Investigation of the effects of rapamisin on mouse neuroblastoma (NB2a) cell culture

Aim: The aim of this study is to investigate the neurotoxic or neuroprotective effects of an active substance called Rapamycin, which is an mTOR inhibitor, in mouse neuroblastoma cell line (NB2A). Material and Methods: In present study, the possible neurotoxic effects of Rapamycin were investigated in mouse neuroblastoma cell (NB2A) array. Cells were cultivated in an incubator by adding DMEM in a flask. Rapamycin was dissolved in DMSO and diluted with medium before using the stock solution. A dose response curve was generated using the CellTiter-Glo® luminescence cell viability measurement method. On day 1, NB2A cells were added to all wells and incubated for 24 hours. After 24 hours, the medium was withdrawn from all the wells and except the first three wells and the differentiation medium was added. Except for the first three (+ control) wells containing differentiation medium, the others were loaded with Rapamycin at different concentrations. Cells were incubated for 1 day. Then the wells were stained with Coomassie Blue and measurements were made using an image analysis program. In order to investigate the effects of Rapamycin on the cell proliferation and neurotoxicity, MTT and NTT tests were performed by adding chlorpyrifos at 25 µM concentrations. Statistical analyzes were performed and p<0.05 was chosen as the significance level. Findings: In the NTT test, neurite prolongation was not significant, and in the MTT test, significant cell proliferation reduction was observed. A dose-dependent decrease in NB2A cells was also observed with neurotoxic chlorpyrifos. Results: It has been shown that our active ingredient, rapamycin, is neurotoxic and can cause neuronal damage and death. However, since the cell culture we used is x composed of cancer cells, it was thought that it might have an anticarcinogenic effect. Keywords: Rapamycin, mouse, neuroblastoma, cell culture, mTOR, MTT, NTT, Chlorpyrifos, neurotoxicity, neuron, neurodegenerative diseases.