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ARTICLE
Induction of Apoptosis and Autophagy Using Ectopic DSCR1 Expression in Breast Cancer Cells
Medical Biotechnology Division, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
* Corresponding Author: Seyed Ahmad Aleyasin. Email: .
Molecular & Cellular Biomechanics 2018, 15(4), 215-227. https://doi.org/10.32604/mcb.2018.01813
Abstract
Down syndrome critical region 1 gene (DSCR1) is an anti-angiogenesis gene that inhibits the growth of tumor cells. In this study, the role of autophagy and apoptosis in DSCR1-induced cytotoxicity were investigated in MDA-MB-468 breast cancer cells. Lentivirus vector harboring DSCR1 (LV-DSCR1+) was constructed in HEK 293 cells and the optimal dosage of lentivirus vector for infection was determined by the MTT assay. After infection of cells using LV-DSCR1+, acridine orange and ethidium bromide staining was performed to investigation of apoptosis and autophagy. Expression of DSCR1 and marker genes for angiogenesis (VEGF), apoptosis (Bax and Bcl2) and autophagy (LC3 and Beclin) were determined by Real time PCR. The cellular morphological changes related to apoptosis and autophagy was happened after 48 hours of viral infection. Fragmented bright orange nucleuses and vacuoles were observed due to the cell apoptosis and autophagy after acridine orange and ethidium bromide staining. Upregulation of Bax, Lc3, DSCR1 and Beclin1 and downregulation of Bcl2 and VEGF was detected due to treatment with LV-DSCR1+. These results demonstrated that LV-DSCR1+ can induce apoptosis and autophagy, therefore suggesting that it may serves as an efficient tool to breast cancer treatment.Keywords
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