RNA干扰抑制c-myc基因表达对D341细胞增殖的影响

目的探讨应用小干扰RNA（siRNA）技术抑制髓母细胞瘤D341细胞株的c-myc基因对D341细胞的生长、细胞周期及凋亡的变化。方法制备特异性针对c-myc基因的siRNA,转染D341细胞,实时定量RT-PCR、Western blot检测c-myc基因表达情况,流式细胞仪进行细胞周期及凋亡检测。结果siRNA转染D341细胞后,c-myc基因表达在转染后24h抑制效率最高,c-myc蛋白在48h抑制效率最高,并随siRNA浓度的增大,沉默c-myc基因效率增高。RNA干扰抑制D341细胞c-myc基因后,细胞生长停滞于G1期。结论针对c-myc基因的siRNA对D341细胞c-mycmRNA及蛋白有明显抑制作用,细胞生长受到抑制,有望成为髓母细胞瘤的新的治疗途径。

Inhibition of c-myc in Medulloblastoma Cell Line D341 by RNA Interference

Objective To investigate the inhibition of c-myc gene and protein in medulloblastoma cell line D341 by means of RNA interference in vitro,and investigate the cell cycle and apoptosis difference after interference. To explore the gene therapeutic possibility of siRNA for medulloblastoma. Methods The siRNA for c-myc gene was designed and transfected into D341 cells line. Real time PCR was carried out to detect the expression of target gene mRNAs,Immunoblots were carried out to detected the expression of c-myc protein. The cell cycle and apoptosis were detected by FCM. Results After transfection siRNA into D341 cells,c-myc gene was inhibited. C-myc gene was inhibited greatest at RNA level 24 hours after transfecting. At the 48th hour,there was the greatest inhibition effect for the protein level. With the increasing of concentration,the inhibition effect of c-myc gene raised. Silencing the targeted gene can induce the G1 phase arrest,which was confirmed by FCM. Conclusion The siRNA targeting c-myc gene can inhibit proliferation of D341 cells by inhibiting the expression of c-myc mRNA and protein and might be a new therapeutic modalities in treatment of medulloblastoma.