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目的:通过脂质体分别转染bcl-2两个靶点的反义寡核苷酸(ASODN1、ASODN2)及G3139,观察bcl-2反义寡核苷酸对淋巴瘤细胞株Raji凋亡的影响。方法:采用MTT法检测药物的半数抑制率(IC50);免疫荧光标记观察细胞bcl-2蛋白水平;用姬姆萨染色和流式细胞仪观察细胞凋亡。结果:MTT测定显示:ASODN1、ASODN2组IC50值分别与G3139组进行比较无显著差异,P>0.05。5μmol/LASODN1、ASODN2作用于Raji细胞72h后,姬姆萨染色均可见凋亡细胞。5μmol/LASODN1、ASODN2和G3139作用于Raji细胞后,bcl-2蛋白阳性率均明显下降,细胞凋亡率均明显增加,P<0.05,但ASODN1、ASODN2组分别与G3139组进行比较无显著差异,P>0.05。5μmol/L无义寡核苷酸对Raji细胞的生长活性、bcl-2蛋白水平及细胞凋亡率均无明显影响,P>0.05。结论:针对bcl-2两个靶点的反义寡核苷酸能特异性促进Raji细胞的凋亡,并且与G3139具有近似的反义效应。
OBJECTIVE: To observe the effect of bcl-2 antisense oligonucleotide on the apoptosis of Raji cell line Raji via antisense oligonucleotide (ASODN1, ASODN2) and G3139 transfected by bcl-2 targeting liposome respectively influences. Methods: The half inhibition rate (IC50) of the drug was detected by MTT assay. The bcl-2 protein level was observed by immunofluorescence staining. Apoptosis was observed by Giemsa staining and flow cytometry. Results: MTT assay showed that there was no significant difference between IC50 values of ASODN1 and ASODN2 group and G3139 group respectively. After treated with ASODN2 or ASODN2 for 72h, apoptotic cells were found in Giemsa staining. After treated with 5μmol / L ASODN1, ASODN2 and G3139, the positive rate of bcl-2 protein was significantly decreased, and the apoptosis rate was significantly increased (P <0.05). However, there was no significant difference between ASODN1 and ASODN2 groups and G3139 group, P> 0.05.5μmol / L nonsense oligonucleotide had no significant effect on the growth activity, bcl-2 protein level and apoptosis rate of Raji cells, P> 0.05. CONCLUSION: Antisense oligonucleotides directed against two targets of bcl-2 specifically promote apoptosis of Raji cells and have approximate antisense effects to G3139.