Bcl-2反义核酸增强HL-60、K562细胞对柔红霉素敏感性的研究  

Bcl-2 antisense oligodeoxynucleotide increases the sensitivity of HL60 and K562 cells to daunorubicin

作　　者：雷小勇[1] 张洹[1] 何冬梅[1] 

机构地区：[1]暨南大学医学院血液病研究所

出　　处：《中国病理生理杂志》2003年第2期194-197,T002,共5页Chinese Journal of Pathophysiology

基　　金：广东省科技计划重点基金资助项目 (99M0 1 2 0 4G);广州市科技计划重点基金资助项目 (2 0 0 1 -Z - 0 37- 0 1 )

摘　　要：目的 :研究bcl- 2反义寡核苷酸作用后 ,白血病细胞株HL60、K562细胞对柔红霉素 (DNR)敏感性的影响。方法 :MTT法测HL60、K562细胞中DNR的半数抑制率 (IC50 ) ;免疫荧光标记观测细胞Bcl - 2蛋白水平 ;用Hoechest332 58和碘化丙锭双染色法及流式细胞仪检测细胞凋亡。结果 :靶向bcl- 2mRNA蛋白编码区反义寡核苷酸(AS-ODN1 )和靶向翻译起始区的反义寡核苷酸 (AS -ODN2 )分别与DNR联合作用于HL60细胞后DNR的IC50值分别为 0 .1 2 4± 0 0 1 1、0 1 4 9± 0 0 1 2 ,分别与不加寡核苷酸组DNR的IC50值 (0 1 73± 0 0 2 1 )或无义寡核苷酸联用DNR的IC50值 (0 1 80± 0 0 2 3)相比有显著差异 ,P <0 0 5。AS -ODN1和AS -ODN2分别与DNR联合作用于K562细胞后DNR的IC50值分别为 0 0 78± 0 0 0 7、0 0 79± 0 0 0 8,分别与不加寡核苷酸组DNR的IC50值 (0 1 0 6± 0 0 1 1 )或无义寡核苷酸联用DNR的IC50值 (0 1 0 7± 0 0 1 2 )相比有显著差异 ,P <0 0 5。AS -ODN1和AS -ODN2分别与DNR同时作用于HL60或K562细胞后抑制Bcl- 2蛋白表达及诱导细胞凋亡率分别与单用DNR或无义寡核苷酸联用DNR相比有显著差异 ,P <0 0 5。与AS -ODN2比较 ,AS -ODN1提高HL60细胞对DNR的敏感性作用要强些 (P <0 0 5)。结论 :靶?AIM: To investigate whether the bcl-2 antisense oligonucleotide increases the sensitivity of HL60 and K562 cell lines to daunorubicin. METHODS: IC50 for HL60 and K562 was determined with MTT method, the expression levels of Bcl-2 protein were assayed by immunofluorescence using fluoresce isothiocyanate labeling. In addition, apoptosis was detected by morphological observation and flow cytometric analysis of DNA fragmentation. RESULTS: It was found that the two oligonucleotides directed against the coding region and the translation initiation of bcl-2 mRNA, combined respectively with daunorubicin, inhibited expression of bcl-2 protein, increased apoptosis in HL60 and K562 cells, and decreased IC50 of daunorubicin significantly ( P< 0.05). Compared to the antisense oligonucleotide directed against the translation initiation of bcl-2 mRNA, the antisense oligonucleotide directed against the coding region showed stronger effects in the aspects of increasing the sensitivity of HL60 cells to daunorubicin ( P< 0.05). CONCLUSIONS: These two antisense sequences in the translation initiation and the coding region of bcl-2 mRNA increased the sensitivity of HL60 and K562 cell lines to daunorubicin in a sequence-specific manner.

关 键 词：Bcl-2 反义寡核苷酸类 HL-60细胞 K562细胞 柔红霉素 药物敏感性 

分 类 号：R733.7[医药卫生—肿瘤]