全反式维甲酸体外诱导神经母细胞瘤细胞分化研究

Experimental Study of Induce Differentiation with All Trans Retinoic Acid in Human Neuroblastoma Cells

[目的]观察全反式维甲酸(alltransretinoicacid,ATRA)体外对人神经母细胞瘤细胞系SK N SH细胞生长的影响并对其作用机制进行初步探讨。[方法]SK N SH细胞经ATRA处理后 ,采用相差显微镜、神经纤维银染法、透射电镜观察、逆转录PCRmRNA检测等手段 ,观察ATRA对SK N SH细胞生长的影响。[结果]ATRA作用SK N SH细胞12天后 ,细胞形态发生显著变化 ,表现为明显分化 ;出现神经原性表型 ,多个细胞形成神经节 ,节间形成粗而长的类神经纤维 ;尼氏小体和银染法亦证明SK N SH细胞产生显著分化。透射电镜观察结果表明 ,ATRA对该细胞无明显凋亡诱导作用。ATRA对SK N SH细胞的维甲酸受体 β(RARβ)表达水平有上调作用。[结论]ATRA对神经母细胞瘤细胞体外能产生明显诱导分化作用 ,此作用与ATRA上调细胞的RARβ

To investigate the antitumor mechanism of ATRA in SK N SH cells.The human neuroblastoma cell line SK N SH and ATRA were used.Nissl body staining and Sevier Munger staining,TEM observation,RT PCR assay were carried out in this study.Differentiation morphological changes of SK N SH cells were observed by phase contrast microscopy and demonstrated by Nissl body staining and Sevier Munger staining after 12 days of treatment at the concentration of 1μM ATRA .By day 6,the cell bodies were very small with long axonal processes between one another.By day 12,the cells formed isolated neural crest cells,which resembled the differentiated neurons and extended axon like neurites.No significant cells apoptosis induced by ATRA.The results of this study also showed that the expression of RARβ gene in SK N SH cells was up regulated by ATRA .[Conclusion]Inhibiting the proliferation and inducing differentiation of human neuroblastoma cells(SK N SH)by ATRA may result from up regulating the expression level of RARβ mRNA.Our results may also support the hypothesis of use of the ATRA in the treatment of human neuroblastoma.