Effects of Ruxolitinib on Myeloproliferative Neoplasms via the Negative Regulators.

BACKGROUND: We evaluated the JAK2V617F mutation and p-JAK2, SOCS-1, SHP-1 expression in JAK2V617F positive myeloproliferative neoplasms (MPNs) patients and the role of JAK/STAT pathway in human erythroleukemia (HEL) cells, which had JAK2V617F mutation. METHODS: Protein expression of p-JAK2, SOCS-1, SHP-1 in bone marrow biopsies (BMBs) were detected by immunohistochemical staining methods. Cell apoptosis and cell cycle were detected by flow cytometry and Caspase 3/7 assay kits. RESULTS: 1. The p-JAK2, SOCS-1, and SHP-1 expressions were significantly different between JAK2V617F positive MPN and control patients (p < 0.01); 2. After being treated for 3 months, the p-JAK2, SOCS-1, and SHP-1 expressions were significantly different compared with newly diagnosed patients (p < 0.01). 3. HEL cell viabilities were significantly different after being treated with different concentrations of ruxolitinib. Ruxolitinib had a significant effect on the cell apoptosis, viability, and the protein activity of caspase-3 and -7 of HEL cells. 3. The mRNA and protein expressions of JAK2 and the protein expression of p-JAK2 were gradually decreased (p ＜ 0.01, p ＜ 0.05), while the mRNA and protein expressions of SOCS1 and SHP1 were gradually increased (all p ＜ 0.01).

[Effect of IFN-α2b on COX-2 and Angiogenesis in JAK2V617F Mutation Myeloproliferative Neoplasms].

OBJECTIVES: To investigate the influence of interferon-alpha-2b (IFN-α2b) with JAK2 kinase, COX-2 and microvessel density in patients of MPN and the relation of JAK2V617F and COX-2 in human erythroleukemia cell line (HEL) cells. METHODS: Forty-two cases of MPN patients with JAK2V617F mutation of initial treatment were collected from the Frist hospital of Baoding, including the IFN-α2b treatment group with 17 cases and untreated group with 25 cases. 10 cases of idiopathic immune thrombocytopenic purpura (ITP) patients synchronization were enrolled as controls. JAK2V617F/JAK2 mutation burden of MPN patients was detected by real time PCR (qRT-PCR);the expression levels of p-JAK2, COX-2 and microvascular density (MVD) marked with CD105 inpathological tissues of bone marrow in patients of MPN and ITP were detected by immunohistochemistry. The HEL cells were treated with different concentrations of IFN-α2b. The cell proliferation inhibition rate was calculated by CCK-8 test;the apoptosis rate was detected by flow cytometry; cell migration ability was tested by transwell chambers. JAK2 and COX-2 mRNA were detected by semi-quantitative PCR; p-JAK2 and COX-2 protein in HEL cells were detected by Western blotting. RESULTS: The expression levels of p-JAK2, COX-2 protein and MVD in untreated group were significantly higher than those of control groups. p-JAK2, COX-2 and MVD levels were significantly reduced in patients treated with IFN-α2b. Cell growth inhibition rates and apoptosis rates raise up by dose of IFN-α2b in HEL cells at 48 h.The mRNA expression levels of JAK2 and COX-2 as well as protein expression levels of p-JAK2 and COX-2 had a decreasing tendency with the increase of IFN-α2b concentration at 48 h.The migration capacity level of HEL cells which treated with 0.5×10 4 U/L IFN-α2b after 24 h was lower than that of control group. CONCLUSIONS: Angiogenesis of MPN and COX-2 were inhibited by IFN-α2b which regulates JAK2 signal pathway.

[Clinical analysis of acute myeloid leukemia with t(8;21) (q22;q22) and loss of Y chromosome].

This study was aimed to investigate the clinical characteristics of acute myeloid leukemia (AML) with t (8;21) (q22;q22) and loss of Y chromosomes. Clinical data of 267 cases of AML were collected from January 2010 to June 2013. Among 267 AML, there were 13 cases with t (8;21) (q22;q22) and loss of Y chromosomes. The clinical data including clinical indicators, treatment protocols, curative effect and prognosis were analyzed retrospectively. The results showed that after normalized chemotherapy, there were 4 patients with complete remission at the first cycle of treatment, 4 patients with complete remission at the second cycle, 4 patients with complete remission at the third cycle, but one patient without complete remission after 4 cycles. There were 6 patients who did not relapse during consolidation and intensive therapy. Among these 6 patients, 4 cases accepted chemotherapy combined with transplantation, other 2 cases accepted chemotherapy. In the remainder 6 patients, 4 cases relapsed once, one cases relapsed twice, 1 cases relapsed for three times. Moreover, 2 cases who accepted the chemotherapy and auto-hematopoietic stem cell trans-plantation, were diagnosed as relapse, after accepted allo-hematopoietic stem cell transplantation, currently are in disease-free status. In follow-up period, the relapse-free survival (RFS) time was 4.67 ± 3.45 months in chemotherapy group, the RFS time is 34.17 ± 21.37 months in chemotherapy and transplantation group. The chemotherapy combined with transplantation extended the RFS time (P < 0.05). It is concluded that the NCCN guide indicates that AML with t (8;21) ( q22;q22) showed a good prognosis. but the clinical course of treatment confirmed that the prognosis of AML patients with t (8;21) (q22;q22) and loss Y chromosomes is poor, including uneasy remission and easy relapse, for improving the prognosis of these patients, the hematopoietic stem cell transplantation should be recommended.