Combination of Nanovectorized siRNA Directed against Survivin with Doxorubicin for Efficient Anti-Cancer Activity in HER2+ Breast Cancer Cells
According to Globocan 2020, breast cancer is considered one of the most common cancers affecting women and is one of the leading causes of death in over 100 countries. The available classical treatment options do not always give satisfactory outcomes, and some patients develop resistance to these tr...
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| Format: | Article |
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MDPI AG
2022-11-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/14/11/2537 |
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| author | Sahar Eljack Emilie Allard-Vannier Yoann Misericordia Katel Hervé-Aubert Nicolas Aubrey Igor Chourpa Areeg Faggad Stephanie David |
| author_facet | Sahar Eljack Emilie Allard-Vannier Yoann Misericordia Katel Hervé-Aubert Nicolas Aubrey Igor Chourpa Areeg Faggad Stephanie David |
| author_sort | Sahar Eljack |
| collection | DOAJ |
| description | According to Globocan 2020, breast cancer is considered one of the most common cancers affecting women and is one of the leading causes of death in over 100 countries. The available classical treatment options do not always give satisfactory outcomes, and some patients develop resistance to these treatments. This study aims to investigate the combination of nanovectorized siRNA directed against anti-apoptotic protein Survivin (siSurvivin) by targeted stealth magnetic siRNA nanovectors (TS-MSN), designed in our lab, with Doxorubicin (DOX), as an option for HER2+ breast cancer treatment. The hypothesis is that the pretreatment of the HER2+ breast cancer cell line SK-BR-3 with siSurvivin will induce apoptosis in the cancer cells and enhance the therapeutic efficacy of DOX, allowing a dose reduction of DOX and hence a reduction of potential side effects. TS-MSN are based on superparamagnetic iron oxide nanoparticles (SPIONs) covalently coupled with a fluorophore sulfocyanine-5 and polyethylene glycol 5000 (PEG<sub>5000</sub>) and functionalized with single-chain variable fragments (scFv) of an antibody targeting the HER2 membrane receptor. These covalently functionalized SPIONs are then complexed via electrostatic interactions with therapeutic siRNA and the cationic polymers, chitosan, and poly-L-arginine. TS-MSN<sub>siSurvivin</sub> had an average size of 144 ± 30 nm, a PDI of 0.3, and a slightly positive zeta potential value of 10.56 ± 05.70 mV. The agarose gel electrophoresis assay confirmed that the siRNA is well-complexed into TS-MSN without leakage, as no free siRNA was detected. Moreover, siRNA in TS-MSN was protected from RNAse A degradation for up to 6 h at 37 °C. Formulations of TS-MSN with siSurvivin demonstrated in vitro gene knockdown up to 89% in the HER2+ breast cancer cell line SK-BR-3. Furthermore, qRT-PCR confirmed a significant Survivin mRNA relative expression inhibition (about 50%) compared to control siRNA or untreated cells. A combination protocol was evaluated between TS-MSN and Doxorubicin (DOX) for the first time. Therefore, SK-BR-3 cells were pretreated with TS-MSN formulated with siSurvivin at 50 nM for 24 h alone, before a DOX treatment at a concentration of 0.5 µM (corresponding to the IC<sub>50</sub>) was added for 48 h. The MTT cytotoxicity tests, performed after 72 h of treatment, revealed that the combination had a significant synergistic cytotoxic effect on SK-BR-3 cells compared to monotherapies or untreated cells. We confirmed that pretreatment of cells with siSurvivin potentializes the cytotoxic effect of DOX as an alternative approach for treating HER2+ breast cancer. In conclusion, a combination of anti-Survivin siRNA and DOX would be a good alternative in HER2+ breast cancer therapy. |
| first_indexed | 2024-03-09T18:03:36Z |
| format | Article |
| id | doaj.art-0afbba0fe3664611aecaf245fbd86a57 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-09T18:03:36Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
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| series | Pharmaceutics |
