hes1-protein--human and Leukemia--Myelogenous--Chronic--BCR-ABL-Positive

Topics: Animals; Basic Helix-Loop-Helix Transcription Factors; Blast Crisis; Genes, abl; Hematopoietic Stem Cells; Homeodomain Proteins; Humans; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Mice; Molecular Targeted Therapy; Transcription Factor HES-1

To explore the relationship between the expression levels of JARID1B,Hes1 and MMP-9 genes and the stages of chronic myelogenous leukemia(CML) and the curative effect of imatinib mesylate (IM).. Peripheral blood samples of 15 cases of CML in chronic phase and 10 cases of CML in progressive phase were collected from the Hematology Department of Taihe Hospital affiliated to Hubei University of Medicine and 15 cases of healthy people in the Physical Examination Center. CML patients were divided into effective group and ineffective group based on the efficacy after treatment with IM, then real-time PCR was used to detect the expression levels of JARID1B, Hes1 and MMP-9 mRNA, finally, the differences in the level of gene expression and their correlations with CML stages and IM curative efficacy were analysed.. The expression levels of Hes1 and MMP-9 in initially diagnosed patients in chronic and progressive phase without IM treatment were significantly higher than those of health people（P＜0.05）. There was no significant difference in the expression level of JARID1B between chronic phase patients and health people（P＞0.05）, but the expression level of JARID1B in the progressive phase patients was higher than that of health people （P＜0.05）. The expression levels of JARID1B and Hes1 in the IM-effective group were not significantly different from those in the IM-ineffective group (P=0.85,P=0.82）, while the expression level of MMP-9 in the IM-effective group [JP2]was significantly lower than that in the IM-ineffective group（P＜0.05）.. The expression levels of JARID1B Hes1 and MMP-9 relate with the different phase of CML; The expression levels of JARID1B and Hes1 have not significant relationship with IM curative efficacy, the MMP-9 gene expression level relates with IM curative efficacy.. 伊马替尼治疗的慢性髓系白血病患者JARID1B、Hes1和MMP-9基因表达水平差异的研究.. 探讨JARID1B、Hes1和MMP-9基因表达水平与慢性髓系白血病（chronic myelogenous leukemia，CML）疾病的病程阶段及伊马替尼（imatinib mesylate, IM）疗效的关系.. 收集湖北医药学院附属太和医院血液科CML患者25例 (慢性期15例、进展期10例)和体检中心正常人15例的外周血标本。按初次确诊为CML患者的病期和服用IM后的疗效分组（有效组和无效组），采用实时定量RT-PCR方法检测JARID1B、Hes1和MMP-9基因mRNA表达水平，分析JARID1B、Hes1和MMP-9基因表达水平差异与疾病阶段及IM疗效的相关性。.. 初次确诊未服用IM慢性期和进展期CML患者Hes1和MMP-9基因的表达水平明显高于正常人（P＜0.05）；CML患者慢性期JARID1B基因的表达水平与正常人相比无明显差异（P=0.20），进展期患者JARID1B的表达水平明显高于正常人（P＜0.05）；IM治疗有效组JARID1B和Hes1基因的表达水平与无效组相比无明显差异（P=0.85、P=0.82），MMP-9的表达水平有效组明显低于无效组（P＜0.05）.. JARID1B、Hes1和MMP-9基因的表达水平与CML的不同病程分期有关；JARID1B、Hes1基因表达水平与IM疗效无关，MMP-9基因表达水平与IM疗效有关.

Topics: Antineoplastic Agents; Humans; Imatinib Mesylate; Jumonji Domain-Containing Histone Demethylases; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Matrix Metalloproteinase 9; Nuclear Proteins; Repressor Proteins; Transcription Factor HES-1

Tyrosine kinase inhibitor (TKI)-based therapy has created promising results among much chronic myeloid leukemia (CML) patients. Imatinib as a relatively specific inhibitor of Bcr-Abl is at present one of the undisputed therapeutic agent for newlydiagnosed patients with CML. However, the occurrence of imatinib-resistance enlightens the urgent need to identify other therapeutic agents against CML. Juglone (5-hydroxy-2-methyl-1, 4-naphthoquinone) exerts cytotoxic effects against various human cancer cell lines. However, the mechanisms through which Juglone induces anticancer effects in CML especially in comparison with imatinib treatment remain unknown. Our results revealed that Juglone-inhibited K562 cells growth through inducing apoptosis. Based on our Western blot analyses, Juglone significantly reduced p-Akt levels and increased the expression level of Forkhead box O1 (FoxO1) and FoxO3a proteins. Moreover, hairy/enhancer of split-1 (Hes1) protein, overexpressed under the influence of Juglone, is apparently involved in Juglone-induced apoptosis among K562 cells. Conversely, treatment with imatinib attenuated Hes1 protein expression. Considering the different functional mechanism of Juglone compared with imatinib, it seems that Juglone treatment could be a useful alternative strategy for the treatment of patients with imatinib-resistance.

