bcl-2硫代反义寡核苷酸对Raji细胞增殖和凋亡的影响

研究bcl-2硫代反义寡核苷酸(bcl-2 ASODN)对淋巴系统恶性肿瘤的作用及机制。将bcl-2 ASODN与Raji细胞共孵育，应用MTT检测，电镜超微结构，流式细胞仪（FCM）分析和RT-PCR方法，观察bcl-2 ASODN对Raji细胞增殖及凋亡的影响，以及bcl-2蛋白及mRNA水平的变化。MTT检测显示bcl-2 ASODN可以部分抑制Raji细胞增殖；经ASODN作用48h，电镜超微细胞观察细胞呈现典型凋亡特征，包括胞膜出芽，异染色质核膜下浓聚形成凋亡小体及胞核破裂；FCM检测Annexin V^ /PI^-的凋亡细胞比例升高，20μmol/L bcl-2 ASODN处理Raji细胞72h，细胞凋亡率达43.86%，比对照组（10.05%）明显增高。bcl-2阳性细胞率逐渐下降，于72h达低谷为0.88%，明显低于对照组（79.54%）。bcl-2 mRNA水平亦显下降。结论 bcl-2 ASODN通过下调bcl-2蛋白表达水平，诱导Raji细胞凋亡。

Effects of bcl-2 antisense oligodeoxynucleotide on proliferation and apoptosis of Raji cells

OBJECTIVE: To study the in vitro antitumor activity of bcl-2 fully phosporothioated antisense oligodeoxynucleotide (bcl-2 ASODN) to malignant lymphoblastic cells. METHODS: Proliferation and apoptosis of Raji cells incubated with bcl-2 ASODN were evaluated by MTT assay, flow cytometry (FCM) and electron microscopy, and the level of bcl-2 protein and mRNA expression were assessed by FCM and RT-PCR, respectively. RESULTS: MTT assay demonstrated that bcl-2 ASODN could partially inhibit the growth of Raji cells. After incubated with ASODN for 48 hours, Raji cells exhibited characteristic morphologic changes of apoptosis, including cytoplasm membrane blebbing, chromatin condensation crescents formation and nuclear fragmentation. The apoptosis rate of Raji cells treated with 20 micromol/L bcl-2 ASON for 72 hrs was 43.86% which is significantly higher than that of control (10.05%). The bcl-2 ASODN induced apoptosis of Raji cells was accompanied by declined expression of bcl-2 mRNA, which decreased to 0.88% at 72 hrs and was significantly lower than that of control (79.54%). CONCLUSION: bcl-2 ASODN induced Raji cells apoptosis by downregulating bcl-2 protein.