4aa44e5324c69ef3044d18a7eb2c09f8 pmc_33255217.pdf 79238bfcfdf819a1da9e1cb3f686bd6ac083d2fc pmc_33255217.pdf eeae149110a4a463e593681cec9fe112b482b48428b58711b38ff3c02f0bee8b pmc_33255217.pdf Title: Study and Preparation of Multifunctional Poly(L-Lysine)@Hyaluronic Acid Nanopolyplexes for the Effective Delivery of Tumor Suppressive MiR-34a into Triple-Negative Breast Cancer Cells Subject: Non-viral gene delivery using exogenous microRNAs is a potential strategy for fighting cancers with poor prognosis and which lack specific therapies, such as triple-negative breast cancer (TNBC). Herein we report the synthesis of six nontoxic electrostatic polymeric nanocapsules (P1 to P6) for microRNA delivery in TNBC cells. 1H Nuclear Magnetic Resonance (NMR) spectroscopy and Scanning Electron Microscopy (SEM) were used to characterize the nanopolyplexes, synthesized with Poly(L-Lysine) and hyaluronic acid (Ha). Studies on the activity of the ternary HA/PLI/miRNA-34 nanopolyplexes towards TNBC cell line MDA-MB-231 were conducted. The nanopolyplexes mediated intracellular restoration of tumor suppressor miR34a was evaluated by using Western blotting to quantify the expression level of the Bcl-2 protein. The results suggest that the P5, with a ratio PLI/Ha of 0.05, was the most promising for the delivery of miR-34a into TNBC cells; the P5 nanocapsules were able to reduce Bcl-2 expression at a protein level, and had an effect in the overall cell viability after 24 h treatment. Keywords: polyplexes RNA; Poly-Lysine; triple negative breast cancer Author: Jamila Djafari, Javier Fernández-Lodeiro, Hugo M. Santos, Julia Lorenzo, Sergi Rodriguez-Calado, Emilia Bértolo, José Luis Capelo-Martínez and Carlos Lodeiro Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.18 CreationDate: Thu Dec 3 08:32:07 2020 CET ModDate: Thu Dec 3 09:15:40 2020 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 9 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3045876 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- IOJCNK+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 YZONLH+VnURWPalladioL Type 1 Custom yes yes no 15 0 DXYPJC+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 EZRAFH+URWPalladioL-Bold Type 1 Custom yes yes no 24 0 WOZBHT+Rpxb Type 1 Builtin yes yes no 28 0 BYOEKM+URWPalladioL-Roma Type 1 Custom yes yes no 32 0 GJQQCT+Rpxr Type 1 Builtin yes yes no 36 0 YOTXCC+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 45 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 67 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 71 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 75 0 TGMEOR+Rpcxr Type 1 Builtin yes yes no 82 0 DFIRNB+Pxsy Type 1 Builtin yes yes no 86 0 SRTLFC+EURM10 Type 1 Builtin yes yes no 90 0 YWGLGW+Rpxi Type 1 Builtin yes yes no 97 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 109 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 113 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 117 0 Times New Roman TrueType WinAnsi no no no 121 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 124 0 ABCDEE+Calibri TrueType WinAnsi yes yes no 130 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 144 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 148 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 152 0 Times New Roman TrueType WinAnsi no no no 156 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 159 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 173 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 177 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 181 0 Times New Roman TrueType WinAnsi no no no 185 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 188 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 201 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 205 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 209 0 Times New Roman TrueType WinAnsi no no no 213 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 216 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 233 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 237 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 241 0 Times New Roman TrueType WinAnsi no no no 245 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 248 0 Times New Roman TrueType WinAnsi no no no 256 0 ABCDEE+Arial Bold,Bold TrueType WinAnsi yes yes no 258 0 Arial TrueType WinAnsi no no no 262 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 272 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 276 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 280 0 Times New Roman TrueType WinAnsi no no no 284 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 287 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-10-02 04:17:32 CEST RepresentationInformation: pmc_33255217.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-03-31 11:43:35 CEST Size: 3045876 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 400 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Synthesis of Imidazoled Poly-L-Lysine (PLI) Destination: subsection.2.1 Item: Title: Preparation and Characterization of Ternary Polyplexes HA/PLI/MiRNA-34 Destination: subsection.2.2 Item: Title: Scanning Electron Microscopy (SEM, Quanta650) Destination: subsection.2.3 Item: Title: Cell Cytotoxicity Assay Destination: subsection.2.4 Item: Title: Evaluation of the In Vitro Bcl-2 Suppression by MiR-34a Delivery Destination: subsection.2.5 Item: Title: Results and Discussion Destination: section.3 Item: Title: Conclusions Destination: section.4 Item: Title: References Destination: section.4 Info: Title: Study and Preparation of Multifunctional Poly(L-Lysine)@Hyaluronic Acid Nanopolyplexes for the Effective Delivery of Tumor Suppressive MiR-34a into Triple-Negative Breast Cancer Cells Author: Jamila Djafari, Javier Fernández-Lodeiro, Hugo M. Santos, Julia Lorenzo, Sergi Rodriguez-Calado, Emilia Bértolo, José Luis Capelo-Martínez and Carlos Lodeiro Subject: Non-viral gene delivery using exogenous microRNAs is a potential strategy for fighting cancers with poor prognosis and which lack specific therapies, such as triple-negative breast cancer (TNBC). Herein we report the synthesis of six nontoxic electrostatic polymeric nanocapsules (P1 to P6) for microRNA delivery in TNBC cells. 1H Nuclear Magnetic Resonance (NMR) spectroscopy and Scanning Electron Microscopy (SEM) were used to characterize the nanopolyplexes, synthesized with Poly(L-Lysine) and hyaluronic acid (Ha). Studies on the activity of the ternary HA/PLI/miRNA-34 nanopolyplexes towards TNBC cell line MDA-MB-231 were conducted. The nanopolyplexes mediated intracellular restoration of tumor suppressor miR34a was evaluated by using Western blotting to quantify the expression level of the Bcl-2 protein. The results suggest that the P5, with a ratio PLI/Ha of 0.05, was the most promising for the delivery of miR-34a into TNBC cells; the P5 nanocapsules were able to reduce Bcl-2 expression at a protein level, and had an effect in the overall cell viability after 24 h treatment. 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