摘要
构建了表达2、3和4个相同241bp b3/a2型bcr/abl融合基因的反义RNA片段的重组质粒,酶切电泳和以241bp bcr/abl基因片段为探针进行的菌落原位杂交证实各目的片段已插入逆转录病毒表达载体,分别命名为pDAB2、pDAB3和pDAB4。用脂质体介导的方法将pDAB3导入Ph^+人白血病细胞系(K562和BV173细胞),48小时后用G418进行抗性筛选,观察了对靶细胞增殖的影响。结果表明,外源质粒在靶细胞内表达的反义RNA片段,使细胞致瘤性消失及诱发凋亡。探讨了可能的作用机理。
The recombinant plasmids in which two, three and four identical 241 bp ber/ abl (b3/ a2 type) fu-sion gene fragments vvere cloned into pDORneo vector in antisense orientation were constructed. Electrophoresis following proper enzymatic digestions and colony in situ hvbridization with 241 bp ber/abl fragment as probe proved the correct insertion of bcl/ abi fragments. The plasmid pDAB3 was transfected into Ph+ human leukemia celi lines K562 and BV173 with lipofectin reagent. G418 was given 48 hours later and the effects on cell proliferation were observed. The results indicated that the expression of antisense transcripts to ber/abl fusion gene caused apoptosis disappearance of carcinogenicity. The possible mechanism was dis-cussed.
出处
《中国实验血液学杂志》
CAS
CSCD
1996年第1期46-55,共10页
Journal of Experimental Hematology
