Synergistic Effect of the Association of BR2 Peptide with 2-Aminoethyl Dihydrogen Phosphate in Triple-Negative Breast Cancer Cells Association of BR2 with 2ADP on TNBC
Main Article Content
Keywords
triple-negative breast cancer, peptide, monophosphoester, synergism
Abstract
Background: Breast cancer is one of the most common diseases among women worldwide. The triple negative subtype is the most aggressive, with low tumor-free survival and the worst clinical evolution, requiring the development of more effective and targeted therapies. The present study investigated the in vitro pharmacological effects of the association of BR2 peptide with 2-aminoethyl dihydrogen phosphate (2-AEH2P) on MDA-MB-231 and 4T1 triple-negative breast cancer cells.
Methods: The physical-chemical analysis of the peptide was performed using the Heliquest software, the cell viability was assessed using the MTT colorimeter method and the predictive pharmacological effect was evaluated using the Synergy Finder software.
Results: The results showed the BR2 tumor penetration peptide and the 2-AEH2P+BR2 association significantly increased cytotoxicity in the MDA MB-231 and 4T1 tumor lines, without compromising the viability of the normal fibroblastic cells. The results also showed that depending on the time and concentration, a synergistic effect was observed for the association with tumor cells, with a therapeutic window between 0.8 and 50µm for MDA-MB-231 tumor cells in 48h.
Conclusion: The results demonstrated in vivo antitumor and antiproliferative efficiency for MDA-MB-231 and 4T1 tumor cells with low toxicity for normal fibroblast cells, with MDA MB-231 cells being more sensitive to treatments.
References
1. Siegel RL, Miller KD, Fuchs HE, Jemal A. Cancer statistics, 2022. CA Cancer J Clin. 2022;72(1):7-33. doi:10.3322/caac.21708
2. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68(6):394-424. doi:10.3322/caac.21492
3. INCA. Estimativa 2020 : Incidência de Câncer No Brasil.; 2019.
4. Sørlie T, Tibshirani R, Parker J, et al. Repeated observation of breast tumor subtypes in independent gene expression data sets. PNAS. 2003;100(14):8418-8423. www.stanford.edubreastcancer
5. Carey LA, Perou CM, Livasy CA, et al. Race, Breast Cancer Subtypes, and Survival in the Carolina Breast Cancer Study. American Medical Association. 2006;295(21):2492-2502. https://jamanetwork.com/
6. Torres MDT, Pedron CN, Araújo I, Silva PI, Silva FD, Oliveira VX. Decoralin Analogs with Increased Resistance to Degradation and Lower Hemolytic Activity. ChemistrySelect. 2017;2(1):18-23. doi:10.1002/slct.201601590
7. Alberici L, Roth L, Sugahara KN, et al. De Novo design of a tumor-penetrating peptide. Cancer Res. 2013;73(2):804-812. doi:10.1158/0008-5472.CAN-12-1668
8. de Sousa Cabral LG, Hesse H, Freire KA, et al. The BR2 peptide associated with 2-aminoethyl dihydrogen phosphate is a formulation with antiproliferative potential for a triple-negative breast cancer model. Biomedicine & Pharmacotherapy. 2022;153:113398. doi:10.1016/j.biopha.2022.113398
9. Luna ACDL, Santos Filho JRDA, Hesse H, Neto SC, Chierice GO, Maria DA. Modulation of pro-apoptotic effects and mitochondrial potential on B16F10 cells by DODAC/PHO-S liposomes. BMC Res Notes. 2018;11(1). doi:10.1186/s13104-018-3170-7
