摘要
目的观察用AG490阻断人胶质瘤细胞株中信号传导和转录活化因子3（STAT3）信号通路对肿瘤细胞侵袭性特征的影响。方法体外培养的人胶质瘤U251、U87细胞株，应用AG490阻断STAT3信号通路；以Western blot检测STAT3和其激活状态p-STAT3蛋白在AG490作用前后的表达情况；通过Transwell细胞侵袭实验了解肿瘤细胞体外侵袭能力的变化；用明胶酶谱法检测肿瘤细胞基质金属蛋白酶（MMP）-2、MMP-9的表达；用逆转录-聚合酶链反应（RT-PCR）了解血管内皮生长因子（VEGF）mRNA表达的变化。结果AG490作用后胶质瘤细胞内p-STAT3表达下降，其程度随AG490浓度升高而增强，差异有统计学意义（P〈0．01）。而STAT3蛋白的表达不受AG490影响，与对照组比较差异无统计意义（P〉0．05），说明AG490的作用机制是通过抑制STAT3蛋白的磷酸化激活过程从而阻断STAT3信号通路。AG490作用后，U251细胞的体外侵袭力下降，差异有统计学意义（P〈0．01）。STAT3信号通路阻断后，胶质瘤细胞MMP-2、MMP-9的表达下凋，差异有统计学意义（P〈0．01）。VEGF mRNA表达也相应下降（P〈0．05）。结论应用AG490阻断STAT3信号通路能够抑制胶质瘤细胞的体外侵袭能力，STAT3信号通路有可能成为控制胶质瘤侵袭性生长的有效靶点。
Objective To study the role of STAT3 signal pathway in the invasiveness of human glioma cell lines. Methods The STAT3 signal pathway was blocked in glioma cell lines U251 and U87 by AG490. Western blot was applied to detect the inhibition of STAT3 signal pathway. Transwell cell invasive assay was performed to observe the effect of AG490 on the glioma invasiveness in vitro. The expression of MMP-2 and MMP-9 was assayed by Zymography. RT-PCR was used to detect the expression of VEGF mRNA. Results The expression of p-STAT3 was significantly inhibited in a concentration-dependent manner in U251 and U87 cell lines after AG490 treatment （ P 〈 0.01 ） ,but that of STAT3 stayed unchanged （ P 〉 0.05）. Transwell cell invasive assay showed that the invasiveness of glioma cells was significantly decreased after the AG490 treatment （ P 〈 0.01 ）. And the expression of MMP-2 and MMP-9 was down-regulated after the blocking of the STAT3 signal pathway （ P 〈 0.01 ）. Meanwhile, a decrease of VEGF mRNA expression was also observed by RT-PCR （ P 〈 0.05）. Conclusion STAT3 signal pathway played an important role in the invasiveness of glioma cell lines. It was expected to be a promising target in the treatment of glioma.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2008年第8期956-958,共3页
Chinese Journal of Experimental Surgery
