摘要
目的 探讨CD40基因特异性沉默对脑胶质瘤细胞增殖和迁移等恶性生物学行为的影响.方法 构建特异性沉默CD40基因的重组表达载体(pSilencer3.1/shRNA-CD40)和随机对照载体(pSilencer3.1/shRNA-controi),转染脑胶质瘤细胞株U87.采用RT-PCR和流式细胞术分别在mRNA和蛋白水平的检测CD40基因沉默效率,进而采用细胞计数法和Transwell法观察下调CD40分子表达后的U87细胞在sCD40L作用下增殖能力和迁移能力的改变.结果 成功构建了特异性沉默CD40基因的稳定细胞株U87/shRNA-CD40,可显著下调U87细胞CD40 mRNA 和蛋白质表达水平(P＜0.05).采用sCD40L激发CD40信号,结果显示,与野生型U87和对照组U87/shRNA-control相比,U87/shRNA-CD40细胞株增殖速率和迁移能力明显下降(均P＜0.01).结论 特异性沉默脑胶质瘤细胞CD40分子表达,可以有效削弱肿瘤微环境中CD40信号导致的脑胶质瘤细胞增殖和迁移能力,这为临床治疗脑胶质瘤提供了新的线索.
Abstract
Objective To discuss the effect of CD40 gene silencing on proliferation and migration of glioma cells. Methods The plasmids pSilencer3. L/shRNA-CD40 targeting CD40 gene and pSilencer3. 1/shRNA-control were constructed and then transfected into glioma cell line U87. The efficiency of CD40 silencing was evaluated with RT-PCR and FACS. The proliferation and migration ability of U87 stimulated with sCD40L were analyzed by cell counting and transwell assay. Results Expression plasmid (pSilenc-er3. L/shRNA-CD40) against CD40 siRNA could downregulate the CD40 gene and protein expression dramatically. Compared with U87 and the U87/shRNA-control groups, the proliferation and migration ability of U87/shRNA-CD40 cells were decreased under stimulation of sCD40L( P < 0. 01). Conclusion The siRNA against CD40 gene could significantly inhibit proliferation and migration ability of glioma cell line U87 providing a clue for molecular targeting treatment of glioma.
NETL
NSTL
维普
万方数据