Role of NF-?B in HGF-mediated proliferation of WB F-344 cell / 解放军医学杂志

ABSTRACT

Objective To study the role of NF-?B pathway in HGF (hepatocyte growth factor)-mediated proliferation of WB F-344 cell. Methods WB F-344 cells were treated with HGF in different concentrations i.e. 0, 20, 40, 80 and 100ng/ml. After incubation for 24 hours, proliferation effect was analyzed with [3H] Thymidine. WB F-344 cells transfected with I?B? plasmid mutants by liposome transfection were used to block NF-?B activity, and then they were treated with 40ng/ml HGF for 0, 15, 30, 60 and 120min, respectively. The nucleoprotein was extracted to determine the NF-?B DNA-binding activity by EMSA. WB F-344 cells were also transiently transfected with BD Great EscA PeTM SEAP vector with liposome and then stimulated with HGF in different concentrations (0, 10, 20, 40ng/ml) for 8 hours to detect the NF-?B transcription activity. WB F-344 cells were pre-treated with NF-?B inhibitor BAY-11-7082 or ASA, and then with 40 ng/ml HGF for 24 hours to analyze the proliferation effect by [3H] Thymidine. Results HGF promoted the proliferation of WB F-344 cells and exhibited positive dose-effect tendency. HGF could enhance the NF-?B DNA-binding activity. NF-?B complexes appeared 15min after addition of HGF. The complexes reached the peak value 30-60min after addition of HGF and then decreased gradually. HGF could also enhance the NF-?B transcription activity, exhibiting positive dose-effect tendency. HGF-mediated proliferation was remarkably inhibited when NF-?B activity was blocked by BAY-11-7082 or ASA. In WB F-344 cells transfected with p-CMV-I?B?, the proliferative response to HGF was completely interrupted. Conclusions HGF may promote the proliferation of WB F-344 cells in a positive dose-effect tendency. The activation of NF-?B is necessary for HGF-mediated proliferation of WB F-344 cells.