摘要
目的 研究 b3 a2型反义 bcr- abl寡核苷酸 ( ASO)体外对慢性髓细胞性白血病( CML )细胞株 K5 62的抑制作用 ,为反义技术用于 CML患者体内基因治疗和体外骨髓净化提供依据。方法 采用体外细胞培养技术及四唑盐 ( MTT)比色法、免疫组织化学染色法观察 b3 a2 - ASO体外对 K5 62细胞的生长、克隆形成及 P2 10 bcr- abl蛋白表达的影响。结果 经 b3 a2 - ASO( 110μg/ml)作用 4 0 h后 ,K5 62细胞生长抑制率达 66.12 % ,集落抑制率为 65 .4 4 % ;作用 15 h后 P2 10 bcr-abl蛋白合成抑制率可达 60 %。而无义寡核苷酸 ( NSO)对前述三个指标均无显著影响 ;b3 a2 - ASO及 NSO对 bcr- abl阴性细胞株 HL 60的细胞生长和存活率亦无显著影响。结论 b3 a2 - ASO对K5 62细胞有序列特异性抑制作用 ,提示其可以作为 CML反义基因治疗 ,尤其是骨髓净化的有力措施之一。
Objective To investigate the inhibition of K562 cells in vitro by antisense oligonucleotide targeted b3a2 typed bcr abl fusion gene and provide experimental supports for the utility of b3a2 ASO in the gene therapy and the bone marrow purging of the patients with chronic myelogenous leukemia (CML).Methods In vitro cell culture, MTT and immunohistochemical staining were used to observe the influences of b3a2 ASO on clonal formation, cell growth and expression of P210bcr abl protein. Results After 40 hour exposure to b3a2 ASO, the growth inhibition rate of K562 cells was 66.12% and the inhibition of clonal formation was 65.44%. The expression of P210bcr abl was reduced by 60% after 15 hour treatment. But nonsense oligonucleotide (NSO) had no pronounced influences on the cell growth and the clonal formation of K562 cells. In addition, neither b3a2 ASO nor NSO could significantly inhibit the cell growth and the clonal survival rate of bcr abl negative HL 60 cells. Conclusions b3a2 ASO had sequence specific inhibition to K562 cells, it could be used to gene therapy or bone marrow purging of the patients with CML .
出处
《临床内科杂志》
CAS
北大核心
2001年第6期422-424,共3页
Journal of Clinical Internal Medicine
基金
湖北省卫生厅资助项目
项目编号 :WT9741 5
关键词
慢性髓细胞性白血病
反义寡核苷酸
基因治疗
骨髓净化
Chronic myelogenous leukemia
Antisense oligonucleotide
Gene therapy
Bone marrow purging
