Role of NF-κB in apoptosis of immortalized neural progenitor cells / 中华麻醉学杂志

ABSTRACT

Objective To investigate the role of NF-κB in apoptosis of immortalized neural progenitor cells (INPCs) . Methods INPCs were cultured in 6-well plates and were randomly divided into 5 groups ( n = 6 each) group I was not transfected with any plasmid (group INPC); group Ⅱ was transfected with control plasmid (group INPC/CMV); group Ⅲ was transfected with plasmid RcCMV-p50 (group INPC/p50); group Ⅳ was transfected with plasmid RcCMV-p65 (group INPC/p65) and group V was transfected with plasmid RcCMV-p50 and RcCMV-p65 (group INPC/p50p65). Group INPC/CMV ( H ), INPC/p50 (Ⅲ) and INPC/p65 (Ⅳ) were screened by G418, and the positive clones were then cultured for 3-4 weeks. The transcription of p50 mRNA or p6S mRNA was detected by RT-PCR. The NF-κB activity was measured by luciferase reporter gene assay. The cell apoptosis was measured by annexin V/PI staining. In group INPC/p50p65 and group INPC/p65, the cultured positive clone was transiently transfected with plasmid RcCMV-p50. Two days after transfection, the same measurement was performed in group INPC/pS0p65 and the other groups. Results The expression of p50 mRNA was significantly increased in group INPC/p50 and INPC/p50p65 as compared with the other groups ( P < 0.05) . The expression of p65 mRNA, the NF-κB activity and the apoptotic rate were significantly increased in group INPC/p65 and INPC/p50p65 as compared with the other groups ( P < 0.05). Conclusion Enhanced NF-κB activity can increase immortalized neural progenitor cell apoptosis.