• Evaluation the Effect of Methotrexate on Expression Changes of TUG1 in Acute Lymphoblastic Leukemia
  • Arezoo Hassani,1 Mahnaz Eskandari,2 Golnaz 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. Acute lymphoblastic leukemia (ALL) is mainly a disease of childhood that arises from recurrent genetic insults that block precursor B and T cell differentiation and drive aberrant cell proliferation and survival. Recurrent defects including chromosomal translocations, aneuploidies, and gene-specific alterations generate molecular subgroups of B- and T-ALL with differing clinical courses and distinct responses to therapy. Recent discoveries arising from genome-wide surveys and adoptive transfer of leukemia-initiating cells have uncovered multiple gene copy number aberrations and have yielded new insight into at least one type of ALL-originating cell. Methotrexate, formerly known as amethopterin, is a chemotherapy agent and immune-system suppressant. It is used to treat cancer, autoimmune diseases, and ectopic pregnancies. Types of cancers it is used for include breast cancer, leukemia, lung cancer, lymphoma, gestational trophoblastic disease, and osteosarcoma A purine analog called methotrexate is used to treat autoimmune disorders and leukemia. It has both immunosuppressive and anticancer properties. in leukemia, methotrexate suppresses the activation of nuclear factor. This study set out to investigate the effects of methotrexate on the expression of the LncRNA TUG1 in acute lymphoblastic leukemia.
  • Methods: In this study, methotrexate was prepared at 1 and 10µM doses. After cell passage, the Jurkat E6.1 was prepared from Pasteur Institute of Iran at the passage I was then treated with methotrexate at 48 h with indicated concentrations. Then RNA extraction and cDNA synthesis were done and the expression changes of TUG1 and the Housekeeping GAPDH 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 48h of treatment with methotrexate at 1µM, the expression of LncRNA TUG1 was considerably lower than in non-methotrexate samples and compared with the reference gene. Conforming to the results, it has been shown of TUG1 at the concentration of 1 and 10µM at 48h were 0.523and 0.712 respectively (P <0.001).
  • Conclusion: as a conclusion investigation into how the expression of TUG1 changed while it was being treated with methotrexate the doses of 1 and 10µM were unsuccessful in suppressing TUG1 expression. The related results of methotrexate, at in 1 and 10µM, on TUG1 expression for 48h, showed down the expression of the TUG1 tumor suppressor. The result suggests that methotrexate doesn't have enough potential for controlling and treating cancers.
  • Keywords: Acute Lymphoblastic Leukemia, LncRNA, TUG , Methotrexate