Downregulation Of Skp2 Via Sirna Mediated Gene Knockdown And Its Effects On Foxo3 And C-Myc Expression In Aml T(8,21) Cells
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Date
2017-06
Authors
Purushodman, Kasturi
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Publisher
Universiti Sains Malaysia
Abstract
Acute Myeloid Leukemia (AML) is a hematopoitic stem cell disorder causes by inability of the cells to differentiate and proliferate in normal manner. S-phase kinase–associated protein 2 (SKP2) protein is highly responsible in recognizing and determining the degradation of target protein and is well known for its oncogene property as well as for its therapeutic target in various cancers. FOXO3a protein was identified to present in AML and acts as tumor suppressor gene. c-MYC plays a key role as proto-oncogene and is recognized to induce AML. Numerous experiments showed correlation between SKP2 and FOXO3a as well SKP2 and c-MYC. However, there is no information available yet on relationship between SKP2, FOXO3a and c-MYC gene in AML with t(8;21) translocation. In this study, the correlation between expression level of SKP2, FOXO3a and c-MYC gene were identified. Kasumi-1 cell line was used as a model of AML with t(8;21). SKP2 downregulation via siRNA mediated gene knockdown was carried out to identify its effect on FOXO3a and c-MYC gene at three different time points which were 24, 48 and 72 hours. siRNAs were introduced into Kasumi-1 cell line via electroporation and the expression level of each gene were identified using real-time PCR. The result showed a successful knockdown of SKP2 at all different time points with more than 50% (p<0.05). However there was no significance difference between both FOXO3a and c-MYC expression levels with or without SKP2 knockdown. Yet, the expression of FOXO3a gene was identified to increase slightly from 24 hours to 72 hours after SKP2 knockdown. In order to obtain significant value, prolonged gene knockdown is essential.
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Keywords
Acute Myeloid Leukemia , causes by inability of the cells