RNA干扰Fascin1表达抑制胶质瘤细胞U87MG的侵袭、转移能力

Reversal effect of RNA interference targeting Fascin1 on migration and invasion of glioma cell line U87 MG

目的:本研究运用RNA干扰(RNA interference,RNAi)技术特异性抑制肌动蛋白结合的细胞骨架蛋白Fascin1的表达,并探讨其对胶质瘤细胞U87 MG的侵袭和转移能力的影响及相关分子机制。方法:将针对Fascin1的小干扰RNA(Fascin1-si RNA)和阴性对照si RNA(Negative-si RNA)转染至人胶质瘤细胞株U87 MG中。实验分为3组:空白对照组(未转染的细胞)、阴性对照组(转染neg-si RNA的细胞)、实验组(转染Fascin1-si RNA的细胞)。用transwell小室法检测Fascin1对U87MG细胞的侵袭和转移能力的影响,蛋白印迹法(Western blot)检测U87 MG细胞中Fascin1、p AKT及p STAT3等蛋白的表达。通过添加针对PI3K通路的抑制剂LY294002和针对STAT3通路的抑制剂JSI-124,证实了这两条信号通路在胶质瘤细胞U87MG侵袭、转移方面的作用。结果:相比空白对照组和阴性对照组,实验组细胞的转移能力降低了约(49±5.8)%及(46±5.4)%(P<0.05),侵袭能力降低了约(42±4.9)%和(43±4.8)%(P<0.05);且相关的PI3K/AKT及STAT3等信号通路的磷酸化减少;添加相应的信号通路抑制剂后,细胞的侵袭、转移能力显著降低(P<0.05)。结论:RNAi抑制Fascin1表达能够降低胶质瘤细胞U87 MG的侵袭及转移能力,并可抑制PI3K/AKT和STAT3信号激活,在一定程度上可逆转胶质瘤细胞的转移和侵袭等肿瘤特性。

Objective： To detect the functional role of Fascin1 and its related molecular mechanisms in migration and invasion capacity of glioma cells,we utilized gene specific small interference RNA of Fascin1 in cell line U87 MG. Methods： Fascin1-si RNA or negative si RNA was transfected into U87 MG cells of control group or experiment group. Transwell method was employed to assess the migration and invasion capacity of glioma cells. Western blot analysis was used to detect the protein expression of Fascin1,p AKT and p STAT3. The impact of PI3 K / AKT pathway and STAT3 pathway on migration and invasion of U87 MG cells was verified,via applying LY294002 and LY294002,which was inhibitor of the two pathways respectively. Results： As compared to control groups,the migration and invasion capacity of transfected glioma cells were attenuated about 52% or 43%（ P〈0. 05）,accompanied with the decreased phosphorylation of AKT and STAT3. As utilizing the inhibitors of AKT and STAT3,attenuated migration and invasion capacity of U87 MG cells were observed. Conclusion： Down-regulated expression of Fascin1 could suppress the migration and invasion capacity of U87 MG cells by inhibiting the phosphorylation of PI3 K / AKT pathway and STAT3 pathway.