Oe Tomoya, Nagashima Takeyuki, Muramoto Masakazu, Yamazaki Takao, Morikawa Noriyuki, Okitsu Osamu, Nishimura Shintaro, Aoki Toshiaki, Katayama Yoshiki, Kita Yasuhiro
Pharmacology Research Laboratories, Astellas Pharma Inc., 21 Miyukigaoka, Tsukuba, Ibaraki 305-8585, Japan.
Neuropharmacology. 2006 Jun;50(8):1041-7. doi: 10.1016/j.neuropharm.2006.02.004. Epub 2006 Mar 30.
Neurite outgrowth plays a key role in neuronal development and regeneration, and is the hallmark assay for the effects of neurotrophic factors such as nerve growth factor (NGF). However, measuring neurite outgrowth is a slow and resource-intensive process. We therefore wanted to identify surrogate biomarkers for neurite outgrowth activity by gene expression analysis in SH-O10 cells, a subclone of the human SH-SY5Y neuroblastoma cell line but with much higher NGF-induced neurite outgrowth activity. Microarray analysis identified seven genes where mRNA levels were changed. NGF-induced decreases in levels of two genes, CyclinB2 and BIRC5, were confirmed by quantitative real-time RT-PCR. Levels of NGF-induced decreases in CyclinB2 and BIRC5 mRNA in several SH-SY5Y subclones with different neurite outgrowth responses correlated with their neurite outgrowth activities. Decreases in CyclinB2 and BIRC5 mRNA induced by FK506 or retinoic acid, both of which exert potentiation of NGF-induced neurite outgrowth effects but with different mechanisms, also correlated with their neurite outgrowth activities. In conclusion, decreasing levels of CyclinB2 and BIRC5 mRNA strongly correlate with neurite outgrowth activities in terms of NGF-related effect in SH-SY5Y subclonal cells, and have potential to become quantitative surrogate biomarkers for measuring NGF-related neurite outgrowth.
神经突生长在神经元发育和再生中起关键作用，并且是诸如神经生长因子（NGF）等神经营养因子作用的标志性检测方法。然而，测量神经突生长是一个缓慢且资源密集的过程。因此，我们希望通过对SH-O10细胞进行基因表达分析来鉴定神经突生长活性的替代生物标志物，SH-O10细胞是人类SH-SY5Y神经母细胞瘤细胞系的一个亚克隆，但具有更高的NGF诱导的神经突生长活性。微阵列分析鉴定出七个基因的mRNA水平发生了变化。通过定量实时RT-PCR证实了NGF诱导的两个基因CyclinB2和BIRC5水平的降低。在几个具有不同神经突生长反应的SH-SY5Y亚克隆中，NGF诱导的CyclinB2和BIRC5 mRNA水平降低与其神经突生长活性相关。FK506或视黄酸诱导的CyclinB2和BIRC5 mRNA降低，这两者均增强了NGF诱导的神经突生长效应，但机制不同，也与其神经突生长活性相关。总之，就SH-SY5Y亚克隆细胞中与NGF相关的效应而言，CyclinB2和BIRC5 mRNA水平的降低与神经突生长活性密切相关，并且有可能成为测量与NGF相关的神经突生长的定量替代生物标志物。
