神经生长因子诱导神经母细胞瘤分化的研究

目的观察神经生长因子（NGF）对神经母细胞瘤（neuroblastoma，NB）细胞增殖的抑制和可能的分子机制及NGF-TrkA信号传导途径在NB细胞分化中的作用。方法取本院住院患儿新鲜手术标本，进行原代细胞培养作为细胞模型，通过台盼蓝拒染法计数活细胞；观察NGF干预前后细胞形态学变化；MTT法检测细胞的增值活性；RT-PCR分析TrkA表达情况。结果NGF作用NB细胞后，细胞形态发生显著变化，并可剂量依赖性的抑制细胞增殖，2d时TrkA表达增加，4d以后出现TrkA表达的明显增加。结论NGF对NB细胞体外增殖有抑制作用并诱导其分化，且表现剂量依赖关系；NGF可诱导NB细胞分化成熟及TrkAmRNA表达水平增高，可能是NGF诱导NB细胞分化逆转的分子生物学机制之一。

Study on differentiation and proliferation of neuroblastoma(NB) cells induced by nerve growth factor(NGF)

Objective To investigate the differentiation and proliferation of neuroblastoma（NB） cells induced by nerve growth factor（NGF） and its possible mechanisms. Methods NB tumor tissues were collected and primary cell culture was performed. Trypan blue exclusion was used to count the alive cells. Morphological changes were observed under the phase - contrast microscope. The change of cell proliferation was determined by means of MTT assay. TrkA expression was examined by RT - PCR method. Results By day 2, the alive cells of group A, B and C decreased by treatment of NGF; morphological changes of NB cells were also observed. RT - PCR detection showed that the TrkAmRNA expression increased from day 2 and reached its peak on day 8 in a time and dose dependent mariner by treatment of NGF. Conclusion NB cells treated by NGF differentiate to neuron - like cells and the cell growth is inhibited in a dose- dependent mariner. TrkA＇s expression is increased by NGF. This may be one of the mechanisms of spontaneous regression of NB.