Journal of

Background and Aim : Myeloproliferative neoplasms are clonal and heterogeneous disorders of hematopoietic stem cells lead to increase of one or more cell lines in the blood. Recently, the acquired mutation JAK2 V617F has been described in the majority of patients with myeloproliferative neoplasms (MPNs).This mutation is characterized by a G to T transverse at nucleotide 1849 in exon 12 of the JAK2 gene, located on the chromosome 9p, leading to a substitution of valine to phenylalanine at amino acid position 617 in the JAK2 protein. The aim of this study was to assess the prevalence of JAK2 mutation in MPN patients.

Materials and Methods: In this experimental study we evaluated JAK2 mutation in 58 patients with MPNs by simple randomized sampling. The mutation was detected by ARMS-PCR in patients.

Results: The JAK2 V617F mutation was detected in 86.6% (26/30) of patients with polycythemia Vera, 46.6% (7/15) of patients with essential thrombocythemia and 61.5% (8/13) of patients with idiopathic myelofibrosis. Polycythemia Vera patients carrying the mutation displayed a higher levels of WBC (p=0.03) and 61.5% (16/26) of these patients were females. The differences in other groups were not significant. The mutation was confirmed by sequencing.

Conclusions: Our Results show similarity with other studies. Thus, ARMS-PCR can be applied as differential diagnosis test for detection of JAK2 mutation in suspected patients with MPNs.

Background and Aim: The JAK2 is an acquired mutation that is observed in majority of patients with classical Philadelphia-negative Myeloproliferative neoplasms that include polycythemia vera (PV), essential thrombocythemia (ET), primary myelofibrosis (PMF). This acquired mutation is characterized by a G to T transversion at nucleotide 1849 in exon 12 of the JAK2 gene, leading to a substitution of valine to phenylalanine at amino acid position 617(V617F) of the JAK2 protein, and result in constitutive JAK2 activation that promotes hypersensitivity to growth factors and cytokines.

Materials and Methods: In this study we evaluated RNA from 58 patients with MPNs and statistical analysis was done with mann whitney test. The mutation detected by AS-PCR. In addition, 3 samples were sequenced in Mille gen company.

Results: 46 patients:86.6%(26/30) of those with polycythemia vera, 53.3% (8/15) of those with essential thrombocythemia,61.5% (8/13) of those with idiopathic myelofibrosis polycythemia vera patient carrying the mutation displayed higher levels of WBC (p=0.03). on the other hand,16 out of 26 JAK2V617F positive patients were female there is a demonstrate correlation between the presence of a mutant allele and female gender. The difference in other groups were not significant.

Discussion and Conclusion: The JAK2V617F mutation has been detected in the vast majority of patients with polycythmia vera (65-95%) and in a lower frequency in patients with essential thrombocythemia (23-57%), idiopathic myelofibrosis (23-57%) and chronic myeloid leukemia 19% (3/16 CML Ph-). Detection of the mutation is helpful in differential diagnosis, prognosis, and prediction of therapeutic response.

Background and Aim: Acute leukemia is one of the main causes of cancer in the world. Now a days using natural materials as source of anticancer drugs is more recommended. HESA-A is a drug of herbal-marine origin (patented by Iranian researcher). HESA-A is composed of 50% inorganic substance، 45% organic substance (aminoenthraquinone) and 5% water. In this study effects of HESA-A، on NB4 cell line (Acute promyelocytic leukemia cells) was evaluated.

Materials and Methods: HESA-A was prepared in normal saline as a stock solution (80 mg/ml, PH=7.4), and then was sterilized. After culturing and proliferation of NB4 cell line, the cells were treated by doses of 1, 2, 4 and 8 mg/ml of HESA-A. Respectively after 72h, the percentage of viable and dead cells were counted by using Trypan blue staining in Neubanr hemocytometer. Then by MTTassay, the percentage of cell survival were determined by ELISA reader in 570nm. Finally the effects of HESA-A on apoptosis were evaluated by flocytometery.

Results: This invitro study shows that HESA-A has a cytotoxcic and antiprolifrative effects against NB4 cell line (Dose dependent).IC50 dose was 5mg/ml .HESA-A can result in apoptosis in 50% of the cells.

Conclusion: Although the mechanism of HESA-A cytotoxicity action is not known, yet this study shows that this drug may cause apoptosis of cells by dose dependent method.

Background and Aim: Chronic myelogenous leukemia is characterized by Philadelphia (Ph) chromosome, the presence of BCR-ABL fusion gene and constitutive activation of the ABL1 tyrosine kinase. Despite an excellent result of target therapy by imatinib, some patients develop resistance to imatinib. In this study Efficacy of HESA-A on proliferation and apoptosis of K562 cell line was assessed.

Materials and Methods: In this study doubling time of K562 cell line was calculated. The cells were affected by various concentrations of HESA-A(1,2,4 and 8 mg/ml respectively). Cytotoxicity and IC50 dose of HESA-A were detected by MTT and trypan blue exclusion assay. Apoptosis was assessed by flowcytometry after 48 h cell treatment in the presence of IC50 dose.

Results: Doubling time of K562 cells was 24 hours. HESA-A reduced the number of viable K562 cells in a concentration dependent manner.IC50 dose was 3.5 mg/ml. In flowcytometry analysis of apoptosis, 19.22% of the treated cells were located in the position of the necrotic cells.

Conclusion: The results of MTT and trypan blue exclusion assay suggest that HESA-A inhibits the growth of k562 cells in a concentration dependent manner and induces necrosis in K562 cells.

Background and Aim: Chronic myeloid leukemia (CML) is a myeloprolifrative neoplasm that is characterized by an expansion of myeloid, erythroid cells and platelets in peripheral blood and myeloid hyperplasia in bone marrow. Secreted frizzled-related protein family is a negative regulator of the Wnt signaling pathway that suppresses this signaling pathway in healthy individuals. Aberrant regulation of the Wnt signaling pathway is a prevalent theme in cancer biology, and methylation in promoter of SFRP family has been shown to cause uncontrolled cell proliferation in cancer. Chronic myeloid leukemia was the first malignancy in which the important role of Wnt signaling pathway has been described. 
In the present study, we examined the methylation status of SFRP1 and SFRP2 genes in patients with CML.
Materials and Methods: Blood samples were obtained from 25 healthy individuals and 33 patients whit chronic meyloied leukemia (23 male, 10 female) Then Isolated DNA was treated with sodium bisulfite and analyzed by methylation-specific polymerase chain reaction (MSP) with primers specific for methylated and unmethylated promoter sequences of the SFRP1 & -2 genes. We used Mann-Whitney u-tests to investigate the correlation between SFRP-1 and SFRP-2 genes hypermethylation and clinical parameters.
Results: In CML patient hypermethyleation frequency of SFRP-1 and SFRP-2 genes were 16.1℅ and 27.2% respectively. In control group SFRP-1 and SFRP-2 genes were unmethylated.
Conclusion: The present study showed that, methylation of SFRP genes also occurs in CML like other solid tumors. Therefore, the methylation of these genes may play a role in the initiation of malignant disease.