10. Ferreira AK, Meneguelo R, Neto SC, Chierice GO, Maria DA. Synthetic Phosphoethanolamine Induces Apoptosis Through Caspase-3Pathway by Decreasing Expression of Bax/Bad Protein and Changes CellCycle in Melanoma. Journal of Cancer Science & Therapy . 2011;3(3):53-59. Accessed April 23, 2022. https://pesquisa.bvsalud.org/portal/resource/pt/biblio-1064212
11. Ferreira AK, Meneguelo R, Pereira A, Filho OMR, Chierice GO, Maria DA. Synthetic phosphoethanolamine induces cell cycle arrest and apoptosis in human breast cancer MCF-7 cells through the mitochondrial pathway. Biomedicine and Pharmacotherapy. 2013;67(6):481-487. doi:10.1016/j.biopha.2013.01.012
12. Manuela Garcia Laveli da Silva, Luciana Bastianelli Knop, Durvanei Augusto Maria. Meclizine Chloridrate and Methyl-β-Cyclodextrin Associated with Monophosphoester Synthetic Phosphoethanolamine Modulating Proliferative Potential in Triple-Negative Breast Cancer Cells. Journal of Pharmacy and Pharmacology. 2019;7(7). doi:10.17265/2328-2150/2019.07.006
13. Manuela Garcia Laveli da Silva, Laertty Garcia de Sousa Cabral, Monique Gonçalves Alves, et al. 2-aminoethyl Dihydrogen Phosphate as a Modulator of Proliferative and Apoptotic Effects in Breast Cancer Cell Lines. Journal of Pharmacy and Pharmacology. 2021;9(3). doi:10.17265/2328-2150/2021.03.001
14. Conceição TO, Cabral LGS, Laveli-Silva MG, et al. New potential antiproliferative monophosphoester 2-aminoethyl dihydrogen phosphate in K-562 and K-562 MDR+ leukemia cells. Biomedicine and Pharmacotherapy. 2021;142. doi:10.1016/j.biopha.2021.112054
15. Conceição T de O, Laveli-Silva MG, Maria DA. Phosphomonoester Phosphoethanolamine Induces Apoptosis in Human Chronic Myeloid Leukemia Cells. Journal of Pharmacy and Pharmacology. 2019;7(7). doi:10.17265/2328-2150/2019.07.009
16. Gautier R, Douguet D, Antonny B, Drin G. HELIQUEST: A web server to screen sequences with specific α-helical properties. Bioinformatics. 2008;24(18):2101-2102. doi:10.1093/bioinformatics/btn392
17. Kamaly N, Xiao Z, Valencia PM, Radovic-Moreno AF, Farokhzad OC. Targeted polymeric therapeutic nanoparticles: Design, development and clinical translation. Chem Soc Rev. 2012;41(7):2971-3010. doi:10.1039/c2cs15344k
18. Blanco E, Shen H, Ferrari M. Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat Biotechnol. 2015;33(9):941-951. doi:10.1038/nbt.3330
19. Zhang X, Lin C, Lu A, et al. Liposomes equipped with cell penetrating peptide BR2 enhances chemotherapeutic effects of cantharidin against hepatocellular carcinoma. Drug Deliv. 2017;24(1):986-998. doi:10.1080/10717544.2017.1340361
20. Lim KJ, Sung BH, Shin JR, et al. A Cancer Specific Cell-Penetrating Peptide, BR2, for the Efficient Delivery of an scFv into Cancer Cells. PLoS One. 2013;8(6). doi:10.1371/journal.pone.0066084
21. Marqus S, Pirogova E, Piva TJ. Evaluation of the use of therapeutic peptides for cancer treatment. J Biomed Sci. 2017;24(1). doi:10.1186/s12929-017-0328-x
22. Huang W, Seo J, Willingham SB, et al. Learning from host-defense peptides: Cationic, amphipathic peptoids with potent anticancer activity. PLoS One. 2014;9(2). doi:10.1371/journal.pone.0090397
23. Mulder KCL, Lima LA, Miranda VJ, Dias SC, Franco OL. Current scenario of peptide-based drugs: the key roles of cationic antitumor and antiviral peptides. Front Microbiol. 2013;4. doi:10.3389/fmicb.2013.00321
24. Ferreira AK. Alquil Fosfatado Sintético Precursor Dos Fosfolipídios de Membrana Celular Com Potencial Efeito Antitumoral e Apoptótico Em Modelos de Tumores Experimentais.; 2013.
