摘要
目的观察人慢性粒细胞白血病细胞转导抗ＡＢＬ蛋白酪氨酸激酶区胞内抗体基因后，对其在裸鼠体内生长的影响。方法应用基因重组技术构建逆转录病毒载体ＭＳＣＶ－ｉｂＥ－ＩＲＥＳ－ｅＧＦＰ，将胞内抗体基因转导Ｋ５６２细胞，观察胞内抗体的表达和细胞内ｃ－ＡＢＬ及ＢＣＲ／ＡＢＬ蛋白酪氨酸激酶活性的变化；将表达胞内抗体的细胞与对照组Ｋ５６２细胞及转导空载体的细胞分别移植裸鼠，动态观察肿瘤的生长。结果获得表达胞内抗体基因的细胞模型：Ｋ５６２－ｉｂＥ，其靶蛋白酪氨酸激酶活性明显受抑。移植裸鼠后第１４，２１，２８天肿瘤体积均小于对照组的１／２。结论转导抗ＡＢＬ蛋白酪氨酸激酶区胞内抗体的细胞在裸鼠体内生长明显受抑制，这可能与胞内抗体显著抑制细胞内ＢＣＲ／ＡＢＬ蛋白酪氨酸激酶活性，阻断ＢＣＲ／ＡＢＬ信号途径，促进细胞凋亡，降低了细胞的体内致肿瘤性有关。
Objective To study the effects of anti-ABL tyrosine kinase intrabody on the growth of human chronic myelogenous leukemia(CML)cells in nude mice.Methods A recombinant retroviral vector MS -CV-ibE-IRES-eGFP was constructed to express intracellular single-chain antibody(intrabody)against ABL tyrosine kinase domain in CML cells.K562cells were transduced with the retrovirus,eGFP +cells were then selected by fluorescence-activated cell sorting(FACS).The intrabody mRNA expression was determined by reverse transcription(RT)-polymerase chain reaction(PCR).BCR/ABL and c-ABL protein tyrosine kinase(PTK)activity in the cells was examined.Transduced cells and control group K562cells were transplanted into nude mice respectively and the tumor sizes were dynamically obse r-ved.Results K562-ibE cell was obtained.Expression of the BCR/ABL and c-ABL protein tyrosine kinase activity of harvested K562-ibE cells were markedly inhibited.At 14,21and28days after cell injection,the tumor volumes of experimental mice were obviously smaller than that of control mice,about one half of the control groups(P<0.05=.Conclusion The growth of K562-ibE cells was sign-ificantly inhibited in vivo.It is possible that inhibition of the BCR/ABL protein tyrosine kinase act-ivity by the intrabody blocked BCR/ABL signal transduction pathway,promoted apoptosis and reduced tumorigenicity of K562cells in vivo.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2002年第1期11-14,共4页
Acta Academiae Medicinae Sinicae
