Preparation of 17-DMAG loaded β-cyclodextrin nanoparticles for targeting gene Hsp90 in breast cancer cells
Preparation of 17-DMAG loaded β-cyclodextrin nanoparticles for targeting gene Hsp90 in breast cancer cells
Hasan Mellatyar,1,*Sona Talaei,2
1. Department of Basic Sciences, Maragheh University of Medical Sciences, Maragheh, Iran 2. Department of Basic Sciences, Maragheh University of Medical Sciences, Maragheh, Iran
Introduction: Dysregulation of Hsp90 gene expression is found in breast cancer cells. Here we used β-cyclodextrin-17-dimethylaminoethylamino-17-demethoxy geldanamycin (17-DMAG) complexes and free 17-DMAG to inhibit the expression of the Hsp90 gene in the T47D breast cancer cells. The goal was to determine whether nanoencapsulation of 17-DMAG enhances the anti-cancer effects as compared to free 17-DMAG.
Methods: The double emulsion method was used to encapsulate 17-DMAG, and the properties of prepared complexes were characterized by H nuclear magnetic resonance (HNMR) and Fourier Transform Infrared (FTIR). Assessment of cytotoxicity of β-cyclodextrin nanoparticles loaded with 17-DMAG was done by the colorimetric cell viability (MTT) assay. After treatment of T47D cells with a determined amount of free 17-DMAG and 17-DMAG-loaded β-cyclodextrin, mRNA was extracted, and cDNA was synthesized. For assessing the expression of the Hsp90 gene, real-time PCR was performed.
Results: Taking into account 17-DMAG load, IC50 was meaningfully decreased in nanocapsulated 17-DMAG in comparison with free 17-DMAG. This finding was related to a decrease in Hsp90 gene expression.
Conclusion: 17-DMAG- β-cyclodextrin complexes are more effective than free 17-DMAG in down-regulating of Hsp90 expression, at the same time exert more great inhibitory effects. Therefore, β-cyclodextrin can be a superior carrier for this type of hydrophobic agent.
Keywords: Breast Cancer, Hsp90, 17-DMAG, β-cyclodextrin, Nanoparticle.