摘要
目的探讨利用siRNA和靶向G3BP的多肽药物下调G3BP后对多种肿瘤细胞迁移能力的影响。方法采用人纤维肉瘤HT1080细胞、乳腺癌MCF-7细胞以及人非小细胞肺癌H1299细胞,应用siRNA特异性干扰G3BP后,通过划痕实验观察其对肿瘤细胞迁移能力的影响;用多肽药物GAP161作用于肿瘤细胞后,利用划痕实验以及Transwell迁移实验观察其对肿瘤细胞迁移能力的影响;在MCF-7细胞中高表达G3BP1,在MDA-MB-231细胞中用siRNA下调G3BP1后,采用人全基因芯片分析G3BP1高表达和低表达后的基因表达谱,并进行信号通路分析。结果 siRNA特异性下敲G3BP1和G3BP2后,HT1080、MCF-7和H1299细胞的迁移能力显著降低。利用靶向G3BP的特异性多肽药物GAP161作用于多种肿瘤细胞后,肿瘤细胞的迁移能力显著下调。全基因组芯片分析表明:多个基因同时在G3BP1高表达和低表达组中发生变化,该变化与细胞迁移相关的细胞黏附、整合素、MAPK等信号通路有关。结论 G3BP下调后能够显著降低肿瘤细胞的迁移能力。靶向G3BP的多肽药物具有显著抑制肿瘤细胞迁移的作用。下调或者高表达G3BP后可通过下调或者上调多种与细胞黏附、整合素、MAPK等信号通路相关基因发挥作用。
Objective To investigate the effect of G3BP down-regulation by siRNA or G3BP-targeted peptide GAP161 on tumor cell migration. Methods Effect of G3BP knockdown on tumor cell migration was examined by wound healing assay in HT1080, MCF-7 and H1299 cells. Effect of GAP161 on the migration of tumor cells was measured by wound healing assay and Transwell assay. Human whole genome oligonucleotide microarray was used to analyze the gene expression profiles in MCF-7 cells with G3BP1 over-expression and MDA-MB-231 cells with G3BP1 knockdown. Pathway analysis was then performed according to the microarray data. Results Down-regulation of G3BP1 and G3BP2 by specific siRNAs significantly inhibited cell migration in HT1080, MCF-7 and H1299 cells. GAP161 also significantly reduced the tumor cell migration ability. Whole genome microarray analysis showed that over-expression of G3BP1 and knockdown G3BP1 in MCF-7 and MDA-MB-231 cells, respectively, affected cell adhesion, integrin and MAPK signaling pathways in an opposite manner. Conclusion Knockdown of G3BP by siRNA significantly inhibited tumor cell migration. G3BP-targeted peptide G3BP1 markedly reduced the cell migration abilities. Over expression of G3BP1 could up-regulate genes expression associated with cell adhesion, integrin and MAPK signaling pathways, while down-regulation of G3BP1 exerts opposite effects.
出处
《中国医药生物技术》
2015年第1期11-17,共7页
Chinese Medicinal Biotechnology
基金
国家自然科学基金(81302802)
国家重点基础研究发展计划(973计划)(2009CB521807)
"重大新药创制"国家科技重大专项(2012ZX09301-002)
关键词
细胞系
肿瘤
细胞迁移抑制
基因表达谱
MAP激酶信号系统
G3BP
Cell line,tumor
Cell migration inhibition
Gene expression profiling
MAP kinase signaling system
G3BP
