摘要
目的研究无血清状态下,FBJ-S1细胞较FBJ-LL细胞更易死亡的原因。方法用细胞活性检测法分析细胞的活性,分别用HPTLC法、逆转录PCR法和免疫印迹法检测细胞中GD1a和窖蛋白1(caveolin-1)的含量。结果研究表明,在无血清培养基培养条件下,富含神经节苷脂GM3和GD1a的低转移性的FBJ-S1小鼠骨肉瘤细胞趋向死亡,但对含有几乎相同量GM3而无GD1a表达的高转移性的FBJ-LL细胞的生长却未见明显影响。同时发现:a.FBJ-S1细胞富含窖蛋白1,而FBJ-LL细胞中窖蛋白1表达量则较低;b.利用小干扰基因沉默技术在FBJ-S1细胞中沉默窖蛋白1的表达,结果使细胞存活率升高;c.使用GD1a处理FBJ-LL细胞导致其趋向死亡,通过免疫印迹法检测窖蛋白1发现其蛋白表达量增加。结论在无血清培养条件下,窖蛋白1诱导FBJ细胞的死亡。
Objective To investigate the reason why FBJ-S1 but not FBJ-LL cells die in serum-free medium.Methods Cells viability was detected by WST assay. The expression of GDla and caveolin-1 (Carl) was monitored by HPTLC and RT-PCR or Western blots, respectively. Results Poorly metastatic mouse osteosarcoma FBJ-S1 cells expressing gangliosides GM3 and GDla underwent cell death, but highly metastatic FBJ-LL cells containing the same GM3 content though only very little GDla did not. In addition, 1) FBJ-S1 cells were rich in Carl compared to FBJ2 LL cells, 2) Cavl siRNA transfected S1 cells survived scrambled sequence transfected S1 cells, and 3) cultivation of FBJ-LL cells with GDla resulted in cell death in serum-free culture with elevated Carl expression as revealed by Western blotting. Conclusions Caveolin-1 is responsible for cell death under serum-free conditions.
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2008年第3期223-228,共6页
Journal of Shenyang Pharmaceutical University