bcl-2基因反义寡核苷酸增强放射线诱导Raji细胞凋亡的研究

The study on bcl-2 antisense oligodeoxynucleotide increasing radiation-induced apoptosis in Raji cell line

目的研究bcl-2基因反义寡核苷酸作用淋巴瘤细胞株Raji后,放射线对细胞凋亡的影响。方法bcl-2基因反义寡核苷酸作用Raji细胞后,台盼蓝拒染法计数活细胞;免疫荧光标记观测细胞Bcl-2蛋白水平;用姬姆萨染色和碘化丙啶染色法、流式细胞仪检测细胞凋亡。结果bcl-2基因反义寡核苷酸与放射线联合作用Raji细胞,显著抑制细胞的生长,并呈时间依赖性,活细胞数分别与单用放射线组及无义寡核苷酸联合放射线组相比,统计学上有显著性差异(P<0.05)。bcl-2基因反义寡核苷酸与放射线联合作用于Raji细胞后显著抑制bcl-2蛋白表达,蛋白水平分别与单用放射线组或无义寡核苷酸联用放射线组比较,差异有显著性(P<0.05)。bcl-2基因反义寡核苷酸与放射线同时作用于Raji细胞后,姬姆萨染色可见凋亡细胞,通过流式细胞仪检测的细胞凋亡率显著增加,分别与单用放射线组或无义寡核苷酸联用放射线组相比,具有显著性差异,P<0.05。结论bcl-2反义寡核苷酸能增强放射线诱导Raji细胞凋亡。

Objective To investigate whether the bcl-2 antisense oligonucleotide（ASODN） could increase radiation-induced apoptosis in Raji cell line. Methods The surviving cells were determined using the trypan blue dye exclusion assay. The expression levels of Bcl-2 protein were assayed by immunofluorescenced using fluoresce isothiocyanate label. Apoptosis was detected by Giemsa staining and flow cytometric cell cycle analysis. Results It was found that bcl-2 ASODN combined with radiation had significantly reduced the number of viable cells （P〈0.05）. There was no difference in cells survival between nonsense oligodeoxynucleotide （NSODN）/radiation combination and radiation-treated cells alone, bcl-2 ASODN combined radiation could significantly inhibit expression of Bcl-2 protein in Raji cells （P〈0.05）. In morphological observation of apoptotic cells using Giemsa staining, ceils treated with bcl-2 ASODN combined with radiation at 72 hours displayed classic apoptotic changes. Apoptosis rate of Raji cells treated with bcl-2 ASODN combined with radiation significantly increased （P〈0.05）, compared with NSODN/radiation combination and radiation -treated cells alone, respectively. Conclusion bcl-2 antisense oligonucleotide can increase radiation-induced apoptosis in Raji cell line.