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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。
Objective To investigate the effect of small interfering RNA (siRNA) on the growth, cell cycle and apoptosis of D341 cells induced by c-myc gene in D341 cell line. Methods siRNA targeting c-myc gene was transfected into D341 cells. The expression of c-myc gene was detected by RT-PCR and Western blot. The cell cycle and apoptosis were detected by flow cytometry. Results After transfected with D341 cells, the expression of c-myc gene was the highest at 24h after transfection. The efficiency of c-myc protein was highest at 48h after transfection. The efficiency of silencing c-myc gene increased with the increase of siRNA concentration. After RNA interference inhibited c-myc gene in D341 cells, cell growth arrest in G1 phase. Conclusions siRNA targeting c-myc gene can significantly inhibit c-myc mRNA and protein expression in D341 cells and inhibit the growth of cells. It is expected to be a new therapeutic approach for medulloblastoma.