Topics: Antineoplastic Agents; Apoptosis; Cell Proliferation; Cell Survival; Forkhead Box Protein O1; Forkhead Box Protein O3; G1 Phase Cell Cycle Checkpoints; Gene Expression Regulation, Leukemic; Humans; Imatinib Mesylate; K562 Cells; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Naphthoquinones; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-akt; Transcription Factor HES-1

The deregulation of lineage control programs is often associated with the progression of haematological malignancies. The molecular regulators of lineage choices in the context of tyrosine kinase inhibitor (TKI) resistance remain poorly understood in chronic myeloid leukemia (CML). To find a potential molecular regulator contributing to lineage distribution and TKI resistance, we undertook an RNA-sequencing approach for identifying microRNAs (miRNAs). Following an unbiased screen, elevated miRNA182-5p levels were detected in Bcr-Abl-inhibited K562 cells (CML blast crisis cell line) and in a panel of CML patients. Earlier, miRNA182-5p upregulation was reported in several solid tumours and haematological malignancies. We undertook a strategy involving transient modulation and CRISPR/Cas9 (clustered regularly interspersed short palindromic repeats)-mediated knockout of the MIR182 locus in CML cells. The lineage contribution was assessed by methylcellulose colony formation assay. The transient modulation of miRNA182-5p revealed a biased phenotype. Strikingly, Δ182 cells (homozygous deletion of MIR182 locus) produced a marked shift in lineage distribution. The phenotype was rescued by ectopic expression of miRNA182-5p in Δ182 cells. A bioinformatic analysis and Hes1 modulation data suggested that Hes1 could be a putative target of miRNA182-5p. A reciprocal relationship between miRNA182-5p and Hes1 was seen in the context of TK inhibition. In conclusion, we reveal a key role for miRNA182-5p in restricting the myeloid development of leukemic cells. We propose that the Δ182 cell line will be valuable in designing experiments for next-generation pharmacological interventions.

Topics: Cell Lineage; Cell Proliferation; Drug Resistance, Neoplasm; Erythroid Cells; Fusion Proteins, bcr-abl; Gene Expression Regulation, Leukemic; Humans; K562 Cells; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; MicroRNAs; Myeloid Cells; Protein Kinase Inhibitors; Transcription Factor HES-1

Notch signalling is critical for haemopoietic stem cell (HSC) self-renewal and survival. The role of Notch signalling has been reported recently in chronic myeloid leukaemia (CML) - a stem cell disease characterized by BCR-ABL tyrosine kinase activation. Therefore, we studied the relationship between BCR-ABL and Notch signalling and assessed the expression patterns of Notch and its downstream target Hes1 in CD34+ stem and progenitor cells from chronic-phase CML patients and bone marrow (BM) from normal subjects (NBM). We found significant upregulation (p<0.05) of Notch1, Notch2 and Hes1 on the most primitive CD34+Thy+ subset of CML CD34+ cells suggesting that active Notch signalling in CML primitive progenitors. In addition, Notch1 was also expressed in distinct lymphoid and myeloid progenitors within the CD34+ population of primary CML cells. To further delineate the possible role and interactions of Notch with BCR-ABL in CD34+ primary cells from chronic-phase CML, we used P-crkl detection as a surrogate assay of BCR-ABL tyrosine kinase activity. Our data revealed that Imatinib (IM) induced BCR-ABL inhibition results in significant (p<0.05) upregulation of Notch activity, assessed by Hes1 expression. Similarly, inhibition of Notch leads to hyperactivation of BCR-ABL. This antagonistic relationship between Notch and BCR-ABL signalling was confirmed in K562 and ALL-SIL cell lines. In K562, we further validated this antagonistic relationship by inhibiting histone deacetylase (HDAC) - an effector pathway of Hes1, using valproic acid (VPA) - a HDAC inhibitor. Finally, we also confirmed the potential antagonism between Notch and BCR/ABL in In Vivo, using publically available GSE-database, by analysing gene expression profile of paired samples from chronic-phase CML patients pre- and post-Imatinib therapy. Thus, we have demonstrated an antagonistic relationship between Notch and BCR-ABL in CML. A combined inhibition of Notch and BCR-ABL may therefore provide superior clinical response over tyrosine-kinase inhibitor monotherapy by targeting both quiescent leukaemic stem cells and differentiated leukaemic cells and hence must be explored.

