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Background: Neuroblastoma (NB) is a very common childhood malignant tumor with considerable morbidity and mortality. Nevertheless, some cases can disappear spontaneously and be induced to differentiate into mature cells in vitro. It may be a potential treatment method to induce NB cells to differentiate into mature cells. This study was undertaken to observe the differentiation of neuroblastoma cell line transfected with nerve growth factor (NGF) gene and investigate the role of NGF in the differentiation.
Methods: Tumor specimens of NB patients were collected for primary cell culture and the cells were separated and purified to be cell line as a cell model. The plasmid containing NGF gene was transfected into NB cells mediated by liposome. Morphological changes were observed under a phase-contrast microscope. Cell proliferation was determined by the MTT method and mitosis index.
Results: The expression of NGF was higher after gene transfection, the cell proliferation was inhibited and morphological changes occurred. Total RNA separated by agrose gel electrophoresis resulted in clear 28s and 18s, indicating that RNA quality was good. RT-PCR products about 726 bp, matching NGF size, indicated that NB cells can express a certain amount of NGF, and the expression was increased after NGF gene transfection. Forty-eight hours after NGF gene transfection, morphodifferentiation presented like neurodendrite and axons, which were similar to mature ganglion cells.
Conclusions: The established NB cell line is an induced type (type N). The transgenic NB cells can highly express NGF, which can inhibit proliferation and induce differentiation.
Key words: NGF; transfection; neuroblastoma; cell line; differentiation
World J Pediatr 2007;3(2):115-120
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