To Discuss The Effect Of PI3K/AKT Pathway On Cell Proliferation And Differentiation Of Human Neuroblastoma In Vitro

PART1The study of PI3K/Akt signal pathway in neuroblastomacell proliferationObjective: To investigate the role of the PI3K/Akt signal pathway inneuroblastoma cell proliferation.Methods: Human neuroblastoma cell lines, SK-N-SH were culturedin DMEM media supplemented with10%fetal bovine serum (FBS) in ahumidified atmosphere of5%CO2at37℃,(1-5)×105/ml cells wereseeded per well in a96-well plate,100ul per well. After24hours, variousdoses of LY294002were given to cells; other Cells were changed inserum-free medium for a further12h before treating various doses of IGF-1.then,after24h,48h LY294002were given, and12h,24h after IGF-1weregiven, the cell survival rate was measured by MTT.Results:24h and48h after LY294002treatment,a statisticallysignificant decrease of cell survival rate was observed between theexperimental group and control group. negative co-relationship was foundbetween the cell survival rate and the dose and time of LY294002.12h,24h after IGF-1treatment,a statistically significant increase of cell survival ratewas observed between the experimental group and control group.but noco-relationship was found between the cell survival rate and the time ofIGF-1.Conclusion: our results suggest that PI3K/Akt signaling pathway isclosely related to the proliferation of neuroblastoma cells. the inhibition ofthe PI3K/Akt can significantly inhibit the neuroblastoma cells proliferation,the PI3K/Akt signaling pathway agonists can promote the proliferation ofneuroblastoma cells. Part2: The study of PI3K/Akt signal pathway in neuroblastomacell differentiationObjective: To investigate the PI3K/Akt signal pathway inneuroblastoma cell differentiation.Methods: Human neuroblastoma cell lines, SK-N-SH were cultured inDMEM media supplemented with10%fetal bovine serum (FBS) in ahumidified atmosphere of5%CO2at37℃,(1-5)×106/ml cells wereseeded in a6-well plate,2ml per well. After24hours, LY294002wasgiven to cells, other Cells were changed in serum-free medium for a further 12h before treating IGF-1. then,48h after LY294002was given, and12hafter IGF-1was given, cell morphology changes were observed by invertedmicroscope, the expression of TrkA mRNA were assessed by fluorescentquatititive PCR（FQ-PCR）.Results:after LY294002and IGF-1treatment, no Morphologicalchange of neuroblastoma cell differentiation was observed．compared toDMSO control group and blank control group, fluorescent quatititive PCR（FQ-PCR） showed that the expressions of TrkA mRNA wasdown-regulated by LY294002，the difference was statistically significant. nostatistically significant difference was found between IGF-1group andcontrol group;Conclusion: PI3K/AKT signal the pathway may be involved in theprocess of differentiation of neuroblastoma cells, the inhibition of thePI3K/Akt signal pathway may inhibit neuroblastoma cell differentiation.