目的 研究GTP酶激活蛋白(Src同源结构域3)结合蛋白1[GTPase activating protein(Src homology domain 3)binding protein 1,G3BP1]是否在人非小细胞肺癌细胞系A549体外迁移中起作用,以判断该蛋白质是否可以作为抗肿瘤药物靶点.方法 人非小细胞肺癌细胞系A549及乳腺癌细胞系MDA-MB-231培养于10％ (V/V)的RPMI1640培养基中.以G3BP1过表达的乳腺癌细胞系MDA-MB-231作为阳性对照,采用Western印迹法检测A549细胞内G3BP1的表达情况;向A549细胞内转染以G3BP1的mRNA为靶标的小干扰RNA (siRNA)以抑制G3BP1的表达;分别检测G3BP1表达抑制后A549细胞穿过8 μm聚碳酸酯膜细胞数、定向运动距离及穿过人工基底膜细胞数.结果 Western印迹法显示G3BP1在非小细胞肺癌细胞系A549中过表达;转染siRNA后72 h内提取A549细胞蛋白质,Western印迹法检测显示G3BP1表达被完全抑制;G3BP1表达抑制后A549细胞穿过8μm聚碳酸酯膜细胞数、定向运动距离及穿过人工基底膜细胞数均显著减少(P＜0.05).结论 下调G3BP1表达抑制人肺癌细胞A549的体外迁移.%Objective To study the effect of GTPase activating protein (Src homology domain 3) binding protein 1 (G3BP1) on the in vitro migration of non-small cell lung cancer cell line A549, judging whether it can be designed as a antineoplastic drug target. Methods Human non-small cell lung cancer cell line A549 and breast cancer cell line MDA-MB-231 were cultured in RPMI 1640 containing 10% fetal bovine serum. Expression of G3BP1 in A549 was detected by Western blot with a positive control of MDA-MB-231 in which G3BP1 was overexpressed. Small interfering RNA(siRNA) targeted at the mRNA of G3BP1 was transfected into A549 cells to transiently knockdown G3BP1. Boyden chamber assay, wound healing assay, and in vitro invasion assay were performed to determine the amount of cells that passed 8 u,m polycarbonate membrane, directional migrationdistance and the number of cells that passed matrigel. Results Western blot showed an overexpression of G3BP1 in A549. Transient knockdown of G3BP1 inhibited the expression of G3BP1 in A549. The results showed great decrease in the number of cells that passed 8 μm polycarbonate membrane, directional migration distance and the number of cells that passed matrigel after knockdown of G3BP1 (P <0. 05). Conclusion Down-regulation of G3BP1 inhibits in vitro migration of human lung cancer cell A549.
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