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| Author(s): |
Manuela Garcia Laveli da Silva
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Faculdade de Medicina (FM/SBD) |
| Defense date: | 2017-02-20 |
| Examining board members: |
Durvanei Augusto Maria;
Solange Castro Afeche;
Sandra Coccuzzo Sampaio Vessoni
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| Advisor: | Durvanei Augusto Maria |
| Abstract | |
Synthetic Phosphoethanolamine (Pho-s) is a molecule analogous to cell membrane phospholipids with antitumor, anti-inflammatory and antiproliferative effects. Our research group has developed several in vitro and in vivo studies with Pho-s showing satisfactory results regarding its antitumor efficacy. In this project we evaluated the in vitro antitumor effects of Pho-s and the liposomal formulation DODAC/Pho-s in human breast adenocarcinoma line (MCF7). The effects of the Pho-s cytotoxicity on the MCF7 tumor cell line were evaluated by determining the cell viability by the MTT colorimetric test and the concentration of IC50% were 37.2; 25.8; 1.8 mM in the 24, 48 and 72 hour periods, respectively. With DODAC treatment in MCF7 cells the IC50% was 0.3 mM and the treatment of DODAC/Pho-s at IC50% of 1.8 mM after 24 hours of treatment. The production of lipoperoxides radicals was evaluated by the formation of TBARS in MCF7 cells, treated for 24, 48 and 72 hours with Pho-s, showing no significant difference in lipoperoxidation values. Changes in the cell population distributions in the cell cycle phases evaluated by flow cytometry showed an increase of the fragmented DNA after 48 hours and stop at the G2/M phase after 24 hours. Alterations in the cellular arrangements were evaluated by means of the labeling with the fluorochromes acridine-orange and rhodamine 123 using laser confocal microscopy, in which the change in morphology, membrane fragmentation and loss of the projection of cytoplasmic prolongations were observed. Senescent cell induction was evaluated by enzymatic activity with the ?-galactosidase enzyme, showing a decrease in senescent cells at the highest concentrations of treatment with Pho-s and DODAC/Pho-s. Several control markers and cell cycle progression, stress, angiogenesis and hormone receptors and mitochondrial potential were evaluated by flow cytometry. The apoptotic activity of breast tumor cells was evaluated by Annexin V/PI, showing an increase mainly of late apoptosis and cell proliferation assays by the CFSE-DA with the Pho-s by flow cytometry, resulting in significant decrease of cell proliferation. Treatment of MCF 7 human breast adenocarcinoma cells with Pho-s has been shown to be a phospholipid-type compound with promising antitumor potential (AU) | |
| FAPESP's process: | 14/02344-1 - Evaluation of antitumoral liposomal formulation DODAC with synthetic compound phosphoethanolamine in human breast tumor cells |
| Grantee: | Manuela Garcia Laveli da Silva |
| Support Opportunities: | Scholarships in Brazil - Master |