【摘要】 目的:研究鸟嘌呤核苷酸交换蛋白100(GEP100)基因沉默对胰腺癌细胞体外侵袭能力的影响。方法:将质粒pSuper-retro-puro-GEP100转染入胰腺癌AsPC-1细胞株,通过RNA干扰建立GEP100基因稳定沉默的细胞克隆。以基质侵润实验检测细胞体外侵袭能力,以过河实验检测细胞移动能力,以Western blotting检测细胞上皮型钙黏连蛋白(E-cadherin)的表达。结果:沉默GEP100表达后,细胞体外侵袭能力明显受抑,基质侵润实验显示,GEP100干扰组、空载体对照组、未转染组的穿膜细胞分别为每视野46.62±5.25、115.40±12.46、111.82±10.82,干扰组与空载体对照组、未转染组间均存在明显差异(P<0.01);细胞的移动能力仅受到轻微影响,GEP100干扰组、空载体对照组、未转染组细胞的过河时间分别为(52.68±4.12)h、(47.56±3.42)h、(48.60±5.24)h,各组间无明显差异;沉默GEP100促使细胞从间皮形态向上皮形态转化,并显著增加了E-cadherin蛋白表达(P<0.05)。结论:沉默GEP100可明显抑制胰腺癌细胞的侵袭能力,而对细胞的移动能力无明显影响,E-cad-herin表达上调,细胞表现出间皮向上皮形态转化倾向,提示GEP100可能通过调节E-cadherin的表达来影响细胞间黏附连接的功能,从而在胰腺癌的侵润转移中发挥重要作用。更多还原

【Abstract】 AIM:To study the effects of down-regulation of guanine nucleotide exchange protein 100 (GEP100) on the invasive ability of pancreatic cancer cell AsPC-1 in vitro.METHODS:The clone of AsPC-1 cells with stable knock-down of GEP100 by transfection of pSuper-retro-puro-GEP100 was established.The invasive ability was evaluated by matrigel invasion assay and the migratory ability of the cells was examined by crossing-river test.The protein expression of E-cadherin was determined by Western blotting.RESULTS:The invasive ability was inhibited significantly in matrigel invasion assay (P<0.01).The penetrated cells were 46.62 ± 5.25 /field in GEP100 knock-down group,115.40 ± 12.46 /field in mock group and 111.82 ± 10.82 /field in non-transfected group,respectively.The migratory ability of the cells was just inhibited slightly,showing a crossing-river time period of (52.68 ± 4.12) h,which was a little bit longer than that in non-transfected group (48.60 ± 5.24) h and mock group (47.56 ± 3.42) h without statistical difference.Down-regulation of GEP100 resulted in a morphological change of AsPC-1 cells from mesenchymal type to epithelial type and an obvious up-regulation of E-cadherin protein was observed (P<0.05).CONCLUSION:Suppression of GEP100 inhibits the invasive ability of AsPC-1 cells significantly without obvious influence on the migratory ability.The cells show a transformation from mesenchymal type to epithelial type with increased E-cadherin expression.The above results indicate that GEP100 might play an important role in the invasive ability of pancreatic cells through regulation of the E-cadherin expression.更多还原