中华胰腺病杂志
中華胰腺病雜誌
중화이선병잡지
CHINESE JOURNAL OF PANCREATOLOGY
2011年
5期
318-321
,共4页
胰腺肿瘤%Jagged1%血管生成诱导剂%血管内皮生长因子%RNA干扰
胰腺腫瘤%Jagged1%血管生成誘導劑%血管內皮生長因子%RNA榦擾
이선종류%Jagged1%혈관생성유도제%혈관내피생장인자%RNA간우
Pancreatic neoplasms%Jagged1%Angiogenesis inducing agents%Vascular endothelial growth factor%RNA interference
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.
目的 以RNA榦擾技術靶嚮沉默人胰腺癌細胞株Jagged1基因,觀察細胞VEGF、Angiopoietin2、bFGF、MMP9的分泌及細胞遷移能力的變化.方法 應用靶嚮Jagged1的siRNA、對照siRNA(c-siRNA)轉染人胰腺癌細胞株SW1990和PANC1,實時PCR法和蛋白質印跡法檢測細胞Jagged1 mRNA和蛋白的錶達,ELISA法檢測細胞培養上清中VEGF、angiopoietin2、bFGF、MMP9的分泌量,Transwell小室觀察細胞的遷移能力.結果 SW1990和PANC1細胞均高錶達Jagged1基因.15 nmol/L Jagged1 siRNA轉染胰腺癌細胞72 h後,SW1990、PANC1細胞的Jagged1 mRNA錶達被抑製,分彆為c-siRNA組的(59.62 ±2.75)％和(76.96 ±6.16)％,Jagged1蛋白錶達幾乎完全被抑製.SW1990、PANC1細胞的VEGF分泌量均較c-siRNA組顯著減少[(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蛋白分泌無明顯變化.另外,Jagged1 siRNA轉染後,SW1990細胞VEGF mRNA錶達水平為c-siRNA組的(52.26 ±4.85)％ (P＜ 0.05)；PANCI細胞為c-siRNA組的(59.75±4.91)％(P＜0.05).轉染後的SW1990和PANC1細胞的穿膜細胞數分彆為(65.25±5.56)、(57.50±8.58)箇,較c-siRNA組的( 122.25±11.09)、(112.00±12.52)箇顯著減少(P＜0.05).結論 人胰腺癌細胞高錶達Jagged1,沉默Jagged1基因可明顯抑製人胰腺癌細胞VEGF的產生和分泌,併且可降低癌細胞的體外遷移能力.
목적 이RNA간우기술파향침묵인이선암세포주Jagged1기인,관찰세포VEGF、Angiopoietin2、bFGF、MMP9적분비급세포천이능력적변화.방법 응용파향Jagged1적siRNA、대조siRNA(c-siRNA)전염인이선암세포주SW1990화PANC1,실시PCR법화단백질인적법검측세포Jagged1 mRNA화단백적표체,ELISA법검측세포배양상청중VEGF、angiopoietin2、bFGF、MMP9적분비량,Transwell소실관찰세포적천이능력.결과 SW1990화PANC1세포균고표체Jagged1기인.15 nmol/L Jagged1 siRNA전염이선암세포72 h후,SW1990、PANC1세포적Jagged1 mRNA표체피억제,분별위c-siRNA조적(59.62 ±2.75)％화(76.96 ±6.16)％,Jagged1단백표체궤호완전피억제.SW1990、PANC1세포적VEGF분비량균교c-siRNA조현저감소[(867.93±58.69) pg/ml비(1516.24±37.58)pg/ml,(951.13±120.49) pg/ml비(1413.68±33.56) pg/ml,P＜0.05],단angiopoietin2、bFGF、MMP9단백분비무명현변화.령외,Jagged1 siRNA전염후,SW1990세포VEGF mRNA표체수평위c-siRNA조적(52.26 ±4.85)％ (P＜ 0.05)；PANCI세포위c-siRNA조적(59.75±4.91)％(P＜0.05).전염후적SW1990화PANC1세포적천막세포수분별위(65.25±5.56)、(57.50±8.58)개,교c-siRNA조적( 122.25±11.09)、(112.00±12.52)개현저감소(P＜0.05).결론 인이선암세포고표체Jagged1,침묵Jagged1기인가명현억제인이선암세포VEGF적산생화분비,병차가강저암세포적체외천이능력.
Objective 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.
