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Formulation of Stattic as STAT3 inhibitor in nanostructured lipid carriers (NLCs) enhances efficacy of doxorubicin in melanoma cancer cells

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Abstract

Nowadays, nanoparticle-based combination therapy has been emerging as huge innovation in cancer treatment. Here, we studied the effect of Stattic (STAT3 inhibitor) loaded in nanostructured lipid carriers (NLCs) on enhancing the efficacy, cytotoxicity, and induction of apoptosis of doxorubicin in B16F10 mouse melanoma cancer cell. The evaluation of Stattic-loaded NLCs has been done in terms of zeta potential, particle size, scanning electron microscope (SEM), and cellular uptake. MTT assay was applied to evaluate the cell proliferation. Apoptotic cell death and identification of early and late apoptosis were assessed by DAPI staining and Annexin V/PI staining, respectively. Real-time RT-PCR was applied to measure the effects of doxorubicin and/or Stattic on key apoptotic genes such as Bad, Survivin, HIF1, and STAT3. The Stattic formulated into NLCs shown mean particle size of 56 ± 7 nm which was confirmed by SEM. The IC50 values for Stattic and doxorubicin were 2.95 ± 0.52 μM and 1.21 ± 0.36 μM, respectively. Stattic-loaded NLCs diminished percent of cell proliferation from 68 ± 6.8 to 54 ± 3.7% (p < 0.05). Combinational treatment of the cells with Stattic-loaded nanoparticles and doxorubicin give rise to a significant increase in the percentage of apoptosis (p < 0.05). The study of gene expression profile has shown a remarkable decrease in anti-apoptotic gene, Survivin, along with smooth decline in HIF1 as angiogenesis intermediator and increase in Bad mRNA levels. Our results recommend that NLCs as novel technology have potent strategy to augment efficacy of current chemotherapeutic agent in melanoma cancer cells.

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Funding

The authors received support from the “Drug Applied Research Center, IR.TBZMED.REC. 1396.308.”

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Authors and Affiliations

  1. Drug Applied Research Center, and Department of Medical Biotechnology, Tabriz University of Medical Sciences, Tabriz, Iran

    Jamal Mohammadian, Shiva Mahmoudi, Fatemeh Salehnia, Nazila Fathi Maroufi & Nasser Samadi

  2. Department of Clinical Biochemistry, School of Medicine, Ardabil University of Medical Science, Ardabil, Iran

    Shiva Mahmoudi & Pirouz Pourmohammad

  3. Department of Genetic and Bioengineering, Faculty of Engineering and Architecture, Yeditepe University, 34755, Istanbul, Turkey

    Mohammad Pirouzpanah

  4. Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, Canada

    Mehdi Sabzichi

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  1. Jamal Mohammadian
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  2. Shiva Mahmoudi
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Contributions

J.M and SH.M had the main contribution in conception and design and are involved in the performance of experiments, data analysis, and interpretation. P.P manuscript writing and editing and revision; M.P and F.S are involved in the data analysis and manuscript writing; N.F are involved in some molecular experiments; N.S and M.S. are involved in the performance of all experiments. We stated that our team did not use a paper mill and performed all experiments in-house.

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Correspondence to Nasser Samadi or Mehdi Sabzichi.

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Mohammadian, J., Mahmoudi, S., Pourmohammad, P. et al. Formulation of Stattic as STAT3 inhibitor in nanostructured lipid carriers (NLCs) enhances efficacy of doxorubicin in melanoma cancer cells. Naunyn-Schmiedeberg's Arch Pharmacol 393, 2315–2323 (2020). https://doi.org/10.1007/s00210-020-01942-x

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