摘要
目的观察抑制容积调控氯通道ClC-2基因的表达对胶质瘤BT-325细胞生长的影响。方法设计和构建两个针对ClC2基因的小干扰RNA(siRNA)重组表达载体,用脂质体LipofectamineTM2000介导转染,将空载体质粒和两个重组质粒分别转染入BT-325细胞(依次为对照组、PP1组和PP2组);通过RTPCR检测ClC2基因mRNA表达变化;MTT分析检测细胞活性;流式细胞仪检测细胞周期。结果与对照组相比较,干扰组ClC2基因的mRNA水平明显降低,细胞生长速度明显减慢,细胞周期进程被阻滞在G1期。结论干扰胶质瘤细胞系BT-325细胞的ClC-2基因表达可以抑制细胞的生长,提示ClC2基因可能成为控制胶质瘤恶性生长的新靶点。
Objective To observe the growth of BT-325 human glioma cells after interfering volume regulated chloride channel ClC-2 gene. Methods Two expression recombinant vectors of ClC-2 gene were designed and constructed. The primary plasmid, pSUPER. puro-shRNA, and the two recombinant plasmids, pSUPER, puro-shRNA-ClC-21 and pSUPER, puro-shRNA-ClC-22, were transfected into BT-325 cells by Lipofectamine^TM 2000 （Groups： control, PP1 and PP2, respectively）. The mRNA expression of ClC-2 gene was detected with reverse transcription polymerasse chain reaction （RT-PCR）, the cellular survival was determined with MTT assay, and the cell cycle was measured with flow cytometry （FCM）. Results ClC-2 mRNA expression and the growth of the cells in PP1 and PP2 groups were significantly lower than that of control group. The cell cycle progression was blocked in G1 phase （PP1 and PP2 vs control, P 〈 0.01）. Conclusion The growth of BT-325 human glioma cells is prevented by knockdown of ClC-2 gene expression, which may be one of the novel targets to inhibit growth of human malignant glioma cells.
出处
《中国康复理论与实践》
CSCD
2006年第5期378-380,共3页
Chinese Journal of Rehabilitation Theory and Practice
基金
国家自然科学基金资助项目(30270602)。
