CAJ | 학술논문

目的探讨RNA干扰(RNAi)抑制信号转导与转录激活因子-3(STAT3)活性对人胰腺癌鸡胚移植瘤周围新生血管的影响及机制.方法构建STAT3短发卡RNA(shRNA)表达载体,稳定转染胰腺癌细胞株SW1990,凝胶电泳迁移率检测实验(EMSA)观察STAT3的DNA结合活性.利用鸡胚绒毛尿囊膜(CAM)建立人胰腺癌鸡胚移植瘤模型,观察移植瘤周围的新生血管生长.逆转录-聚合酶链反应(RT-PCR)检测血管内皮生长因子(VEGF)和基质金属蛋白酶-2(MMP-2)的mRNA表达.结果 RNAi抑制STAT3后,SW1990细胞STAT3活性下降86%;移植瘤体积缩小,为(15.32±2.56)mm3,移植瘤周围新生血管减少;VEGF和MMP-2的mRNA表达分别下降54%和72%.结论 STA3 shRNA表达载体能有效抑制STAT3活性,通过下调VEGF和MMP-2表达,抑制人胰腺癌鸡胚移植瘤周围新生血管.
목적 탐토RNA간우(RNAi)억제신호전도여전록격활인자-3(STAT3)활성대인이선암계배이식류주위신생혈관적영향급궤제.방법 구건STAT3단발잡RNA(shRNA)표체재체,은정전염이선암세포주SW1990,응효전영천이솔검측실험(EMSA)관찰STAT3적DNA결합활성.이용계배융모뇨낭막(CAM)건립인이선암계배이식류모형,관찰이식류주위적신생혈관생장.역전록-취합매련반응(RT-PCR)검측혈관내피생장인자(VEGF)화기질금속단백매-2(MMP-2)적mRNA표체.결과 RNAi억제STAT3후,SW1990세포STAT3활성하강86%;이식류체적축소,위(15.32±2.56)mm3,이식류주위신생혈관감소;VEGF화MMP-2적mRNA표체분별하강54%화72%.결론 STA3 shRNA표체재체능유효억제STAT3활성,통과하조VEGF화MMP-2표체,억제인이선암계배이식류주위신생혈관.
Objective To investigate the effect and mechanism of RNA interference (RNAi)-mediated signal transducers and activators of transcription 3 ( STAT3 ) activity inhibition on the neoangiogenesis of the human pancreatic cancer in chick embryo chorioallantoic membrane (CAM). Methods STAT3 shRNA expression vector was stably transfected to SW1990 cells. STAT3 DNA-binding activity was examined using electrophoretic mobility shift assay (EMSA). The angiogenesis ability of SW1990 cells was determined by CAM model. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the mRNA expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase (MMP)-2. Results STAT3 DNA-binding activity was decreased by 86% after stable transfection of STAT3 shRNA expressing vectors. Inhibition of STAT3 activity with RNAi significantly inhibited the neoangiogenesis ability of SW1990 cells in CAM and decreased the tumor volume to ( 15.32 ±2. 56) mm3. Moreover, the relative VEGF and MMP-2 mRNA expression in SW1990-RNAi cells was reduced by 54% and 72% as compared with that of the parental SW1990 cells, respectively. Conclusion Inhibition of STAT3 activity with RNAi can significantly inhibit the neoangiogenesis ability of pancreatic cancer through down-regulating VEGF and MMP-2.