摘要
目的研究Janus激酶抑制剂AG490对人肝癌SMMC-7721细胞侵袭转移能力的影响,探讨JAK2/STAT3信号通路在肝癌侵袭调控中的作用。方法实验分为对照组、实验组(5、10μmol/L AG490处理的SMMC-7721细胞)。采用Transwell小室检测肝癌细胞的侵袭能力,RT-PCR检测MMP-2 mRNA和VEGF mRNA表达的变化,Western blot检测STAT3、MMP-2、VEGF蛋白的表达和活化。结果 AG490处理人肝癌细胞48 h后,对照组透膜细胞数为(74.6±6.3)/视野,5μmol/L AG490组透膜细胞数为(58.2±7.2)/视野,10μmol/L组透膜细胞数为(38.2±5.7)/视野,实验组与对照组相比,细胞侵袭人工基底膜的能力显著降低(P<0.05);与对照组相比,RT-PCR显示实验组MMP-2 mRNA和VEGF mRNA表达降低(P<0.05),Western blot显示实验组p-STAT3的活化降低,MMP-2及VEGF蛋白表达降低(P<0.05)。结论阻断STAT3蛋白的激活,能够降低MMP-2及VEGF基因蛋白的表达,抑制肝癌细胞在体外的侵袭能力。
Objective To study the effect of Janus kinase（JAK） inhibitor AG490 on the invasion and metastasis of human liver cancer SMMC-7721 cells and the role of JAK-STAT3 signaling pathway in regulation of liver cancer invasion.Methods SMMC-7721 cells were divided into control group,5 μmol/L AG490 treatment group and 10 μmol/L AG490 treatment group.Invasion and metastasis of liver cancer SMMC-7721 cells were detected with transwell chamber.Expression of MMP-2 mRNA and VEGF mRNA in liver cancer SMMC-7721 cells was detected by RT-PCR.Expression and activation of STAT3,MMP-2 and VEGF proteins in liver cancer SMMC-7721 cells was detected by Western blotting.Results The number of membrane-permeating SMMC-7721 cells was 74.6±6.3,58.2±7.2,and 38.2±5.7 in control group,5 μmol/L AG490 treatment group,and 10 μmol/L AG490 treatment group,respectively,48 h after AG490 treatment.The ability of SMMC-7721 cells to invade the artificial basement membrane was significantly lower in two AG490 treatment groups than in control group（P0.05）.RT-PCR showed that the expression level of MMP-2 mRNA and VEGF mRNA was significantly lower in two AG490 treatment groups than in control group（P0.05）.Western blotting showed that the activation level of STAT3 protein and the expression level of MMP-2 and VEGF were lower in two AG490 treatment groups than in control group（P0.05）.Conclusion AG490 can down-regulate the expression of MMP-2 and VEGF by blocking the activation of STAT3,thus inhibiting the invasion of liver cancer SMMC-7721 cells in vitro.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第18期1960-1963,共4页
Journal of Third Military Medical University
