、扫描电镜和Transwell实验方法,观察siRNA下调人胰腺癌PANC-1细胞Ezrin表达后,肿瘤细胞增殖、细胞周期、伪足形成、侵袭和迁移能力的改变。结果:Western印迹分析结果显示,Ezrin siRNA对Ezrin蛋白表达水平有显著的下调作用。下调Ezrin蛋白表达后,PANC-1细胞增殖明显被抑制,G2/M期细胞显著降低;细胞伪足形成显著减少,细胞的运动侵袭能力下降。结论:Ezrin在胰腺癌细胞恶性增殖和运动转移过程中具有重要作用,可能成为抑制胰腺癌侵袭转移的靶点。

Objective：To investigate the effect of siRNA targeted to Ezrin on human pancreatic cancer PANC-1 cell proliferation and metastasis.Methods： Human pancreatic cancer PANC-1 cell was cultured.Four pairs of small interfering RNA（siRNA） targeting Ezrin were designed and transfected into the PANC-1 cells.Western blotting was used to screen the most effective siRNA which had been transfected into the PANC-1 cells.When the expression level of Ezrin was the lowest after successful transfection,cell proliferation was detected by MTT method,cell cycle and apoptosis were detected by flow cytometry,the cell pseudopodia was observed by scanning electron microscopy,and the migration and invasion ability of cells was detected by wound healing assay and transwell assay.Results： Western blotting results revealed that Ezrin siRNA notably regulated down the expression of Ezrin protein.Downregulation of Ezrin expression distinctly inhibited the proliferation of the PANC-1 cells.The cell proportion in G2/M phase,the number of pseudopodia per cell,the migration and invasion ability of PANC-1 cells decreased significantly after siRNA targeting Ezrin successful transfection.Conclusion： Ezrin might play an important role in pancreatic adenocarcinoma cell proliferation and invasion,which indicates that Ezrin may be a novel and effective therapeutic target for preventing pancreatic adenocarcinoma invasion and metastasis.