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Zoledronic acid-loaded lipidic nanoparticles enhance apoptosis and attenuate invasiveness by inhibiting epithelial to mesenchymal transition (EMT) in HepG2 cancer cells

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Abstract

The aim of this study was to evaluate the potential of zoledronic acid (ZOL)-loaded lipidic nanoparticles (ZOL-NLCs) in enhancing the efficiency of paclitaxel (Pac) in the context of cytotoxicity, apoptosis, and invasiveness of HepG2 hepatocellular carcinoma cells. ZOL-NLCs were characterized in terms of zeta potential, particle size, and scanning electron microscope (SEM) as well as cell internalization. To measure the anti-proliferative effects of ZOL-NLCs, annexin-V/PI and MTT assays were employed. Real-time PCR and western blot analysis were performed to identify the molecular mechanisms underlying the apoptosis in response to the studied conditions. Furthermore, the transwell migration assay was applied to clarify the role of applied formulations on the invasiveness of HepG2 cells. Our results demonstrated that the optimized ZOL had an average particle size of 105 ± 6 nm with a nearly narrow size distribution. The IC50 values for ZOL and ZOL-NLCs were 90 ± 3.1 and 54.6 ± 2.4 µM, respectively. The population of apoptotic cells was increased from 17 ± 2% to 27 ± 4% (p < 0.05) in response to treatment with ZOL-NLCs. ZOL-loaded nanoparticles triggered the mRNA expression of Bax as pro-apoptotic marker and E-cadherin as epithelial one along with a decrease in mesenchymal marker, N-cadherin, and Bcl-xl as an anti-apoptotic marker in HepG2 cells. These outcomes were consistent with western blot analysis of protein expressions. Besides, ZOL-incorporated lipidic nanoparticles reduced the migration of HepG2 cells significantly. Our data suggest that the formulation of ZOL into lipidic nanoparticles can be considered a potential therapeutic approach that can enhance the efficacy of Pac chemotherapy.

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Data is available as a supplementary file.

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Funding

This project was accomplished by a grant from the Liver and Gastrointestinal Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

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Authors and Affiliations

  1. Liver and Gastrointestinal Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

    Mehdi Sabzichi, Jamal Mohammadian, Alireza Mardomi & Fatemeh Ramezani

  2. Department of Immunology, Medical School, Rafsanjan University of Medical Sciences, Rafsanjan, Iran

    Omolbanin Oladpour

  3. Department of Pharmacology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran

    Mohsen Rashidi

  4. Faculty of Veterinary Medicine, Tabriz Branch Islamic Azad University, Tabriz, Iran

    Mahla Hosseinzadeh

  5. Department of Chemistry, Tabriz Branch Islamic Azad University, Tabriz, Iran

    Bahman Ramezani

  6. Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

    Marjan Ghorbani

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  1. Mehdi Sabzichi
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Contributions

M.S. and O.O. involved in the procedures of experiments. J.M., B.R., and M.H. had contribution in manuscript writing. M.R., M.G., and A.M. involved in performance molecular experiments and data analysis and interpretation. F.R. had the main contribution in conception, design, and manuscript editing as well as financial support. All authors read and approved the manuscript, and all data were generated in-house and that no paper mill was used.

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Correspondence to Fatemeh Ramezani.

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Mehdi Sabzichi1 and Omolbanin Oladpour contributed equally to this work.

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Sabzichi, M., Oladpour, O., Mohammadian, J. et al. Zoledronic acid-loaded lipidic nanoparticles enhance apoptosis and attenuate invasiveness by inhibiting epithelial to mesenchymal transition (EMT) in HepG2 cancer cells. Naunyn-Schmiedeberg's Arch Pharmacol 394, 2429–2439 (2021). https://doi.org/10.1007/s00210-021-02164-5

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