【摘要】 目的　在三组具有不同转移潜能的癌细胞亚系中 ,在 RNA水平及蛋白水平检测 G3BP(Ras- GTPase- Ac-tiviting protein SH 3 dom ain binding protein)与肿瘤转移潜能的相关性。方法　研究对象为三组具有不同转移潜能的癌细胞亚系 ,即 PG(人肺巨细胞癌 ,裸鼠体内转移率 10 0 % )与PAa(人肺腺癌 ,无转移 ) ;PG亚系 BE1与 L H 7(前者裸鼠体内转移率 10 0 % ,后者无转移 ) ;1E8与 2 B4来自人前列腺癌PC- 3M(前者裸鼠体内转移率 10 0 % ,后者无转移 )。检测方法包括 Northern杂交 ,蛋白质 Western印迹免疫化学 ,免疫组织化学。结果　应用 m RNA差异显示技术对比研究具有不同转移潜能的亚系 BE1,L H 7时 ,发现一个 849bp片段在BE1低表达 ,而在 L H7高表达 ,经 Northern杂交验证其表达差异后测序 ,Gen Bank Blastn检索 ,与 G3BP同源性高达99%。在三组具有不同转移潜能的癌系中 ,Northern杂交 ,蛋白质 Western印迹免疫化学显示 G3BP在低转移癌系L H7,2 B4,PAa中表达显著高于其所对应的高转移癌系BE1,1E8,PG。免疫组织化学提示 G3BP蛋白位于胞质 ,低转移癌系 L H7,2 B4,PAa表达显著高于其所对应的高转移癌系 BE1,1E8,PG(PG各细胞染色不均一 )。结论　在 m R-NA水平和蛋白质水平 ,G3BP在三组高低转移癌系中的表达具?更多还原

【Abstract】 Objective The relationship between G3BP (Ras GTPase Activating protein SH3 domain binding protein) and cancer metastasis was studied. Methods G3BP expression in three pairs of cancer cell sublines with/without metastatic potential (BE1/LH7, 1E8/2B4 and PG/PAa) was confirmed by Northern hybridization, Western blotting and immunohistochemical analysis. Results G3BP was obtaine by mRNA differential display and it displayed a strong expression in LH7 (no metastatic potential in recipient nude mice ) and a very weak expression in BE1 (100% metastatic frequency). Northern blotting and Western blotting with a polyclonal G3BP antibody confirmed an increased G3BP expression in non metastatic cell lines (LH7, 2B4, PAa) compared with highly metastatic cell lines (BE1, 1E8, PG). Immunohistochemical analysis demonstrated a prominent granular pattern of G3BP accumulation in cytoplasm. Conclusion The results revealed a significant association between G3BP expression and cancer metastasis, due to its different expressions in messenger RNA and protein level between the three pairs of cell sublines with different metastatic potentials.更多还原