MiRNA -101 transfection and its effect on the cytotoxicity induction and expression of ubiquitin ligase HECTH9 in acute myeloid leukemia cells (AML)

Abstract

Introdution: In the present study, Lipofactamine 2000 was used as a cationic liposome for miR-101 transfection in order to investigate its cytotoxicity and its effect on the expression of ubiquitin ligase HECTH9 in acute myeloid leukemia cells (AML). Methods: MiR- 101 was transferred to KG-1 cells (myeloid cells) and HBMF-SPH (healthy bone marrow cells) using lipofectamine 2000 as a nano carrier. Then, using the MTT test, the 48-hour cell toxicity in both cell lines was evaluated. The effect of this miRNA on the expression of HECTH9 gene (ubiquitin Ligaase E3) was evaluated using qRT-PCR technique. Results: The findings of this study showed that Lipofactamine alone was not toxic to any of the cell lines, but lipofectamine-containing miR-101 (Lipo / miR-101) in KG-1 cells produced the highest toxicity compared to other treatments. The results of qRT-PCR test showed that Lipo / miR-101 treatment in KG-1 cells caused the highest expression in HECTH9 gene at the mRNA level.   Conclusion: Lipofactamine, as a cationic liposome, can effectively transfect miR-101 into the cells and can cause miR-101 to specifically display its antitumor effect by increasing the expression of HECTH9 and regulating pathways of mitochondrial apoptotic pathway. Therefore, miR-101 can be used as a potent tumor suppressor and an effective therapeutic agent for gene therapy in the patients with AML.

Keywords

• lipofectamine 2000
• miR-101
• HECTH9
• gene therapy
• AML