ezrin和merlin基因在胰腺癌细胞中的表达及其意义

Expression and significance of ezrin,merlin genes in the pancreatic adenocarcinoma cell

目的:研究ezrin和merlin基因在胰腺癌细胞中的表达情况及其对胰腺癌细胞生物学行为的影响。方法:应用RT-PCR与Western blotting检测ezrin和merlin基因在胰腺导管上皮细胞及胰腺癌细胞中的mRNA和蛋白表达水平;利用脂质体分别将ezrin和merlin基因转染入SW1990细胞株中,应用MTT实验、流式细胞术、迁移实验和细胞黏附实验观察转染基因对细胞增殖、迁移和黏附能力,细胞周期及凋亡的影响。结果:ezrin和merlin基因在8株细胞中均有不同程度的表达,其表达水平与胰腺癌细胞的分化程度无关。与对照组相比,转染ezrin和merlin基因的SW1990细胞增殖速度缓慢(P<0.01);细胞周期表现为G1期细胞增多,S期细胞减少(均P<0.05);细胞与基质的黏附能力明显下降(P<0.01);且转染merlin的SW1990细胞其迁移能力亦有明显降低(P<0.05),而转染ezrin的SW1990细胞其迁移能力无明显统计学意义的改变(P>0.05)。结论:ezrin和merlin蛋白在胰腺癌细胞增殖、进展过程中可能发挥着负性调节作用。

Objective To investigate the expression of ezrin and merlin genes in some pancreatic adenocarcinoma cells and the influence on the biological behaviors of pancreatic adenocarcinoma cell. Methods Expression of ezrin and merlin genes in pancreas ductus endothelial cell HPDE and pancreatic adenocarcinoma cells were detected using RT- PCR and Western blotting analysis. The recombinant plasmids （pcDNA3. 1 ＋ HA-ezrin and pcDNA3. 1 ＋ HA-merlin） were transfected into SW1990 by liposome. We observed the biological behaviors of transfected cells using MMT method, flow cytometry, transwell test and cell matrix adhesion experiment to detect the growth and proliferation, the cell cycle and apoptosis, the migration ability and matrix adhesion ability of the transfected cells. Results There was different degree expression of ezrin and merlin gene in all eight kinds of cells, and there was no relationship between the expression levels of the genes and the differentiation grades of the cell lines. Compared with control groups,SW1990-ezrin and SW1990- merlin showed slower proliferation pattern （ P 〈 0.01 ）. Flow cytometry results revealed that the exogenous ezrin and mer- lin could accumulate cells in G0/G1 stage and decrease cell amounts in S phase （P 〈 0.05 ）. Matrix adhesion ability of the transfected cell significantly decreased（P 〈0.01）. The migration ability of the merlin-transfected cell was remark- ably lower than untransfected cell, but there was no significant difference between untransfected cell and ezrin-transfected cell （P 〉 0.05 ）. Conclusion Ezrin and merlin may play a down-regulation role in the proliferation and progression of pancreatic adenocarcinoma cell.