Original Article
Sequential release of epigallocatechin gallate and paclitaxel from PLGA-casein core/shell nanoparticles sensitizes drug-resistant breast cancer cells

https://doi.org/10.1016/j.nano.2015.03.015Get rights and content

Abstract

Nanomedicines consisting of combinations of cytotoxic drugs and molecular targeted therapeutics which inhibit specific downstream signals are evolving as a novel paradigm for breast cancer therapy. This research addresses one such combination of Paclitaxel (Ptx), having several adversities related to the activation of NF-κB pathway, with Epigallocatechin gallate (EGCG), a multiple signaling inhibitor, encapsulated within a targeted core/shell PLGA-Casein nanoparticle. The sequential release of EGCG followed by Ptx from this core/shell nanocarrier sensitized Ptx resistant MDA-MB-231 cells to Ptx, induced their apoptosis, inhibited NF-κB activation and downregulated the key genes associated with angiogenesis, tumor metastasis and survival. More importantly, Ptx-induced expression of P-glycoprotein was repressed by the nanocombination both at the protein and gene levels. This combination also offered significant cytotoxic response on breast cancer primary cells, indicating its translational value.

From the Clinical Editor

Breast cancer is the most common cancer in women worldwide. As well as surgery, chemotherapy plays a major role in the treatment of breast cancer. The authors investigated in this article the combination use of a chemotherapeutic agent, Paclitaxel (Ptx), and an inhibitor of NF-?B pathway, packaged in a targeted nano-based delivery platform. The positive results provided a new pathway for future clinical use of combination chemotherapy in breast cancer.

Graphical Abstract

Combination nanomedicine of EGCG and Paclitaxel in a core-shell nanocarrier sensitizes drug-resistant breast cancer cells to low doses of Paclitaxel.

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Section snippets

Background

Breast cancer is the most common cancer that afflicts women worldwide and is also a major cause of cancer deaths among them.1 Chemotherapy is a widely accepted option for breast cancer. Of the various cytotoxic drugs administered, the taxane Paclitaxel (Ptx), is an extensively used antineoplastic drug exhibiting a single-agent overall response rate of 44-62%.2 However, several lines of evidence indicate that Ptx treatment can antagonize the therapy by inducing reactive oxygen species (ROS)

Materials and Methods

PLGA-Casein core/shell nanoparticles of 190 ± 12 nm particle size and surface charge − 41 ± 3.4 mV entrapping a combination of Ptx and EGCG at different doses in the core and shell respectively were prepared by an emulsion-precipitation route, as reported earlier.19 Targeted nanoparticles were developed using anti-EGFR and anti-HER2 antibodies for in vitro evaluation on MDA-MB-231 cells (anti-EGFR) and patient-derived tumor cells (anti-EGFR, anti-HER2). Refer to the supporting information for details.

Preparation and characterization of targeted dual drug-loaded core/shell nanoparticles

To prepare antibody-targeted drug-loaded nanoparticles, EDC-NHS coupling chemistry was employed, wherein the carboxyl groups on the surface of Casein shell were activated using EDC followed by stabilization with NHS. This yielded reactive –COO which interacts with the amine groups in the antibody to form an amide linkage (Figure S1 Supplementary Information). EGFR antibody conjugation efficiency was measured to be ~ 80% by spectrofluorimetry for an optimal antibody concentration of 200 ng/mL. It

Discussion

It is crucial to control Ptx-induced activation of NF-κB and its downstream responses, since this pathway regulates genes central to metastasis, angiogenesis and drug-resistance.8, 9, 10, 11, 20 The overexpression of P-gp, the efflux pump, driven by Ptx-induced activation of NF-κB has been implicated as a mechanism involved in NF-κB induced drug resistance.21, 22 Several other mechanisms downstream of NF-κB are also indicated in Ptx resistance.23, 24, 25 Therefore, the key objective of our

Acknowledgement

Authors thank Mr. Sajin P. Ravi for SEM analysis, Ms. Sreerekha P. R. for FACS analysis, Mr. Sarath S for Confocal analysis, and Ms. Aswathy J for the technical help extended for immunohistochemistry. Authors also thank Amrita Vishwa Vidyapeetham University for providing all infrastructural support for the research work.

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    Sources of Support for Research: Indian Council of Medical Research, Government of India.

    Conflict of Interest: The authors declare that there are no conflicts of interest.

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