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Objective To assess the effects of both TrkA and p75NTR on nerve growth factor (NGF)-induced differentiation of neuroblastoma cells.Methods Retroviral vectors were constructed to express the high affinity NGF receptor (TrkA) and low affinity NGF receptor (p75NTR). Neuroblastoma cell line IMR-32 was transfected by the vectors expressing either TrkA or p75NTR or both by using lipofectmine?reagent separately or cotransfected at the same time. Southern blot, Northern blot, RT-PCR and flow cytometry were used to determine the success of the transfection. MTT technique was to monitor the cell proliferation. Colony formation in soft agar and tumor forming assay in nude mice were used to test the biological characteristics of the tumor cells. Terminal-deoxynucleotidytransferase-mediated dUTP-biotin nick end labeling (TUNEL) assay was used to test the apoptosis of the tumor cells.Results Stable transformant cell lines expressing TrkA, p75NTR or both genes were established. Studies on these transformant cell line
Objective To assess the effects of both TrkA and p75NTR on nerve growth factor (NGF) -induced differentiation of neuroblastoma cells. Methods Retroviral vectors were constructed to express the high affinity NGF receptor (TrkA) and low affinity NGF receptor (p75NTR). Neuroblastoma cell line IMR-32 was transfected by the vectors containing either TrkA or p75NTR or both by using lipofectmine ™ reagent separately or cotransfected at the same time. Southern blot, Northern blot, RT-PCR and flow cytometry were used to determine the success of the transfection . MTT technique was to monitor the cell proliferation. Colony formation in soft agar and tumor forming assay in nude mice were used to test the biological characteristics of the tumor cells. Terminal-deoxynucleotidytransferase-mediated dUTP-biotin nick end labeling (TUNEL) assay was used to test the apoptosis of the tumor cells. Results Stable transformant cell lines expressing TrkA, p75NTR or both genes were established. Studies on these tran sformant cell line