Reverse Transcription PCR Screening of different neuronal guiding cues and their receptors in human staurosporine differentiated SH-SY5Y cells

摘要

Crucial roles of netrins and semaphorins and their receptors in neuronal development have been reported. For further research without using animal models it is important to know if well established neuronal cell lines express such guiding cues and their receptors. Here we focused on netrin-1, netrin Gl, G2, semaphorin 3A, 3F, 7A, and their receptors, respectively. We screened the neuroblastoma cell line SH-SY5Y before and after differentiation with 20 nM staurosporine via Reverse Transcription Polymerase Chain Reaction and qualitative analysis by agarose gel electrophoresis. Additionally, we determined the breast cancer cell line T47D for identifying potential neuronal markers and representing other functions of the selected neuronal guiding cues outside the nervous system in an in-vitro model. Our results showed no expression of netrin-1 and its receptor DCC in SH-SY5Y cells. Interestingly netrin-1 gene expression could be detected in the breast cancer cells T47Ds. The netrin-1 receptors Unc5A, B and C were expressed in both, treated and untreated SH-SY5Y cells, whereas UncSD was only expressed in treated cells. Semaphorin 3A and 3F as well as the receptors neuropilin-2, plexinA2, A3 and plexinCl could be detected in differentiated and undifferentiated SH-SY5Y cells. Neuropilin-1 was only synthesized in differentiated neuroblastoma cells in contrast to plexinAl, which was present in undifferentiated SH-SY5Y. Therefore, Unc5C, neuropilin-1, and plexinCl could be identified as potential neuronal markers.