1. Knockdown of antiapoptotic genes in breast cancer cells by siRNA loaded into hybrid nanoparticles
- Author
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Tânia Beatriz Creczynski-Pasa, Adny Henrique Silva, Evelyn Winter, Leônidas João Mello, Gabriela Regina Rosa Souza, and Frederico Pittella
- Subjects
0301 basic medicine ,Small interfering RNA ,Materials science ,Cell Survival ,Cell ,bcl-X Protein ,Bioengineering ,Antineoplastic Agents ,Apoptosis ,Breast Neoplasms ,medicine.disease_cause ,Polyethylene Glycols ,03 medical and health sciences ,Gene Knockdown Techniques ,medicine ,Gene silencing ,Humans ,General Materials Science ,Electrical and Electronic Engineering ,RNA, Small Interfering ,Gene knockdown ,Mechanical Engineering ,General Chemistry ,Molecular biology ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,medicine.anatomical_structure ,Proto-Oncogene Proteins c-bcl-2 ,Mechanics of Materials ,Doxorubicin ,Cancer cell ,Cancer research ,MCF-7 Cells ,Nanoparticles ,Female ,Carcinogenesis - Abstract
Tumorigenesis is related to an imbalance in controlling mechanisms of apoptosis. Expression of the genes BCL-2 and BCL-xL results in the promotion of cell survival by inhibiting apoptosis. Thus, a novel approach to suppress antiapoptotic genes is the use of small interfering RNA (siRNA) in cancer cells. However, there are some limitations for the application of siRNA such as the need for vectors to pass the cell membrane and deliver the nucleic acid. In this study CaP-siRNA-PEG-polyanion hybrid nanoparticles were developed to promote siRNA delivery to cultured human breast cancer cells (MCF-7) in order to evaluate whether the silencing of antiapoptotic genes BCL-2 and BCL-xL by siRNA would increase cancer cell death. After 48 h of incubation the expression of BCL-2 and BCL-xL genes decreased to 49% and 23%, respectively. The siRNA sequence used induced cancer cell death at a concentration of 200 nM siRNA after 72 h of incubation. As the targeted proteins are related to the resistance to chemotherapeutic drugs, the nanocarriers systems were also tested in the presence of doxorubicin (DOX). The results showed a significant reduction in the CC50 of the DOX, after silencing the antiapoptotic genes. In addition, an increase in apoptotic cell counts for both incubations conditions was observed as well. In conclusion, silencing antiapoptotic genes such as BCL-2 and BCL-xL through the use of siRNA carried by hybrid nanoparticles showed to be effective in vitro, and presents a promising strategy for pre-clinical analysis, especially when combined with DOX against breast cancer.
- Published
- 2017