Detection of N-RAS gene mutations in codons 12,13 and 61 in patients with pediatric acute lymphoblastic leukemia

Background: Acute leukemia is the prevalent malignancy in pediatrics. One of the most important causing factors of acute lymphoblastic leukemia is mutations of proto oncogenes and their chang in to oncogenes. Activation of N-RAS proto-oncogene due to point mutations plays a major role in ALL malignancy. Since there was no report on the frequency of N-RAS gene mutation in Iranian pediatric ALL patients, therefore we decided to determine its frequency and compare the results with age, sex and type of ALL. Materials and Methods: In this study, 60 pediatric ALL patients from Tehran Mahak hospital were screened for the mutations of N-RAS gene at codons 12 ,13 1nd 61.DNA was extracted from peripheral blood samples before the start of chemotherapy. The above mentioned codons were amplified by PCR and analyzed by restriction endonuclease enzymes. Results: We could detect mutations in 7 cases of 60(11.7%) patients. Most of the mutations were detected in males with an age less than 5 years old. The frequency of mutations for codons 12, 13 and 61 were 8.3%,3.3% and 1.7% respectively. Most of the mutations (71.4%) were found in c-ALL subtype. Conclusion: We detected mutations in 11.7% of our ALL patients. In general , frequency of the mutations that we found was in agreement with the results of other studies. However , to do study with more patients and wider range of age using a combination of PCR-RFLP and direct gene sequencing is highly recommended

Investigating the Relationship Between MiR210 Upregulation and Hemoglobin Gamma Chain Expression

Background and Aim: Micro RNAs are a group of small non-coding RNAs which play an important role in multiple processes such as proliferation, differentiation, apoptosis, and cancer. Recent studies indicate that mir-210 is overexpressed into erythroid linage during the differentiation of hematopoietic precursor. The main goal of the present study is to investigate the influence of mir-210 on the pattern of expression in hemoglobin gamma chain. Materials and Methods: First, K562 cell line was cultured in RPMI1640 media. Then, pre-miR-210 was transferred into K562 cell line by lipofectamin. Finally, the alterations in the pattern of gamma chain expression were analyzed in days 7 and 14 by RT-PCR and real time PCR technique. Results: It was demonstrated that the overexpression of mir-210 in K562 cell line would lead to a 25-fold increase in the expression of gamma chain in comparison with the control group. Data analysis revealed that the change in the pattern of hemoglobin gamma chain expression was meaningful (p<0.002). Conclusion: Based on these data, overexpression of mir-210 can lead to a significant increase in the production of gamma chain. Therefore, more studies in the field may reveal the fact that an increase in mir-210 can be a suitable goal in the improvement of sickle cell anemia and β-thalassemia