Objective To investigate the relationship between the migration and invasion ability of human pancreatic cancer cells transfected with Notch1 and cytoskeleton proteins.Methods The lentiviral vector carrying Notch1 was transfected into human pancreatic cancer SW-1900 cell line and cultured and passaged by flow cytometry.The cells were divided into blank group (untreated),unloaded group (empty vector transfection) and virus group (lentivirus transfection).The cell migration and cell invasion ability of the three groups were detected by cell scratch and Transwell chamber respectively;The expression levels of Notch1,Fascin and F-actin in different cell lines were detected by Western Blot.Results Pancreatic cancer SW-1900 cell line stably transfected with Notch1 was successfully established.Scratch tests and Transwell cell displayed in blank group,empty load group,virus group were difference.After 24 h,the virus group and the control group had no difference in the distance of scratches spacing,2 groups of SW-1900 cells adhesion to matrigel membrane were few,roughly the same;But 48 h,72 h later in virus group the scratch space was more narrow than in the control group,cells invasion through a large number of films,the difference was statistically significant,and upregulation of Notch1 could obviously promote the migration and the invasion ability of SW-1900 cells (P ＜0.05).With the increase of time,the rate of cell penetrating and the distance between scratches decreased obviously.According to Western Blot results,the expression of the blank group and the group of Notch1,Fascin load and F-actin protein were similar,the difference was not statistically significant (P ＞0.05),but compared with the other 2 groups,the expression of Notch1,Fascin,F-actin protein of the virus group significantly increased (P ＜0.05).Conclusion High expression of Notch1 protein in human pancreatic cancer cells can enhance cell migration,invasion ability and the content of Fascin and F-actin proteins in cells.One of the mechanisms underlying the enhancement of invasion and metastasis of pancreatic cancer cells may be the enhancement of motility of microfilaments and microtubules in cells.%目的 探讨人胰腺癌细胞转染Notch1后细胞迁移及侵袭能力变化与骨架蛋白的相关性.方法 将携带Notch1的慢病毒载体转染人胰腺癌SW-1900细胞株,经流式细胞筛选后培养、传代.将细胞分为空白组(未经处理)、空载组(空载体转染)和病毒组(慢病毒转染),3组细胞分别利用细胞划痕和Transwell小室检测细胞迁移、细胞侵袭能力;采用Western Blot检测不同组细胞株中Notch1、Fascin、F-actin蛋白表达水平变化.结果 成功建立稳定转染Notch1的胰腺癌SW-1900细胞株.空白组、空载组、病毒组划痕试验及Transwell小室显示:24 h后病毒组和空白组划痕间距无差异,2组细胞粘附到基质胶基底膜的数目均很少,大体相同;而48,72 h后病毒组较空白组划痕间距变窄,细胞侵袭透膜数较多,差异有统计学意义,上调Notch1后能明显促进SW-1900细胞的迁移和侵袭能力(P＜0.05).随着时间延长,细胞穿膜率增高和划痕间距明显减小;Western Blot检测结果显示,空白组和空载组中Notch1、骨架蛋白Fascin、F-actin的蛋白表达相近,差异无统计学意义(P＞0.05),但病毒组与其他2组相比,Notch1、Fascin、F-actin的蛋白表达明显增高,差异有统计学意义(P＜0.05).结论 人胰腺癌细胞中Notch1蛋白高表达可以增强细胞迁移、侵袭能力和细胞中Fascin和F-actin蛋白含量.胰腺癌细胞侵袭和转移能力的增强其机制之一可能是通过加强细胞中微丝微管运动性来实现的.
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