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为探讨Ph+白血病细胞中bcr/ab1表达的降低对细胞分化的影响,本文利用脂质体介导的方法将表达bcr/ab1融合区反义RNA片段的重组质粒导入K562和BV173细胞系,以Southern和Northern杂交以及筑巢式逆转录酶-聚合酶链反应技术证实外源DNA已在靶细胞内整合与表达,且bcr/ab1反义RNA片段的表达使内源性bcr/ab1 mRNA表达水平降低。未观察到表达bcr/ab1反义RNA片段的K562细胞出现粒单系或红系分化,重组质粒转染前后BV173细胞表面的CD10和CD34抗原以及K562细胞表面的CD13和CD33抗原表达无明显变化,提示bcr/ab1反义RNA片段抑制增殖的同时未引发分化与成熟。
In order to investigate the effect of the decrease of bcr/ab1 expression on cell differentiation in Ph+ leukemia cells, the recombinant plasmid expressing bcr/ab1 fusion region antisense RNA fragment was introduced into K562 and BV173 cell lines by liposome-mediated method. Northern hybridization and nested reverse transcriptase-polymerase chain reaction confirmed that the foreign DNA was integrated and expressed in the target cells, and the expression of bcr/ab1 antisense RNA fragment reduced the expression of endogenous bcr/ab1 mRNA. . No single-cell or erythroid differentiation was observed in K562 cells expressing bcr/ab1 antisense RNA fragments. The CD10 and CD34 antigens on the surface of BV173 cells and the expression of CD13 and CD33 antigens on K562 cells before and after recombinant plasmid transfection were not significantly changed. It was suggested that the bcr/ab1 antisense RNA fragment inhibited proliferation while not causing differentiation and maturation.