6 research outputs found
Overexpression of MiR-506 in jurkat (Acute T cell leukemia) cell line
Background & Objective: Acute lymphoblastic leukemia (ALL) is a malignant disease that arises from various mutations in B or T-lymphoid progenitors. MicroRNAs (miRNAs) regulate gene expression by binding to the 3' untranslated region of proteincoding genes. Dysregulation of miRNA expression may result in the development of cancerous phenotypes. Therefore, for the first time in this field, the present study aims to investigate the effect of overexpression of miR-506 in Jurkat (acute T cell leukemia) cell line. Methods: In this study, Jurkat cell lines were cultured in RPMI-1640 medium. Next, miR-506 was transfected with concentrations of 50 and 100 nM with Lipofectamine 2000. The accuracy of the transfection was confirmed by the transfection of siRNA conjugated with FITC. 48 h after transfection, the cells were prepared for other tests (flow cytometry, MTT assay, and RNA extraction). The expression level of miR-506 in the cells was analyzed using the quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Finally, SPSS 21 software was used for the data analysis. Results: According to our results, the viability of cells in concentrations of 50 and 100 nM was significantly higher than the control group. By overexpression of miR-506, the expressions of pro-apoptotic genes (p53, p21) and anti-apoptotic gene B-cell lymphoma-2 (BCL-2) are decreased and increased, respectively. Conclusion: This study showed that miR-506 may function as an oncogenic miRNA in the T-ALL cell line. In conclusion, overexpression of miR-506 leads to an increase in viable cancer cells
Overexpression of microRNA-630 in Acute Leukemic T-cell line
Background: MicroRNAs (miRNAs) are noncoding RNAs that control the expression of their target mRNAs. It affects cancer cell proliferation and apoptosis as oncogenes or tumor suppressors. Dysregulation of miRNAs expression leads to the development of various cancers. Therefore, for the first time in this field, this study investigated the effect of overexpression of microRNA-630 on the Jurkat cells. Materials and methods:: In this experimental study, the Jurkat cells were divided into the four groups, i.e. non-transfected control group (A), scramble (B), transfected with 50 nM concentrations of miR-630 (C), and treated with 100 nM miR-630 (D). MiR-630 transfection was performed by lipofectamine 2000. Cancer cell growth in each group was analyzed with MTT assay. Flow cytometry investigated percent of viable, necrotic, and apoptotic cells. Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) measured the expression of P53, P21, and BCL2 genes. SPSS (version 21) especially Kruskal-Wallis and Mann-Whitney U tests were utilized for data analysis. Results: The results of MTT assay showed that the cell growth rates in C (118%) and D (136%) groups were significantly higher than that in the control group (P= 0.037 vs. 0.034). The percentage of early and late apoptosis in C (3.1% P=0.01, 4.2% P=0.02) and D (0.5% P=0.008, 0.4% P=0.006) groups were significantly lower than those in the control group. The expression of p53 and p21 in C (0.7 P=0.037, 0.62 P=0.034) and D (0.44 P=0.034, 0.53 P=0.038) Groups were significantly decreased compared with the control group. The expression of B-cell lymphoma-2 (Bcl2) in C (1.85) and D (3.26) groups were significantly increased compared with the control group (P= 0.037 vs. 0.024). Conclusion: Overexpression of miR-630 led induction of T-ALL cell growth and reduction of their apoptosis. These results emphasized that miR-630 contributed as an oncogenic microRNA in T-ALL cells