激活和抑制Stat 3信号途径对胰腺癌细胞侵袭能力的影响

目的:通过激活和阻断人胰腺癌细胞中的Stat3信号转导通路,观察细胞株侵袭能力的变化并探讨其作用机制.方法:采用IL-6处理人胰腺癌细胞Capan-2,AG490处理人胰腺癌细胞SW1990后,MTT法检测细胞的增殖状态,免疫细胞化学法和Western 印迹法检测p-Stat3的表达,实时荧光定量PCR(real-time fluorogentic quantitative PCR, RFQ-PCR)和Western 印迹法检测血管内皮生长因子(vascular endothelial growth factor,VEGF)、基质金属蛋白酶-2(matrix metalloproteinase 2,MMP-2) mRNA及其蛋白的表达,体外侵袭实验检测细胞的侵袭能力.结果:IL-6可促进Capan-2细胞的增殖能力提高(P<0.05),p-Stat3的表达增强,VEGF和MMP-2 mRNA和蛋白表达明显升高(P<0.05),细胞侵袭能力增强.AG490可抑制SW1990细胞株的增殖(P<0.05),p-Stat3的表达下降,VEGF和MMP-2 mRNA和蛋白表达明显下降(P<0.05),侵袭能力减弱.结论:Stat3信号转导通路在胰腺癌侵袭过程中起着重要作用,以Stat3信号转导通路为靶点的基因治疗可能为胰腺癌治疗提供新的方向.

Effects of activation and inhibition of Stat 3 signaling pathway on invasion of human pancreatic cancer cells

Objective: In order to investigate the effects of activating and blocking Stat3 signaling pathway on invasion ability of human pancreatic cancer cells and explore its action mechanism.Methods:Human pancreatic cancer Capan-2 cells were treated with IL-6. SW1990 human pancreatic cancer cells were treated with AG490. Cell proliferation was measured by MTT assay. Western blotting and immunocytochemistry were performed to detect expression of phosphorylated Stat3 (p-Stat3) protein. Real-time fluorogentic quantitative PCR (RFQ-PCR) and Western blotting were used to detect the mRNA and protein expression of VEGF and MMP-2 mRNA, respectively. The invasion abilities of SW1990 and Capan-2 cells were determined by cell invasion assay in vitro. Results:IL-6 stimulated the proliferation of Capan-2 cells (P<0.05), elevated the expression of p-Stat3, increased the mRNA and protein expressions of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 2 (MMP-2) (P<0.05), and enhanced the invasion ability of Capan-2 cells. AG490 inhibited the proliferation of SW1990 cells (P<0.05), down-regulated the expression of p-Stat3, markedly decreased the mRNA and protein expression of VEGF and MMP-2 (P<0.05), and weakened the invasion ability of SW1990 cells. Conclusion:Stat 3 signaling pathway plays an important role in the invasion and metastasis of pancreatic cancer. Stat 3 signaling transduction pathway may provide a novel therapeutic target for the treatment of pancreatic cancer.