摘要目的 以RNA干扰技术靶向沉默人胰腺癌细胞株Jagged1基因,观察细胞VEGF、Angiopoietin2、bFGF、MMP9的分泌及细胞迁移能力的变化.方法 应用靶向Jagged1的siRNA、对照siRNA(c-siRNA)转染人胰腺癌细胞株SW1990和PANC1,实时PCR法和蛋白质印迹法检测细胞Jagged1 mRNA和蛋白的表达,ELISA法检测细胞培养上清中VEGF、angiopoietin2、bFGF、MMP9的分泌量,Transwell小室观察细胞的迁移能力.结果 SW1990和PANC1细胞均高表达Jagged1基因.15 nmol/L Jagged1 siRNA转染胰腺癌细胞72 h后,SW1990、PANC1细胞的Jagged1 mRNA表达被抑制,分别为c-siRNA组的(59.62 ±2.75)％和(76.96 ±6.16)％,Jagged1蛋白表达几乎完全被抑制.SW1990、PANC1细胞的VEGF分泌量均较c-siRNA组显著减少[(867.93±58.69) pg/ml比(1516.24±37.58)pg/ml,(951.13±120.49) pg/ml比(1413.68±33.56) pg/ml,P＜0.05],但angiopoietin2、bFGF、MMP9蛋白分泌无明显变化.另外,Jagged1 siRNA转染后,SW1990细胞VEGF mRNA表达水平为c-siRNA组的(52.26 ±4.85)％ (P＜ 0.05)；PANCI细胞为c-siRNA组的(59.75±4.91)％(P＜0.05).转染后的SW1990和PANC1细胞的穿膜细胞数分别为(65.25±5.56)、(57.50±8.58)个,较c-siRNA组的( 122.25±11.09)、(112.00±12.52)个显著减少(P＜0.05).结论 人胰腺癌细胞高表达Jagged1,沉默Jagged1基因可明显抑制人胰腺癌细胞VEGF的产生和分泌,并且可降低癌细胞的体外迁移能力.

abstractsObjective To investigate the effects of Jagged1 gene silencing on the expression of VEGF,angiopoietin 2,bFGF,MMP 9,and the migration of human pancreatic cancer cells SW1990 and PANC1.Methods Jagged1 siRNA and control siRNA were transfected into human pancreatic cancer cells SW1990 and PANC1 respectively.The expressions of Jaggedl mRNA and protein were detected by real-time PCR and Western blotting.ELISA was used to detect the concentration of VEGF,angiopoietin 2,bFGF,MMP9 in cell supernatants.Transwell was used to detect the migration of SW1990 and PANC1 cells.Results Jagged1 mRNA and protein were highly expressed in human pancreatic cancer cells SW1990 and PANC1.After transfected with 15 nmol/L Jagged1 siRNA for 72 h,the expression of Jagged1 mRNA in SW1990 was inhibited and reduced by (59.62 ±2.75)％ and in PANC1 reduced by (76.96 ±6.16)％ when compared with that in control siRNA group.The Jagged1 protein expression was almost inhibited.The concentration of VEGF in the culture supernatants of SW1990 and PANC1 cells was significantly decreased[( 867.93 ± 58.69 ) pg/ml vs.(1516.24 ±37.58)pg/ml,951.13 ± 120.49)pg/ml vs.(1413.68 ±33.56)pg/ml,P ＜0.05],but the protein concentration of angiopoietin 2,bFGF,MMP9 were not significantly changed.In addition,after transfected with Jagged1 siRNA,tne expression of VEGF mRNA was (52.26 ± 4.85 )％ of the control levels in SW1990 ( P ＜ 0.05),and (59.75 ± 4.91 ) ％ of the control levels( P ＜ 0.05 ) in PANC1.The number of cell migration of transfected SW1990 and PANC1 was 65.25 ± 5.56 and 57.50 ± 8.58,which were significantly lower than thosein the control siRNA group (122.25±11.09,112.00±12.52,P＜0.05).Conclusions Jagged1 is highly expressed in human pancreatic cancer cells.Jagged1 gene silencing significantly inhibits the production and secretion of VEGF in SW1990 and PANC1 cells,and reduces the migration of cancer cells in vitro.