bcr/abl反义寡聚脱氧核苷酸抑制慢性粒细胞白血病细胞生长的研究

Inhibitory effects of bcr/abl antisense oligodeoxynucleotide on chronic myelocytic leukemic cell growth

目的:研究bcr/ abl基因的反义寡聚脱氧核苷酸(antisense oligodeoxynucleotide,ASON)对K5 6 2细胞生长的影响。方法:根据bcr/ abl基因类型(b3/ a2 )的c DNA序列分析,以其m RNA融合区的18个核苷酸为作用靶序列,人工合成了18聚ASON片段,同时合成18聚无义ASON片段,作为序列特异性对照。用ASON和无义ASON片段与K5 6 2细胞共同培养,同时设置空白对照组,作用一定时间后分别测定细胞动力学、3H- Td R掺入率、bcr/ abl m RNA和P2 10蛋白的表达。结果:3.5～30 .0 μmol·L- 1的ASON对K5 6 2细胞生长有不同程度的抑制作用,ASON浓度低于10 .0 μmol·L- 1时抑制作用很小,大于10 .0 μm ol·L- 1时抑制作用显著,这种作用有浓度、时间依赖关系,而无义ASON与空白对照组比较差异无显著性(P<0 .0 5 )。同时发现ASON使细胞的3H - Td R掺入率下降,bcr/ abl m RNA及P2 10蛋白的表达下降。结论:ASON的抑制作用是从基因水平上封闭了bcr/ abl m RNA转录及P2 10蛋白的翻译,抑制细胞的增殖,促进细胞凋亡。ASON对慢性粒细胞白血病(CML )细胞有抑制作用。

Objective To study the effects of bcr/abl antisense oligodeoxynucleotide (ASON) on K562 cell growth. Methods Eighteen nucleotides were used as target genes in bcr/abl mRNA mixed area, by different bcr/abl cDNA sequence type analysis, 18 pdy ASON fragments and 18 poly non-sense ASON fragments for special sequence control were synthesized. Incubating K562 cells with ASON or non-sense ASON fragments for a mean time, while a blank control was set up. Cytomorphology, the 3H-TdR mixed rate and the expressions of bcr/abl mRNA and P210 protein were detected. Results ASON had different inhibitory rates on cell proliferation at 3.5- 30.0 μmol·L -1 . The action was smaller when ASON concentration was less than 10.0 μmol·L -1 . While the action was more significant when the concentration was larger than 10.0 μmol·L -1 . This action depends on concentration and time. But non-sense ASON and control weren′t distinct different. At the meantime, ASON made the 3H-TdR mixed rate decrease and the expression levels of bcr/abl and P210 were down-regulated. Conclusion These results show that ASON hinders bcr/abl mRNA transcription and P210 protein translation, inhibits cell proliferation and promotes apoptosis on gene levels.