Volume 30, Issue 6 (September 2019)
Studies in Medical Sciences 2019, 30(6): 475-486 |
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Abedi Gaballu F, Mansoori B, Dehghan G. STUDY OF HMGA2 GENE INHIBITION WITH SPECIFIC SHRNA AND SIRNA AND INVESTIGATION OF CORRESPONDING EFFECTS ON DOWNSTREAM GENE EXPRESSION IN MDA-MB-231 CANCER CELLS: A BIOINFORMATIC AND EXPERIMENTAL STUDY. Studies in Medical Sciences 2019; 30 (6) :475-486
URL: http://umj.umsu.ac.ir/article-1-4753-en.html
URL: http://umj.umsu.ac.ir/article-1-4753-en.html
Professor, Department of Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran. , dehghan2001d@yahoo.com
Abstract: (3369 Views)
Background & Aims: The use of siRNA to silence gene expression is increasingly expanding today. The aim of this study is to bioinformatically and experimentally investigate the inhibition of the HMGA2 gene and its corresponding effects on downstream genes expression rate in MDA-MB-231 cancer cell treated by shRNA and siRNA specific to HMGA2.
Materials & Methods: To perform this bioinformatic and experiment study, first microarray data were collected from Gene Expression Omnibus (GEO) and then analyzed by Probabilistic neural networks (PNN) in MATLAB 2018a software as a bioinformatics tool. Next, the HMGA2 siRNA was designed and obtained. SiRNA transfection was carried out using lipofectamine as a carrier. The expression of HMGA2 gene, oncogene, and tumor suppressor genes were evaluated by real-time PCR.
Results: The bioinformatics result revealed that HMGA2 gene can nearly correlate with downstream genes (oncogene or tumor suppressor genes). Transfection of siRNA into MDA-MB231 cancer cells significantly (p< 0.05) inhibits HMGA2 gene expression in comparison with the control group. In addition, following HMGA2 gene suppuration, the oncogene (TERT) and tumor suppressor gene (DEDD) expressions were significantly (p< 0.05) reduced and increased, respectively.
Conclusion: The HMGA2 gene due to wide correlations with oncogenes and tumor suppressor genes is a reasonable option for targeting and silencing specific siRNA. The successful inhibition of HMGA2 gene expression and impressing of TERT and DEDD expressions after transferring specific siRNA finding is in accordance with bioinformatics results.
Materials & Methods: To perform this bioinformatic and experiment study, first microarray data were collected from Gene Expression Omnibus (GEO) and then analyzed by Probabilistic neural networks (PNN) in MATLAB 2018a software as a bioinformatics tool. Next, the HMGA2 siRNA was designed and obtained. SiRNA transfection was carried out using lipofectamine as a carrier. The expression of HMGA2 gene, oncogene, and tumor suppressor genes were evaluated by real-time PCR.
Results: The bioinformatics result revealed that HMGA2 gene can nearly correlate with downstream genes (oncogene or tumor suppressor genes). Transfection of siRNA into MDA-MB231 cancer cells significantly (p< 0.05) inhibits HMGA2 gene expression in comparison with the control group. In addition, following HMGA2 gene suppuration, the oncogene (TERT) and tumor suppressor gene (DEDD) expressions were significantly (p< 0.05) reduced and increased, respectively.
Conclusion: The HMGA2 gene due to wide correlations with oncogenes and tumor suppressor genes is a reasonable option for targeting and silencing specific siRNA. The successful inhibition of HMGA2 gene expression and impressing of TERT and DEDD expressions after transferring specific siRNA finding is in accordance with bioinformatics results.
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