Survivin-siRNA对骨肉瘤MG-63细胞药物敏感性的研究

Effect of Survivin-siRNA on the drug sensitivity of osteosarcoma cell line MG-63

目的研究特异性Survivin-siRNA转染骨肉瘤MG-63细胞后对骨肉瘤细胞药物敏感性的影响。方法应用两种特异性Survivin-siRNA(Survivin-siRNA1,Survivin-siRNA2)转染骨肉瘤MG-63细胞,RT-PCR检测MG-63细胞Survivin-mRNA表达,流式细胞术检测MG-63细胞的凋亡,MTT法检测顺铂、多柔比星对转染前后MG-63细胞的半数抑制浓度(IC50)。结果特异性Survivin-siRNA转染骨肉瘤MG-63细胞24h后SurvivinmRNA表达水平明显抑制;流式细胞术显示转染Survivin-siRNA的MG-63细胞48h后凋亡率明显高于非特异性siRNA组和空白对照组(F=17.32,P<0.01);特异性Survivin-siRNA增强了MG-63细胞对多柔比星的敏感性5倍,增强了MG-63细胞对顺铂的敏感性9倍。结论化学合成的Survivin-siRNA能有效抑制Survivin基因在骨肉瘤MG-63细胞中的表达,并提高MG-63细胞对多柔比星、顺铂的敏感性。

Objective To explore the effect of Survivin-siRNA on the sensitivity of osteosarcoma cell line MG-63. Methods Two survivin siRNAs （siRNA~, siRNA2） which are the specifically targeted survivin genes were chemically synthesized and transfected into MG-63 cells. Survivin mRNA expression was detected by reverse transcription-polymerase chain reaction （RT- PCR）. Apoptosis rate was analyzed by flow cytometry （FCM）, and 50% inhibition concentration （IC50） of adriamycin （ADM） and diamminedichloroplantinum （DDP） on MG-63 cells was determined with the MTT method. Results The expression level of survivin mRNA was significantly inhibited 24 hours after MG-63 cells were transfected with siRNA. Cytometry showed that the apoptosis rate of MG-63 cells was significantly higher in survivin-siRNA group than in nonspecific siRNA and empty vector group 48 hours after transition with survivin-siRNA （23.7% ± 1.5%, 25.8% ± 2.3%, 7.6% ± 0.7%, or 5.2% ± 0.3%, F=17.32, P〈0.01）. Specific survivin-siRNA increased 5 folds of the sensitivity of MG-63 cells to ADM and 9 folds to DDP. Coneluslon Chemically synthesized short survivin-siRNA can effectively inhibit the expression of survivin gene in osteosarcoma MG-63 cells and enhance the sensitivity of MG-63 cells to ADM and DDP.