Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (34): 5468-5472.doi: 10.3969/j.issn.2095-4344.1968

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Construction of a gene-loaded multifunctional polymer microbubble targeting breast cancer cells

Lin Min1,2, Wei Weili1, He Yimi1, Chen Zhikui1
  

  1. 1Department of Ultrasound, Affiliated Union Hospital of Fujian Medical University, Fuzhou 350001, Fujian Province, China; 2Longyan First Hospital of Fujian Medical University, Longyan 364000, Fujian Province, China
  • Received:2019-06-26 Online:2019-12-08 Published:2019-12-08
  • Contact: Chen Zhikui, MD, Associate chief physician, Associate professor, Department of Ultrasound, Affiliated Union Hospital of Fujian Medical University, Fuzhou 350001, Fujian Province, China
  • About author:Lin Min, Master, Attending physician, Department of Ultrasound, Affiliated Union Hospital of Fujian Medical University, Fuzhou 350001, Fujian Province, China; Longyan First Hospital of Fujian Medical University, Longyan 364000, Fujian Province, China
  • Supported by:

    The Natural Science Foundation of Fujian Province (General Program), No. 2014J01416 (to CZK)

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Abstract:

BACKGROUND: Polymer microbubbles have good stability and long retention time.
OBJECTIVE: To construct gene-loaded polymer microbubbles and to assess their ability to target receptors in vitro.
METHODS: The microbubbles were prepared with polymers (mPEG-PLLA, PLGA-PEG-COOH and DC-Chol) as shell and PFP as core by emulsion method, then modified with herceptin and cationic polymer coated plasmid DNA to form the gene-loaded targeting polymer microbubbles. The morphology, size, and distribution were determined by optical microscopy. The rate of the polymer microbubbles conjugating with both plasmid DNA and trastuzumab-IgG-FITC was determined by fluorescence microscopy and flow cytometry. When human epidermal growth factor receptor  2 (+) breast cancer cells grew to about 80% confluency, they were divided into two groups: one group was incubated with polymer microbubbles for 1 hour; the other group was pre-incubated with trastuzumab for 1 hour, and then with polymer microbubbles for another 1 hour. The ability of the microbubbles binding to human epidermal growth factor receptor 2 (+) breast cancer cells in vitro was detected under a microscope.
RESULTS AND CONCLUSION: The gene-targeted polymer microbubbles were round and well dispersed, with an average particle size of (3.0±1.5) μm and they had uniform size distribution with a concentration of 8.8×1010/L. Under the fluorescence microscope, trastuzumab-IgG-FITC on the surface of gene-loaded breast cancer cells-targeting polymer microbubbles emitted green fluorescence, and plasmid DNA-PI emitted red fluorescence. Flow cytometry showed that the rate of polymer microbubbles both conjugating with plasmid DNA and trastuzumab was 96.28%. After the gene-loaded polymer microbubbles and breast cancer cells were co-incubated and for 1 hour, microscopy showed that a large number of polymer microbubbles were bound on the cell surface, and after blocking with trastuzumab, the amount of polymer microbubbles bounding to breast cancer cells was significantly reduced. The results showed that the polymer microbubbles prepared by the single emulsion method can effectively carry plasmid DNA and antibody drugs, and have good targeting function.

Key words: microbubbles, polymer, single emulsion method, breast cancer, human epidermal growth factor receptor, gene therapy, targeted therapy, flow cytometry

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