| spelling | doaj.art-0afbba0fe3664611aecaf245fbd86a572023-11-24T09:37:26ZengMDPI AGPharmaceutics1999-49232022-11-011411253710.3390/pharmaceutics14112537Combination of Nanovectorized siRNA Directed against Survivin with Doxorubicin for Efficient Anti-Cancer Activity in HER2+ Breast Cancer CellsSahar Eljack0Emilie Allard-Vannier1Yoann Misericordia2Katel Hervé-Aubert3Nicolas Aubrey4Igor Chourpa5Areeg Faggad6Stephanie David7EA6295 Nanomédicaments et Nanosondes, Université de Tours, 37200 Tours, FranceEA6295 Nanomédicaments et Nanosondes, Université de Tours, 37200 Tours, FranceEA6295 Nanomédicaments et Nanosondes, Université de Tours, 37200 Tours, FranceEA6295 Nanomédicaments et Nanosondes, Université de Tours, 37200 Tours, FranceISP UMR 1282, INRAE, Equipe BioMAP, Université de Tours, 37200 Tours, FranceEA6295 Nanomédicaments et Nanosondes, Université de Tours, 37200 Tours, FranceDepartment of Molecular Biology, National Cancer Institute University of Gezira (NCI-UG), Wad Medani 21111, SudanEA6295 Nanomédicaments et Nanosondes, Université de Tours, 37200 Tours, FranceAccording to Globocan 2020, breast cancer is considered one of the most common cancers affecting women and is one of the leading causes of death in over 100 countries. The available classical treatment options do not always give satisfactory outcomes, and some patients develop resistance to these treatments. This study aims to investigate the combination of nanovectorized siRNA directed against anti-apoptotic protein Survivin (siSurvivin) by targeted stealth magnetic siRNA nanovectors (TS-MSN), designed in our lab, with Doxorubicin (DOX), as an option for HER2+ breast cancer treatment. The hypothesis is that the pretreatment of the HER2+ breast cancer cell line SK-BR-3 with siSurvivin will induce apoptosis in the cancer cells and enhance the therapeutic efficacy of DOX, allowing a dose reduction of DOX and hence a reduction of potential side effects. TS-MSN are based on superparamagnetic iron oxide nanoparticles (SPIONs) covalently coupled with a fluorophore sulfocyanine-5 and polyethylene glycol 5000 (PEG<sub>5000</sub>) and functionalized with single-chain variable fragments (scFv) of an antibody targeting the HER2 membrane receptor. These covalently functionalized SPIONs are then complexed via electrostatic interactions with therapeutic siRNA and the cationic polymers, chitosan, and poly-L-arginine. TS-MSN<sub>siSurvivin</sub> had an average size of 144 ± 30 nm, a PDI of 0.3, and a slightly positive zeta potential value of 10.56 ± 05.70 mV. The agarose gel electrophoresis assay confirmed that the siRNA is well-complexed into TS-MSN without leakage, as no free siRNA was detected. Moreover, siRNA in TS-MSN was protected from RNAse A degradation for up to 6 h at 37 °C. Formulations of TS-MSN with siSurvivin demonstrated in vitro gene knockdown up to 89% in the HER2+ breast cancer cell line SK-BR-3. Furthermore, qRT-PCR confirmed a significant Survivin mRNA relative expression inhibition (about 50%) compared to control siRNA or untreated cells. A combination protocol was evaluated between TS-MSN and Doxorubicin (DOX) for the first time. Therefore, SK-BR-3 cells were pretreated with TS-MSN formulated with siSurvivin at 50 nM for 24 h alone, before a DOX treatment at a concentration of 0.5 µM (corresponding to the IC<sub>50</sub>) was added for 48 h. The MTT cytotoxicity tests, performed after 72 h of treatment, revealed that the combination had a significant synergistic cytotoxic effect on SK-BR-3 cells compared to monotherapies or untreated cells. We confirmed that pretreatment of cells with siSurvivin potentializes the cytotoxic effect of DOX as an alternative approach for treating HER2+ breast cancer. In conclusion, a combination of anti-Survivin siRNA and DOX would be a good alternative in HER2+ breast cancer therapy.https://www.mdpi.com/1999-4923/14/11/2537siRNAtargeted nanovectorHER2+DoxorubicinSurvivin |
| spellingShingle | Sahar Eljack Emilie Allard-Vannier Yoann Misericordia Katel Hervé-Aubert Nicolas Aubrey Igor Chourpa Areeg Faggad Stephanie David Combination of Nanovectorized siRNA Directed against Survivin with Doxorubicin for Efficient Anti-Cancer Activity in HER2+ Breast Cancer Cells Pharmaceutics siRNA targeted nanovector HER2+ Doxorubicin Survivin |
| title | Combination of Nanovectorized siRNA Directed against Survivin with Doxorubicin for Efficient Anti-Cancer Activity in HER2+ Breast Cancer Cells |
| title_full | Combination of Nanovectorized siRNA Directed against Survivin with Doxorubicin for Efficient Anti-Cancer Activity in HER2+ Breast Cancer Cells |
| title_fullStr | Combination of Nanovectorized siRNA Directed against Survivin with Doxorubicin for Efficient Anti-Cancer Activity in HER2+ Breast Cancer Cells |
| title_full_unstemmed | Combination of Nanovectorized siRNA Directed against Survivin with Doxorubicin for Efficient Anti-Cancer Activity in HER2+ Breast Cancer Cells |
| title_short | Combination of Nanovectorized siRNA Directed against Survivin with Doxorubicin for Efficient Anti-Cancer Activity in HER2+ Breast Cancer Cells |
| title_sort | combination of nanovectorized sirna directed against survivin with doxorubicin for efficient anti cancer activity in her2 breast cancer cells |
| topic | siRNA targeted nanovector HER2+ Doxorubicin Survivin |
| url | https://www.mdpi.com/1999-4923/14/11/2537 |
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