• Evaluation of the Effect of the Cyclophosphamide on Expression Changes of LncRNA HOXA-AS2 in Acute lymphoblastic leukemia
  • Neda Maleki khas,1 Shahine Mirzaei,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: The most typical kind of teenager cancer is leukemia. Acute lymphocytic leukemia (ALL), which makes up around 78% of all teenager leukemia diagnoses in this demographic, is diagnosed about five times more commonly than acute myelogenous leukemia (AML). One of the most successful and often used antineoplastic medications is still cyclophosphamide. Additionally, it has strong immunosuppressive properties and is the most widely used medication in bone marrow transplants (BMT). It was initially synthesized to selectively target cancer cells, although the hypothesized mechanism of tumor specificity (activation by cancer cell phosphamidases) transpired to be irrelevant to its activity. Nevertheless, cyclophosphamide's unique metabolism and inactivation by aldehyde dehydrogenase is responsible for its distinct cytotoxic properties. Differential cellular expression of aldehyde dehydrogenase has an effect on the anticancer therapeutic index and immunosuppressive properties of cyclophosphamide. Long noncoding RNAs (lncRNAs) have important roles in diverse cellular processes and carcinogenesis. Homeobox (HOX)A cluster antisense RNA 2 (HOXA-AS2) is a 1048-bp lncRNA located between human HOXA3 and HOXA4 genes whose overactivation was previously found to promote the proliferation and invasion of solid tumors. However, its biological roles in leukemia remain unclear. The goal of this study was to see how Cyclophosphamide affected the expression of the HOXA-AS2 gene in a cell line that had been diagnosed with acute lymphoblastic leukemia.
  • Methods: Two Cyclophosphamide concentrations were created for the current study: 20 and 50 µM at 48 hours. After purchasing the Jurkat E6.1 cell line from the Pasteur Institute, it was given a prepared dose of cyclophosphamide 48 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: The results of our findings showed that the expression of HOXA-AS2 in comparison with the GAPDH housekeeping gene decreased after 48hours of cyclophosphamide treatment at both of the concentration drug. According to the findings, changes in HOXA-AS2 gene expression decreased after 48 hours at a concentration of 1µM and 10µM decrease were statistically significant These changes included 20µM (0/958) and 50µM (0/923) at 48 hours, respectively. (P <0.001).
  • Conclusion: According to the present study results, alternation in HOXA-AS2 expression after treatment with cyclophosphamide, at two concentration were effective in decrease of HOXA-AS2 expression. Evidence showed that the cyclophosphamide has positive potential and efficacy because the drug was effective in decreasing gene expression in two concentrations in 48 hours. Therefore, cyclophosphamide can be a useful drug in controlling the expression of genes involved in acute lymphoblastic leukemia.
  • Keywords: LncRNA HOXA-AS2, GAPDH, Acute lymphoblastic leukemia, Cyclophosphamide.