摘要
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 explore the effect of liposome transfection antisense phosphorothioate oligodeoxynucleotide directed against the coding region of the bcl-2 messenger RNA and the translation site, and G3139 on apoptosis in Raji cells. Methods: Cytotoxic effects were measured by use of MTT method; The expression levels of bcl-2 protein were assayed by immunofluorescence using fluoresce isothiocyanate label. Apoptosis was determined by morphological observation and flow cytometric analysis. Results: The two antisense oligonucleotides and G3139 can reduce bcl-2 protein levels and Raji cells viability (IC50=4.54, 4.72 and 4.26靘ol/L, respectively), and induce apoptosis. The scrambled sequence control oligonucleotides and empty liposomes did not alter cell viability? bcl-2 protein expression and apoptosis rates. There was no difference in reducing bcl-2 protein levels and apoptosis rates induced among these three antisense oligonucleotides. Conclusions: The two antisense oligodeoxynucleotide of bcl-2 messenger RNA can effectively induce apoptosis of Raji cells. The two antisense sequence and G3139 have similarity antisense effect.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2004年第12期664-666,共3页
Chinese Journal of Clinical Oncology
基金
广东省自然科学基金(编号:021195)
广州市科技计划重点基金资助(编号:2001-Z-037-01)
