Journal of

Background and Aim: Chronic Myelogenous Leukemia (CML) is a clonal hematopoietic stem cell disorder characterized by a translocation between chromosome 9 and 22 called Philadelphia Chromosome. Telomerase- essential enzyme that adds telomeric repeats into the telomeres- maintains integrity of  chromosomal ends. Most normal somatic cells do not express hTERT (catalytic subunit of telomerase) but most neoplasia and cancer cells express it. In this study we evaluated the hTERT expression in patients with CML at different phases of the disease.

Materials and Methods: In this cross sectional study, 73 samples of 45 patients with CML were studied. Twenty six of samples were taken from patients in chronic phase before therapy and 26 samples three month after therapy. Nine samples were taken in accelerated phase and 12 in blastic phase. hTERT expression was studied by RT-PCR and the results were analyzed using SPSS 15.

Results: Thirty three (73%) of patients were male and 12 (23%) were female. Patients were divided into three age groups 17-29, 30-40 and 41-75 years. Of 73 samples, 43 samples (58.9%) were positive for hTERT and 30 samples (41.1%) were negative for this gene. In chronic phase (before therapy) 69.2% of the samples were PCR positive, but after therapy only 38.5% of samples were PCR positive. In accelerated and blastic phases, 55.6% and 83.3% of samples were PCR positive respectively. The hTERT positivity was differently significant (p<0.05) among different phases of the disease.

Conclusion: Significant difference between hTERT expression in different phases of CML disease can be used as a useful molecular marker for fallowing up, prognosis and disease progression after treatment

Background & Aim : Chronic myeloid leukemia (CML) is a disorder of pluripotential hematopoietic stem cell that is as a myeloproliferative disease and occurs in about 15 percent of all leukemia. Two cell cycle regulatory proteins that function as tumor suppressor are P16INK4A and P14ARF. The origin of these two proteins is a human INK4A-ARF gene locus that located on chromosome 9p21. P16INK4A control retinoblastoma (Rb) and P14ARF control with p53 thought negative feedback. The purposes of this study, this was that whether these genes are preferable use as a factor in prognosis and progression of disease.

Materials and Methods: This research was a Cross sectional study.  The expression of p16INK4A and p14ARF mRNA in about 73 peripheral bloods (PB) Samples were collected from 45 CML patients at different phases of disease were assayed by reverse transcriptase polymerase chain reaction (RT-PCR). 26 samples were from patients at chronic phase before any treatment, 26 samples 3 month after treatment with imatinib, 9 samples in accelerated phase and 12 samples in Blastic phase.

Results. From 45 patients with CML, 33 patients (73%) were men and 12 patients (27%) were women. About 26 samples (35%) were p16INK4A positive and 55 samples (75%) were p14ARF RT-PCR positive. This expression of the two genes at different phases of disease were not statistically significant (p>0.05).

Conclusion: High percentage of the CML patients expressed P14ARF and P16INK4A genes. The expression of these gene at different phases of disease (diagnosis, accelerate, and Blastic phases) was not statistically significant even though, the expression of these genes was higher after the treatment.  The increased expression of these genes was probably because of the Imatinib treatment.

Background and Aim: Bone sialoprotein (BSP) is a specific marker of osteoblastic differentiation. In this research, the effect of Zoledronic Acid on BSP expression and methylation status during osteoblastic differentiation of mesenchymal stem cells (MSCs) was evaluated.

Materials and Methods: In this experimental study, MSCs were isolated from human bone marrow. For osteogenic differentiation, hMSCs were pulse treated with zoledronic acid, and were incubated in osteogenic differentiation medium for 3 weeks. The DNA and RNA were extracted after the first, second and third weeks of culture and also from undifferentiated MSCs. After Sodium bisulfate (SBS) treatment, gene specific methylation analysis for BSP was carried out using Methylation Specific PCR technique.

Results: BSP expression was observed in osteoblastic differentiated cells whereas it was not seen in MSCs. MSP showed that BSP was unmethylated during osteoblastic differentiation.

Conclusion: BSP was expressed from the first week of differentiation. This confirms that zoledronic acid accelerates osteoblastic differentiation. Unmethylation status of BSP indicates that zoledronic acid does not have any effect on BSP methylation status. Other genetic or epigenetic mechanisms may control BSP expression during osteoblastic differentiation induction by zoledronic acid.