高亲和性神经生长因子受体对神经生长因子诱导神经母细胞瘤细胞分化的影响  

Effects of high-affinity nerve growth factor (NGF) receptor gene on NGF induced differentiation of neuroblastoma cell line

作　　者：折晓宁[1] 陈杰[1] 刘彤华[1] 高洁[1] 

机构地区：[1]中国医学科学院中国协和医科大学北京协和医院病理科,北京100730

出　　处：《中华病理学杂志》2000年第3期196-199,共4页Chinese Journal of Pathology

基　　金：国家杰出青年基金资助项目!(39625012);国家教委跨世纪优秀人才计划基金资助项目

摘　　要：目的　观察人神经母细胞瘤细胞系IMR 32细胞在恢复高亲和性神经生长因子受体基因trkA表达后对神经生长因子 (NGF)诱导分化的反应。方法　应用基因重组技术构建含外源trkAcDNA的逆转录病毒载体 ,经PA31 7细胞包装后感染靶细胞系IMR 32细胞 ,经Southernblot杂交、逆转录 聚合酶链反应 (RT PCR)证实转化细胞系中含有外源基因的整合及稳定表达后 ,进行神经生长因子诱导分化实验。结果　转化细胞系未经NGF诱导前与母系细胞在细胞形态及生长状态方面差异均无显著性意义 ,而经NGF诱导后细胞出现明显神经元样分化 ,生长及代谢速率减慢 (四甲基偶氮唑盐法测定原细胞系IMR 32和空载体转化细胞系IMR 32 /pIRV分别为 0 .2 5 8± 0 .0 1 7,0 .2 37± 0 .0 1 1 ;而trkA基因转化的细胞系则仅为 0 .0 2 8± 0 .0 0 3) ,撤去NGF后分化细胞仍然维持分化状态。转化细胞在软琼脂内很少形成集落 ,集落形成率仅为 0 .6 ;远低于原细胞系IMR 32和空载体转化细胞系IMR 32 /pIRV的 32 .3和 33.6 ,在裸鼠体内未见肿瘤形成。结论　高亲和性神经生长因子受体基因的表达可以使转化细胞系对NGF产生不可逆的诱导分化反应 。Objective To study the effects of transfection of high affinity nerve growth factor receptor gene (trkA) on NGF induced differentiation of human neuroblastoma cell line IMR 32. Methods The recombinant retrovirus vector containing exogeneous trkA gene was constructed and packed by PA317 packaging cell line. The neuroblastoma cell line IMR 32 was transfected by virus containing supernatant. The transformant cell line was confirmed by Southern blot and RT PCR techniques. The NGF was used to induce cellular differentiation of the transformant cells. Results The trkA gene was successfully transferred and expressed in the neuroblastoma cells. After NGF treatment, the transformant cells displayed apparent neuron like differentiation morphologically, and a slower rate of cell growth (MTT value 0.028±0.003) compared with original cell line (0.258±0.017) and empty virus transformed cell line (0.237±0.011). The cells remained in differentiated status after withdrawing the NGF from the medium. The transformant tumor cells rarely formed colonies in soft agar and failed to form tumor in nude mice. Conclusion Restoration of high affinity NGF receptor (trkA) expression in neuroblastoma cells could induce non reversal differentiation. The trkA might be the important factor during NGF induced differentiation of neuroblastoma cell.

关 键 词：神经母细胞 神经生长因子受体 基因转移 分化 

分 类 号：R739.4[医药卫生—肿瘤]