Evaluation of the effect of the methotrexate on expression changes of LncRNA HOXA-AS2 in acute lymphoblastic leukemia
Evaluation of the effect of the methotrexate on expression changes of LncRNA HOXA-AS2 in acute lymphoblastic leukemia
Neda Maleki khas,1Shahine Mirzaei,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: One of the characteristics of acute lymphoblastic leukemia (ALL), a blood cancer of the lymphoid line of blood cells, is the development of a sizable number of immature lymphocytes. Pale skin, fever, easy bruising or bleeding, swollen lymph nodes, and bone soreness may all be symptoms in addition to weariness. ALL develops quickly as acute leukemia and, if ignored, usually results in death within a few weeks or months. Only 10% of adult ALL patients and 30% of pediatric ALL patients survive, making ALL a disease with a terrible prognosis. In cancer tissues, HOXA-AS2 is elevated, which promotes this type of cancer cell migration and proliferation. the finding also suggested that HOXA-AS2 levels may be significantly up-regulated in the bone marrow tissues of leukemia patients compared with healthy subjects. Furthermore, results showed that HOXA-AS2 upregulated HOXA3, thereby activating EGFR/Ras/Raf/MEK/ERK signaling pathway in leukemia. Methotrexate is used to treat certain types of cancer (such as acute lymphoblastic leukemia, non-Hodgkin's lymphoma) and etc. Another folate-dependent mechanism that is impacted by methotrexate is the methylation of biomolecules. Furthermore, independently of the metabolism of folate, methotrexate has the ability to alter metabolic pathways and cellular functions. The aim of this work was to investigate the effects of methotrexate on the expression of the HOXA-AS2 gene in an acute lymphoblastic leukemia cell line.
Methods: Methotrexate was produced in two concentrations for the current study: 1µM and 10µM at 72 hours. A prepared dose of methotrexate was administered to the Jurkat E6.1 cell line, which was bought from the Pasteur Institute, 72 hours after cell passage. Following RNA extraction and cDNA synthesis, Real-Time PCR was used to examine the changes in the expression of HOXA-AS2 and GAPDH. Finally, Excel was used to create the diagrams, and Rest 2002 Software to analyze the data.
Results: Our research found that after 72 hours of methotrexate treatment at both concentrations, the expression of HOXA-AS2 reduced in contrast to the GAPDH housekeeping gene. According to the findings, changes in HOXA-AS2 gene expression reduces after 72 hours at a concentration of 1µM and 10µM decrease were statistically significant These changes included 1µM (0/998) and 10µM (0/851) at 72 hours, respectively. (P <0.001).
Conclusion: According to the present study results, alternation in HOXA-AS2 expression after treatment with Methotrexate, at two concentration were effective in the decrease of HOXA-AS2 expression. Evidence showed that Methotrexate has positive potential and efficacy because the drug was effective in decreasing gene expression in two concentrations in 72 hours.