25. Ferreira AK, Meneguelo R, Pereira A, Filho OMR, Chierice GO, Maria DA. Anticancer Effects of Synthetic Phosphoethanolamine onEhrlich Ascites Tumor: An Experimental Study. Anticancer Res. 2012;32:95-104.
26. Ferreira AK, Meneguelo R, Pereira A, Filho OMR, Chierice GO, Maria DA. Synthetic phosphoethanolamine induces cell cycle arrest and apoptosis in human breast cancer MCF-7 cells through the mitochondrial pathway. Biomedicine and Pharmacotherapy. 2013;67(6):481-487. doi:10.1016/j.biopha.2013.01.012
27. Ferreira AK;, Meneguelo R;, Neto SC;, Chierice GO;, Maria DA. Synthetic Phosphoethanolamine Induces Apoptosis Through Caspase-3Pathway by Decreasing Expression OfBax/Bad Protein and Changes CellCycle in Melanoma. Vol 3.; 2011. https://pesquisa.bvsalud.org/portal/resource/pt/biblio-1064212
28. Ferreira AK, Santana-Lemos BAA, Rego EM, Filho OMR, Chierice GO, Maria DA. Synthetic phosphoethanolamine has in vitro and in vivo anti-leukemia effects. Br J Cancer. 2013;109(11):2819-2828. doi:10.1038/bjc.2013.510
29. Jabir MS, Taha AA, Sahib UI, Taqi ZJ, Al-Shammari AM, Salman AS. Novel of nano delivery system for Linalool loaded on gold nanoparticles conjugated with CALNN peptide for application in drug uptake and induction of cell death on breast cancer cell line. Materials Science and Engineering C. 2019;94:949-964. doi:10.1016/j.msec.2018.10.014
30. Lee YW, Hwang YE, Lee JY, Sohn JH, Sung BH, Kim SC. VEGF siRNA delivery by a cancer-specific cell-penetrating peptide. J Microbiol Biotechnol. 2018;28(3):367-374. doi:10.4014/jmb.1711.11025
31. Hoskin DW, Ramamoorthy A. Studies on anticancer activities of antimicrobial peptides. Biochim Biophys Acta Biomembr. 2008;1778(2):357-375. doi:10.1016/j.bbamem.2007.11.008
32. Shai Y. Mechanism of the binding, insertion and destabilization of phospholipid bilayer membranes by K-helical antimicrobial and cell non-selective membrane-lytic peptides. Biochimica et Biophysica Acta . 1999;1462:55-70. www.elsevier.com/locate/bba
33. Zhao J, Huang Y, Liu D, Chen Y. Two Hits Are Better than One: Synergistic Anticancer Activity of α-Helical Peptides and Doxorubicin/Epirubicin. Vol 6.; 2014. www.impactjournals.com/oncotarget/
34. Ning Zhang, Jia-Ning Fu, Ting-Chao Chou. Synergistic combination of microtubule targeting anticancer fludelone with cytoprotective panaxytriol derived from panax ginseng against MX-1 cells in vitro: experimental design and data analysis using the combination index method. Am J Cancer Res. 2016;6(1):97-104. www.combosyn.com
35. Bijnsdorp I v., Giovannetti E, Peters GJ. Analysis of Drug Interactions. In: ; 2011:421-434. doi:10.1007/978-1-61779-080-5_34
36. Koskimaki JE, Lee E, Chen W, et al. Synergy between a collagen IV mimetic peptide and a somatotropin-domain derived peptide as angiogenesis and lymphangiogenesis inhibitors. Angiogenesis. 2013;16(1):159-170. doi:10.1007/s10456-012-9308-7
37. Hu C, Chen X, Huang Y, Chen Y. Synergistic effect of the pro-apoptosis peptide kla-TAT and the cationic anticancer peptide HPRP-A1. Apoptosis. 2018;23(2):132-142. doi:10.1007/s10495-018-1443-1.
