Expression of socs1 and shp1 in JAK2V617F mutation positive myeloproliferative neoplasms and regulation effect of ruxolitinib / 第二军医大学学报

ABSTRACT

Objective To analyze the relationship between the JAK2V617F mutation and the expressions of phosphorylated Janus kinase 2 (p-JAK2), suppressor of cytokine signaling 1 (SOCS1), and SH2-containing protein tyrosine phosphatase 1 (SHP1) in JAK2V617F mutation positive myeloproliferative neoplasm (MPN) tissues, and to investigate the effects of JAK2 inhibitor ruxolitinib on regulating the proliferation of JAK2V617F mutation positive human erythroleukemia cell lines HEL and the expressions of SOCS1 and SHP1 in HEL cells. Methods A total of 48 patients with JAK2V617F mutation positive MPN (MPN group) and 24 patients with anemia (control group) in Hebei General Hospital and The First People’s Hospital of Baoding from Jul. 2012 to Aug. 2016 were enrolled in this study. The protein expressions of p-JAK2, SOCS1 and SHP1 in bone marrow biopsies (BMBs) were detected by immunohistochemistry. The correlations between JAK2V617F mutation level and the protein expressions of SOCS1 and SHP1 were analyzed by Spearman rank correlation analysis. HEL cells were treated with ruxolitinib at different concentrations (50, 100, 250, 500 and 1 000 nmol/L), and the viability of cells was determined by CCK-8 assay. The JAK2V617F mutation levels in MPN tissues and HEL cells and the mRNA expressions of JAK2, SOCS1 and SHP1 in HEL cells were detected by qPCR. The protein expressions of JAK2, SOCS1 and SHP1 in HEL cells were detected by Western blotting analysis. Results The ratio of JAK2V617F/JAK2 was (57.33±20.82)% in the MPN group and was zero in the control group. The protein expressions of p-JAK2, SOCS1 and SHP1 in BMBs of MPN patients were significantly different from those in the control group (all P0.01). The protein expressions of SOCS1 and SHP1 were negatively correlated with the mutation level of JAK2V617F (r＝－0.648, －0.692; P0.05). The expressions of SOCS1 and SHP1 in MPN patients with JAK2V617F/JAK250% were significantly higher than those in MPN patients with JAK2V617F/JAK2≥50% (P0.01), while the expression of p-JAK2 was significantly lower than that in MPN patients with JAK2V617F/JAK2≥50% (P0.01). After treatment with 250 nmol/L ruxolitinib for 24 h, 48 h, and 72 h, the viabilities of HEL cells were (60.06±3.87)%, (52.05±2.88)%, and (36.43±2.01)%, respectively. With the increase of ruxolitinib concentrations, the mRNA and protein expressions of JAK2 and the protein expression of p-JAK2 were gradually decreased (P0.01, P0.05), while the mRNA and protein expressions of SOCS1 and SHP1 were gradually increased (all P0.01). Conclusion Ruxolitinib can inhibit the expressions of JAK and the phosphorylation of JAK in HEL cells, enhance the expressions of SOCS1 and SHP1, and reduce the viability of HEL cells.