Volume 26, Issue 1 (Apr 2018)
JSSU 2018, 26(1): 64-76 |
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Nikoonahad Lotfabadi N, Mohseni Kouchesfehani H, Sheikhha M H, Kalantar S M. MiRNA -101 transfection and its effect on the cytotoxicity induction and expression of ubiquitin ligase HECTH9 in acute myeloid leukemia cells (AML)
. JSSU 2018; 26 (1) :64-76
URL: http://jssu.ssu.ac.ir/article-1-4413-en.html
URL: http://jssu.ssu.ac.ir/article-1-4413-en.html
Narges Nikoonahad Lotfabadi 
, Homa Mohseni Kouchesfehani * , Mohammad Hasan Sheikhha , Seyed Mehdi Kalantar
, Homa Mohseni Kouchesfehani * , Mohammad Hasan Sheikhha , Seyed Mehdi Kalantar
Abstract: (6052 Views)
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.
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.
Type of Study: Original article |
Subject:
Genetics
Received: 2017/12/9 | Accepted: 2018/01/20 | Published: 2018/05/16
Received: 2017/12/9 | Accepted: 2018/01/20 | Published: 2018/05/16
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