Evaluation the Effect of Methotrexate on Expression Changes of TUG1 in Acute Lymphoblastic Leukemia
Evaluation the Effect of Methotrexate on Expression Changes of TUG1 in Acute Lymphoblastic Leukemia
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. 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.