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。
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 sig- nificantly 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 regu- lation of the E - cadherin expression.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2010年第7期1348-1351,共4页
Chinese Journal of Pathophysiology
基金
浙江省自然科学基金资助项目(No.Y2080372)
浙江省科技厅资助项目(No.2009C33151)