38. Bolhassani A, Jafarzade BS, Mardani G. In vitro and in vivo delivery of therapeutic proteins using cell penetrating peptides. Peptides (NY). 2017;87:50-63. doi:10.1016/j.peptides.2016.11.011.
2. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68(6):394-424. doi:10.3322/caac.21492
3. INCA. Estimativa 2020 : Incidência de Câncer No Brasil.; 2019.
4. Sørlie T, Tibshirani R, Parker J, et al. Repeated observation of breast tumor subtypes in independent gene expression data sets. PNAS. 2003;100(14):8418-8423. www.stanford.edubreastcancer
5. Carey LA, Perou CM, Livasy CA, et al. Race, Breast Cancer Subtypes, and Survival in the Carolina Breast Cancer Study. American Medical Association. 2006;295(21):2492-2502. https://jamanetwork.com/
6. Torres MDT, Pedron CN, Araújo I, Silva PI, Silva FD, Oliveira VX. Decoralin Analogs with Increased Resistance to Degradation and Lower Hemolytic Activity. ChemistrySelect. 2017;2(1):18-23. doi:10.1002/slct.201601590
7. Alberici L, Roth L, Sugahara KN, et al. De Novo design of a tumor-penetrating peptide. Cancer Res. 2013;73(2):804-812. doi:10.1158/0008-5472.CAN-12-1668
8. de Sousa Cabral LG, Hesse H, Freire KA, et al. The BR2 peptide associated with 2-aminoethyl dihydrogen phosphate is a formulation with antiproliferative potential for a triple-negative breast cancer model. Biomedicine & Pharmacotherapy. 2022;153:113398. doi:10.1016/j.biopha.2022.113398
9. Luna ACDL, Santos Filho JRDA, Hesse H, Neto SC, Chierice GO, Maria DA. Modulation of pro-apoptotic effects and mitochondrial potential on B16F10 cells by DODAC/PHO-S liposomes. BMC Res Notes. 2018;11(1). doi:10.1186/s13104-018-3170-7
10. Ferreira AK, Meneguelo R, Neto SC, Chierice GO, Maria DA. Synthetic Phosphoethanolamine Induces Apoptosis Through Caspase-3Pathway by Decreasing Expression of Bax/Bad Protein and Changes CellCycle in Melanoma. Journal of Cancer Science & Therapy . 2011;3(3):53-59. Accessed April 23, 2022. https://pesquisa.bvsalud.org/portal/resource/pt/biblio-1064212
11. Ferreira AK, Meneguelo R, Pereira A, Filho OMR, Chierice GO, Maria DA. Synthetic phosphoethanolamine induces cell cycle arrest and apoptosis in human breast cancer MCF-7 cells through the mitochondrial pathway. Biomedicine and Pharmacotherapy. 2013;67(6):481-487. doi:10.1016/j.biopha.2013.01.012
12. Manuela Garcia Laveli da Silva, Luciana Bastianelli Knop, Durvanei Augusto Maria. Meclizine Chloridrate and Methyl-β-Cyclodextrin Associated with Monophosphoester Synthetic Phosphoethanolamine Modulating Proliferative Potential in Triple-Negative Breast Cancer Cells. Journal of Pharmacy and Pharmacology. 2019;7(7). doi:10.17265/2328-2150/2019.07.006
