DLL4-RNAi影响胃癌细胞增殖和迁移

Effect of DLL4-RNAi on proliferation and migration of gastric cancer cells

目的探讨Delta-like配体4(Delta-like ligand 4,DLL4)在胃癌中的表达情况及对胃癌细胞增殖和迁移能力的影响。方法 RT-PCR验证胃癌细胞中DLL4 mRNA的表达。构建DLL4-RNAi质粒转染胃癌AGS细胞,0.2μg/m L嘌呤霉素筛选建立稳定细胞株。MTT检测细胞株细胞增殖情况;划痕实验观察细胞株细胞的迁移情况。结果RT-PCR结果显示,DLL4和血管内皮生长因子(vascular endothelial growth factor,VEGF)mRNA在胃癌AGS细胞中的表达高于正常细胞GES-1中的表达(均P<0.05),且DLL4与VEGF mRNA的表达呈正相关(r=0.617,P=0.032)。DLL4-RNAi(34251)和DLL4-RNAi(34253)质粒转染可抑制胃癌AGS细胞DLL4 mRNA的表达(均P<0.05)。DLL4-RNAi(34251)可抑制VEGF mRNA的表达(P<0.05)。MTT实验结果显示,转染DLL4-RNAi(34253)和DLL4-RNAi(34251)的AGS细胞增殖速率减慢(均P<0.01)。划痕实验结果显示,转染DLL4-RNAi(34253)的AGS细胞的迁移率降低(P<0.01)。结论胃癌AGS细胞中DLL4表达上调,并影响细胞的增殖和迁移。

Objective To investigate the effect of Delta-like ligand 4（ DLL4） on the proliferation and migration of gastric cancer cells. Methods RT-PCR was employed to detect the expression of DLL4 mRNA in gastric cancer cells. The DLL4-RNAi plasmid was constructed and transfected in gastric cancer AGS cells; the transfected AGS cells were screened by 0. 2 μ g/m L purinomycin to establish a stable cell line. MTT assay was used to detect the cell proliferation. Scratch assay was used to observe the cell migration. Results RT-PCR showed that the expression of DLL4 and vascular endothelial growth factor（ VEGF） mRNA in the gastric cancer AGS cells was higher than those in the normal gastric GES-1 cells（ both P〈0. 05）. Moreover,the expression of DLL4 mRNA was positively correlated with VEGF mRNA level（ r = 0. 617,P =0. 032）. RT-PCR showed that DLL4-RNAi（ 34251） and DLL4-RNAi（ 34253） transfection effectively inhibited the expression of DLL4 mRNA in AGS cells（ both P〈0. 05）,and DLL4-RNAi（ 34251） transfection significantly inhibited the expression of VEGF mRNA（ P〈0. 05）. The MTT assay showed that after DLL4-RNAi（ 34253） and DLL4-RNAi（ 34251） transfection,the proliferation rate of AGS cells was significantly reduced（ both P〈0. 01）. The scratch assay showed that the migration rate was also reduced in DLL4-RNAi（ 34253） transfected AGS cells（ P〈0. 01）. Conclusion DLL4 expression is upregulated in gastric cancer AGS cells,which is related to cell proliferation and cell migration ability.The results indicate that DLL4 might be used as a novel target for treatment of gastric cancer.