Evaluation the Effect of Methotrexate on Expression Changes of LncRNA CPEB2 in Jurkat E6.1 cell line
Evaluation the Effect of Methotrexate on Expression Changes of LncRNA CPEB2 in Jurkat E6.1 cell line
Arezoo Hassani,1Mahnaz Eskandari,2Golnaz Asaadi Tehrani,3,*Sina Mirza Ahmadi,4
1. Msc of Molecular Genetic Department of Genetics, Zanjan Branch, Islamic Azad University, Zanjan, Iran. 2. Msc of Molecular Genetic Department of Genetics, Zanjan Branch, Islamic Azad University, Zanjan, Iran. 3. Assistant professor of Molecular Genetics, Department of Genetics, Zanjan Branch, Islamic Azad University, Zanjan, Iran. 4. Assistant professor of Molecular Genetics Department of Genetics, Zanjan Branch, Islamic Azad University, Zanjan, Iran
Introduction: In acute lymphoblastic leukemia (ALL), early lymphoid pioneer cells multiply and take the place of healthy hematopoietic cells in the bone marrow. A purine analog called methotrexate is used to treat autoimmune disorders and leukemia. It has both immunosuppressive and anticancer properties. By boosting adenosine release, activating adenosine receptor A2a, and preventing the conversion of BH2 to BH4 in leukemia, methotrexate suppresses the activation of nuclear factor B (NF-B). This study set out to investigate the effects of methotrexate on the expression of the LncRNA CPEB2 in the acute lymphoblastic leukemia Jurkat E6.1 cell line.
Methods: In this study, methotrexate was prepared at 1and 10µM doses. After cell passage, the Jurkat E6.1 (T-ALL) was purchased from Pasteur Institute of Iran at the passage I was then treated with methotrexate at 48h with indicated concentrations. Then RNA extraction and cDNA synthesis were done and the expression changes of CPEB2 and the
Housekeeping GAPDH gene were evaluated by Real-Time PCR. Finally, the results of Real-
Time PCR were analyzed by Rest 2002 Software.
Results: our findings discovered that after 48 hours of treatment with methotrexate at 1µM, the expression of LncRNA CPEB2 was considerably lower than in non-methotrexate samples and compared with GAPDH. Conforming to the results, it has been found that a concentration of 1µM at 48h was the optimal dose and time. The expressions of LncRNA CPEB2 at the doses of 1 and 10µM at 48h were 0.721 and 1.521 respectively (P<0.001).
Conclusion: It may be inferred from the findings of the investigation into how the expression of CPEB2 changed while it was being treated with methotrexate that the concentration of 1µM was successful in suppressing CPEB2 expression. The findings revealed that methotrexate, at a concentration of 1 µM, which is the drug's maximum potency, caused a further drop in CPEB2 expression. For 48 hours, methotrexate significantly slowed down the expression of the CPEB2 oncogene, and at some of the investigated concentrations, this effect was statistically significant. Decreased gene expression at the concentration of 1µM
demonstrated that the effect of the drug was dependent on concentration and with increasing dose of the drug; a decreasing gene expression was evident. The result suggests that methotrexate has a high potential for controlling and treating cancers.