Topics: Antigens, CD34; Apoptosis; Basic Helix-Loop-Helix Transcription Factors; Blotting, Western; Cell Cycle; Cell Proliferation; Fusion Proteins, bcr-abl; Homeodomain Proteins; Humans; Imatinib Mesylate; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Neoplastic Stem Cells; Protein Kinase Inhibitors; Real-Time Polymerase Chain Reaction; Receptor, Notch1; Receptor, Notch2; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transcription Factor HES-1; Tumor Cells, Cultured

High levels of HES1 expression are frequently found in BCR-ABL(+) chronic myelogenous leukemia in blast crisis (CML-BC). In mouse bone marrow transplantation (BMT) models, co-expression of BCR-ABL and Hes1 induces CML-BC-like disease; however, the underlying mechanism remained elusive. Here, based on gene expression analysis, we show that MMP-9 is upregulated by Hes1 in common myeloid progenitors (CMPs). Analysis of promoter activity demonstrated that Hes1 upregulated MMP-9 by activating NF-κB. Analysis of 20 samples from CML-BC patients showed that MMP-9 was highly expressed in three, with two exhibiting high levels of HES1 expression. Interestingly, MMP-9 deficiency impaired the cobblestone area-forming ability of CMPs expressing BCR-ABL and Hes1 that were in conjunction with a stromal cell layer. In addition, CMPs expressing BCR-ABL and Hes1 secreted MMP-9, promoting the release of soluble Kit-ligand (sKitL) from stromal cells, thereby enhancing proliferation of the leukemic cells. In accordance, mice transplanted with CMPs expressing BCR-ABL and Hes1 exhibited high levels of sKitL as well as MMP-9 in the serum. Importantly, MMP-9 deficiency impaired the development of CML-BC-like disease induced by BCR-ABL and Hes1 in mouse BMT models. The present results suggest that Hes1 promotes the development of CML-BC, partly through MMP-9 upregulation in leukemic cells.

Topics: Animals; Basic Helix-Loop-Helix Transcription Factors; Blast Crisis; Bone Marrow Transplantation; Cell Movement; Cell Proliferation; Flow Cytometry; Fusion Proteins, bcr-abl; Gene Expression Regulation, Leukemic; Homeodomain Proteins; Humans; Kaplan-Meier Estimate; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Matrix Metalloproteinase 9; Mice; Mice, 129 Strain; Mice, Inbred C57BL; Mice, Knockout; Models, Genetic; NF-kappa B; Oligonucleotide Array Sequence Analysis; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Transcription Factor HES-1; Up-Regulation

Hairy enhancer of split 1 (Hes1) is a basic helix-loop-helix transcriptional repressor that affects differentiation and often helps maintain cells in an immature state in various tissues. Here we show that retroviral expression of Hes1 immortalizes common myeloid progenitors (CMPs) and granulocyte-macrophage progenitors (GMPs) in the presence of interleukin-3, conferring permanent replating capability on these cells. Whereas these cells did not develop myeloproliferative neoplasms when intravenously administered to irradiated mice, the combination of Hes1 and BCR-ABL in CMPs and GMPs caused acute leukemia resembling blast crisis of chronic myelogenous leukemia (CML), resulting in rapid death of the recipient mice. On the other hand, BCR-ABL alone caused CML-like disease when expressed in c-Kit-positive, Sca-1-positive, and lineage-negative hematopoietic stem cells (KSLs), but not committed progenitors CMPs or GMPs, as previously reported. Leukemic cells derived from Hes1 and BCR-ABL-expressing CMPs and GMPs were more immature than those derived from BCR-ABL-expressing KSLs. Intriguingly, Hes1 was highly expressed in 8 of 20 patients with CML in blast crisis, but not in the chronic phase, and dominant negative Hes1 retarded the growth of some CML cell lines expressing Hes1. These results suggest that Hes1 is a key molecule in blast crisis transition in CML.

Topics: Animals; Basic Helix-Loop-Helix Transcription Factors; Blast Crisis; Cell Line, Transformed; Fusion Proteins, bcr-abl; Granulocyte-Macrophage Progenitor Cells; Homeodomain Proteins; Interleukin-3; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Mice; Neoplastic Stem Cells; Rats; Repressor Proteins; Transcription Factor HES-1