13. Manuela Garcia Laveli da Silva, Laertty Garcia de Sousa Cabral, Monique Gonçalves Alves, et al. 2-aminoethyl Dihydrogen Phosphate as a Modulator of Proliferative and Apoptotic Effects in Breast Cancer Cell Lines. Journal of Pharmacy and Pharmacology. 2021;9(3). doi:10.17265/2328-2150/2021.03.001
14. Conceição TO, Cabral LGS, Laveli-Silva MG, et al. New potential antiproliferative monophosphoester 2-aminoethyl dihydrogen phosphate in K-562 and K-562 MDR+ leukemia cells. Biomedicine and Pharmacotherapy. 2021;142. doi:10.1016/j.biopha.2021.112054
15. Conceição T de O, Laveli-Silva MG, Maria DA. Phosphomonoester Phosphoethanolamine Induces Apoptosis in Human Chronic Myeloid Leukemia Cells. Journal of Pharmacy and Pharmacology. 2019;7(7). doi:10.17265/2328-2150/2019.07.009
16. Gautier R, Douguet D, Antonny B, Drin G. HELIQUEST: A web server to screen sequences with specific α-helical properties. Bioinformatics. 2008;24(18):2101-2102. doi:10.1093/bioinformatics/btn392
17. Kamaly N, Xiao Z, Valencia PM, Radovic-Moreno AF, Farokhzad OC. Targeted polymeric therapeutic nanoparticles: Design, development and clinical translation. Chem Soc Rev. 2012;41(7):2971-3010. doi:10.1039/c2cs15344k
18. Blanco E, Shen H, Ferrari M. Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat Biotechnol. 2015;33(9):941-951. doi:10.1038/nbt.3330
19. Zhang X, Lin C, Lu A, et al. Liposomes equipped with cell penetrating peptide BR2 enhances chemotherapeutic effects of cantharidin against hepatocellular carcinoma. Drug Deliv. 2017;24(1):986-998. doi:10.1080/10717544.2017.1340361
20. Lim KJ, Sung BH, Shin JR, et al. A Cancer Specific Cell-Penetrating Peptide, BR2, for the Efficient Delivery of an scFv into Cancer Cells. PLoS One. 2013;8(6). doi:10.1371/journal.pone.0066084
21. Marqus S, Pirogova E, Piva TJ. Evaluation of the use of therapeutic peptides for cancer treatment. J Biomed Sci. 2017;24(1). doi:10.1186/s12929-017-0328-x
22. Huang W, Seo J, Willingham SB, et al. Learning from host-defense peptides: Cationic, amphipathic peptoids with potent anticancer activity. PLoS One. 2014;9(2). doi:10.1371/journal.pone.0090397
23. Mulder KCL, Lima LA, Miranda VJ, Dias SC, Franco OL. Current scenario of peptide-based drugs: the key roles of cationic antitumor and antiviral peptides. Front Microbiol. 2013;4. doi:10.3389/fmicb.2013.00321
24. Ferreira AK. Alquil Fosfatado Sintético Precursor Dos Fosfolipídios de Membrana Celular Com Potencial Efeito Antitumoral e Apoptótico Em Modelos de Tumores Experimentais.; 2013.
25. Ferreira AK, Meneguelo R, Pereira A, Filho OMR, Chierice GO, Maria DA. Anticancer Effects of Synthetic Phosphoethanolamine onEhrlich Ascites Tumor: An Experimental Study. Anticancer Res. 2012;32:95-104.
