Targeting folate receptors (α1) to internalize the bleomycin loaded DNA-nanotubes into prostate cancer xenograft CWR22R cells

Baig, Mirza Muhammad Faran Ashraf and Lai, Wing-Fu and Akhtar, Muhammad Furqan and Saleem, Ammara and Mikrani, Reyaj and Farooq, Muhammad Asim and Ahmad, Saud Asif and Tahir, Anum and Naveed, Muhammad and Abbas, Muhammad and Ansari, Muhammad Tayyab (2020) Targeting folate receptors (α1) to internalize the bleomycin loaded DNA-nanotubes into prostate cancer xenograft CWR22R cells. Journal of Molecular Liquids.

Text (Research Article) 05 Targetting Folatereceptors Baig et al.pdf - Published Version Restricted to Registered users only Download (3MB) | Request a copy

Abstract

DNA-nanotechnology based on DNA scaffolding technique is an established approach to formulate water miscible
nano-structural frameworks. We have designed the hydrophilic DNA-nanotubes (D-NTs) as a (water soluble)
vehicle to encapsulate a water-insoluble DNA-intercalating anticancer drug. Bleomycin (BM) is a
model hydrophobic anticancer drug used in this study capable to bind with the D-NTs to formulate BM loaded
D-NTs. This BM@D-NTs system was evaluated for the improvement of the in vitro anti-cancer effects on the resistant prostate cancer xenograft CWR22R cells, over-expressed with the folate receptors/alpha (FRα). D-NTs were
functionalized with the FRα-targeting antibodies to interact with the FRα (receptors) highly expressed on the
resistant prostate cancer xenograftCWR22R cells. D-NTs not only increased the aqueous miscibility/dispersibility
of BM but also enhanced the therapeutic efficiency as a targeted (site-specific) drug delivery system. D-NTs synthesis was achieved by sticky ends cohesions of DNA triangular tiles. DNA triangles were self-assembled from a
freshly circularized short scaffold chain (84-NT) by annealing with the various staple strands. The polymerization of the triangular tiles gave rise to DNA-nanosheet lattices which underwent morphological transition guided by
the twists in the DNA duplexes. This DNA double helix curvature caused self-coiling of the DNA 2D nano-sheets to
condense into D-NTs morphology. Native-Page gel experiment showed decreased electrophoretic mobility down
the gel confirming the successful execution of the polymerized lattices via sticky ends cohesion of the DNA triangles.
The final morphology and self-coiling of 2DDNA nano-sheets were confirmed through atomic force microscopy
(AFM) showed the successful synthesis of D-NTs having diameter 3 to 5 μm and length 200 to 600 nm.
BM was loaded onto D-NTs via incubation of hydro-alcoholic solution of the BM with the aqueous solution of DNTs
followed by the evaporation of alcohol and intercalation of the BM onto D-NTs. The intercalation of BM onto
D-NTs was confirmed by UV-shift analysis. The targeted cytotoxicity of the BM@D-NTs for the CWR22R (resistant
prostate cancer xenograft) cells was confirmed through MTT assay and the flow cytometry compared to the
highly compatible empty D-NTs. Confocal microscopy revealed the time-dependent transfection of BM@D-NTs into CWR22R cells compared to the control cell line.

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