摘要
目的:观察人慢性粒细胞白血病(CML)细胞转导抗ABL胞内抗体基因后,对其酪氨酸激酶活性的影响。方法:构建逆转录病毒载体MSCV-ib-IRES-eGFP,转导K562细胞,分选eGFP+的细胞,用RT-PCR检测胞内抗体mRNA的表达,观察细胞内BCR/ABL、c-ABL酪氨酸激酶活性及细胞总蛋白酪氨酸激酶活性的变化。结果:获得表达胞内抗体的K562细胞:K562-ib-eGFP。RT-PCR证实胞内抗体mRNA在K562-ib-eGFP细胞内表达。与对照组相比,K562-ib-eGFP细胞内BCR/ABL和c-ABL蛋白酪氨酸激酶活性及总蛋白酪氨酸激酶活性明显受抑制。结论:转导抗ABL胞内抗体能有效降低CML细胞内ABL酪氨酸激酶活性,为进一步研究打下基础。
Objective： To study the effects of anti-ABL intrabody on the tyrosine kinase activity of human chronic myelogenous leukemia （CML） cells in vitro. Methods：A recombinant retroviral vector MSCV-ib-IRES-eGFP was constructed. K562 cells were transduced with the retrovirus and eGFP^＋ cells were selected. The intrabody mRNA expression was determined by RT-PCR. BCR/ABL and c-ABL protein tyrosine kinase （PTK） activity and the whole cell PTK activity were examined. Results： An intrabody expressing K562 cell line, K562-ib-eGFP ,was obtained. Expression of the intrabody mRNA in K562-ib-eGFP cells was confirmed by RT-PCR. BCR/ABL and c-ABL protein tyrosine kinase （PTK）activity and the whole cell PTK activity of K562-ib-eGFP cells were significantly inhibited as compared with those of control groups.Conclusion：Transduction of CML cells with the intrabody containing retrovirus efficiently inhibits the ABL protein tyrosine kinase activity, which may be helpful to further studies for CML.
出处
《天津医科大学学报》
2007年第4期539-542,共4页
Journal of Tianjin Medical University
