Original Article
Assessing the selective therapeutic efficacy of superparamagnetic erlotinib nanoparticles in lung cancer by using quantitative magnetic resonance imaging and a nuclear factor kappa-B reporter gene system

https://doi.org/10.1016/j.nano.2018.01.010Get rights and content

Highlights

  • Dual functional FeDCE NPs may serve both therapeutic effect and diagnostic utility in the same time in NSCLC.

  • The cellular and tumoral internalization of FeDC-E NPs and the uptaken iron concentration can be estimated by MR ΔR2* values.

  • Lung tumor growth and NF-κB activity were markedly inhibited by FeDC-E NPs via the suppression of antiapoptotic effect.

  • On the contrary, FeDC-E NPs also effectively enhanced extrinsic and intrinsic apoptotic effects in lung cancer in vitro and in vivo.

Abstract

Non-small-cell lung cancer (NSCLC) is the most common type of lung cancer. Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors are commonly used as the first-line treatment for advanced NSCLC; however, the efficacy of drug delivery remains unknown. Hence, we successfully developed erlotinib-conjugated iron oxide nanoparticles (FeDC-E NPs) as theranostic probe that can potentially provide a new avenue for monitoring drug delivering through noninvasive magnetic resonance imaging. MRI ΔR2* relaxivity measurements offer an opportunity to quantitatively evaluate the uptake of FeDC-E NPs at cellular and tumoral levels. Additionally, NF-κB reporter gene system provides NF-κB activation status monitoring to validate the therapeutic efficiency of FeDC-E NPs. FeDC-E NPs not only inhibit the tumor growth and NF-κB-modulated antiapoptotic mechanism but also trigger extrinsic and intrinsic apoptotic pathways. Taken together, dual functional FeDC-E NPs offer diagnostic and therapeutic benefits against lung cancers, indicating that our presented probe could be applied in clinical.

Graphical Abstract

In this study, FeDCE NPs can be used as a probe to estimate the delivering and internalization in NSCLC by mMRI. Our FeDCE NPs potentially open a new avenue to identify the delivery efficacy and estimate concentration of drug by using mMRI (as shown in manuscript Figure 3, E). Moreover, FeDCE NPs induced apoptosis mechanism via inhibition of NF-κB mediated signaling pathway can be further validated by reporter gene system (as shown in manuscript Figure 6, B). In summary, this dual functional FeDCE NPs may serve both therapeutic effect and diagnostic utility in the same time in NSCLC.

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Section snippets

Cell line and culture

An EGFR-overexpressing human lung adenocarcinoma cell line, CL1-5-F4, was used in this study.20 Cells were kindly provided by Dr. Chia-Lin Hsieh (Taipei Medical University, Taiwan). CL1-5-F4 and CL1-5-F4/NF-κB-luc2 cells were cultured in Dulbecco’s modified Eagle’s medium and Ham’s F-12 nutrient medium supplemented with 10% fetal bovine serum (GE Healthcare Life Sciences HyClone Laboratories, UT, USA) and 1% penicillin–streptomycin (Invitrogen, Carlsbad, CA, USA). The cells were maintained in a

Treatment effects of FeDC-E NPs on CL1-5-F4/NF-κB-luc2 lung cancer cells

To evaluate the cytotoxicity of erlotinib within FeDC-E NPs, we performed the MTT assay. FeDC-E NPs were found to exhibit dose-dependent cytotoxicity against CL1-5-F4/NF-κB-luc2 cells, with an erlotinib IC50 value of approximately 12.8 μM (Figure 1, A). Different concentrations of free erlotinib toxicity were also validated using the MTT assay (Figure 1, A). In addition, no cytotoxicity was observed in the group that received FeDC NPs alone (Figure 1, B). Cytotoxicity was only found in the

Discussion

In this study, we evaluated the uptake of FeDC-E NPs and their effect on the NF-κB-modulated antiapoptotic mechanism through iron-sensitive T2* MRI, NF-κB molecular imaging, and other molecular techniques (flow cytometry, Western blotting, and IHC) in NSCLC in vitro and in vivo. Our previous study suggested that FeDC-E NPs inhibited the migration and invasion of tumor cells through ERK-NF-κB signaling; however, the effect of FeDC-E NPs on the apoptotic mechanisms remains unclear.19 Nelson et al

Author contribution

All the authors participated in the design, interpretation of the data, drafting and review of the manuscript.

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    Source of funding: We thank the Ministry of Science and Technology and Ministry of Health and Welfare of the Republic of China for their support (MOST 104-2314-B-038-051 MY3). The study was supported by Taipei Medical University and Taipei Medical University Hospital (grant nos. 104TMU-TMUH-23, TMU105-AE1-B49, 104TMUH-SP-02, 105TMUH-SP-01, and 105TMU-TMUH-23) and the Health and Welfare Surcharge on Tobacco Products (grant no. MOHW105-TDU-B-212-134001). This project was also supported by the National Health Research Institutes (MG-105-SP-07, MG-106-SP-07).

    Conflict of interest: All authors have no conflicts of interest to declare.

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