Regulatory effect of dissociative intracytoplasmic domain of leukemia inhibitory factor receptor ? subunit on growth of HL-60 cells / 第二军医大学学报

ABSTRACT

Objective:To investigate the regulatory effect of dissociative intracytoplasmic domain of leukemia inhibitory factor receptor ? (LIFR?) subunit on the growth of HL-60 cells and the related signal transduction mechanism. Methods: The constructed recombinant plasmids pcDNA3.0-190CT2+3 and cDNA3.0-190CT3 were identified and transfected into HL-60 cells with liposome FuGene-6. Then the cells were cultured in a medium containing G-418 and the positive clones were selected. Immunohistochemical method and Western blotting were used to detect the expression of the target protein in HL-60 cells and the growth curve of the cells was plotted. The expression of proliferating cell nuclear antigen (PCNA) and the phosphorylation levels of signal transducer and activator of transcription 3 (Stat3) were assayed by Western blotting. Results: Western blotting detected the band of target protein and the cell line stably expressing the target protein was obtained. Compared with that of the wild type group, the cell sizes in 190CT2+3 and 190CT3 group were enlarged. The ratio of leaf cell number to the total cell number increased and the cell proliferation was slowed down. We also found that the expression of PCNA was decreased and the phosphorylation level of STAT3 was increased. Conclusion: The dissociative intracytoplasmic domain of LIFR? subunit can accelerate the differentiation of HL-60 cells, inhibit the proliferation of HL-60 cells, and activate signal molecule STAT3.