26. Ferreira AK, Meneguelo R, Pereira A, Filho OMR, Chierice GO, Maria DA. Synthetic phosphoethanolamine induces cell cycle arrest and apoptosis in human breast cancer MCF-7 cells through the mitochondrial pathway. Biomedicine and Pharmacotherapy. 2013;67(6):481-487. doi:10.1016/j.biopha.2013.01.012
27. Ferreira AK;, Meneguelo R;, Neto SC;, Chierice GO;, Maria DA. Synthetic Phosphoethanolamine Induces Apoptosis Through Caspase-3Pathway by Decreasing Expression OfBax/Bad Protein and Changes CellCycle in Melanoma. Vol 3.; 2011. https://pesquisa.bvsalud.org/portal/resource/pt/biblio-1064212
28. Ferreira AK, Santana-Lemos BAA, Rego EM, Filho OMR, Chierice GO, Maria DA. Synthetic phosphoethanolamine has in vitro and in vivo anti-leukemia effects. Br J Cancer. 2013;109(11):2819-2828. doi:10.1038/bjc.2013.510
29. Jabir MS, Taha AA, Sahib UI, Taqi ZJ, Al-Shammari AM, Salman AS. Novel of nano delivery system for Linalool loaded on gold nanoparticles conjugated with CALNN peptide for application in drug uptake and induction of cell death on breast cancer cell line. Materials Science and Engineering C. 2019;94:949-964. doi:10.1016/j.msec.2018.10.014
30. Lee YW, Hwang YE, Lee JY, Sohn JH, Sung BH, Kim SC. VEGF siRNA delivery by a cancer-specific cell-penetrating peptide. J Microbiol Biotechnol. 2018;28(3):367-374. doi:10.4014/jmb.1711.11025
31. Hoskin DW, Ramamoorthy A. Studies on anticancer activities of antimicrobial peptides. Biochim Biophys Acta Biomembr. 2008;1778(2):357-375. doi:10.1016/j.bbamem.2007.11.008
32. Shai Y. Mechanism of the binding, insertion and destabilization of phospholipid bilayer membranes by K-helical antimicrobial and cell non-selective membrane-lytic peptides. Biochimica et Biophysica Acta . 1999;1462:55-70. www.elsevier.com/locate/bba
33. Zhao J, Huang Y, Liu D, Chen Y. Two Hits Are Better than One: Synergistic Anticancer Activity of α-Helical Peptides and Doxorubicin/Epirubicin. Vol 6.; 2014. www.impactjournals.com/oncotarget/
34. Ning Zhang, Jia-Ning Fu, Ting-Chao Chou. Synergistic combination of microtubule targeting anticancer fludelone with cytoprotective panaxytriol derived from panax ginseng against MX-1 cells in vitro: experimental design and data analysis using the combination index method. Am J Cancer Res. 2016;6(1):97-104. www.combosyn.com
35. Bijnsdorp I v., Giovannetti E, Peters GJ. Analysis of Drug Interactions. In: ; 2011:421-434. doi:10.1007/978-1-61779-080-5_34
36. Koskimaki JE, Lee E, Chen W, et al. Synergy between a collagen IV mimetic peptide and a somatotropin-domain derived peptide as angiogenesis and lymphangiogenesis inhibitors. Angiogenesis. 2013;16(1):159-170. doi:10.1007/s10456-012-9308-7
37. Hu C, Chen X, Huang Y, Chen Y. Synergistic effect of the pro-apoptosis peptide kla-TAT and the cationic anticancer peptide HPRP-A1. Apoptosis. 2018;23(2):132-142. doi:10.1007/s10495-018-1443-1.
38. Bolhassani A, Jafarzade BS, Mardani G. In vitro and in vivo delivery of therapeutic proteins using cell penetrating peptides. Peptides (NY). 2017;87:50-63. doi:10.1016/j.peptides.2016.11.011.
Article Sidebar
Article Details
How to Cite
1.
Garcia de Sousa Cabral L, Silva de Oliveira C, Cabette Barbosa Alves R, Oliveira Jr. VX, Poyet J-L, Augusto Mari D. Synergistic Effect of the Association of BR2 Peptide with 2-Aminoethyl Dihydrogen Phosphate in Triple-Negative Breast Cancer Cells: Association of BR2 with 2ADP on TNBC. Arch Breast Cancer [Internet]. 2023 Mar. 23 [cited 2023 Jun. 5];10(2):148-5. Available from: https://www.archbreastcancer.com/index.php/abc/article/view/650
Section
Original Article
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Copyright©. This is an open-access article distributed under the terms of the Creative Commons Attribution-Non-Commercial 4.0 International License, which permits copy and redistribution of the material in any medium or format or adapt, remix, transform, and build upon the material for any purpose, except for commercial purposes.
Article Statistics :Views : 106 | Downloads : 33 : 2