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Downregulation of CD73 in 4T1 breast cancer cells through siRNA-loaded chitosan-lactate nanoparticles

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Tumor Biology

Abstract

The immunosuppressive factors in tumor microenvironment enhance tumor growth and suppress anti-tumor immune responses. Adenosine is an important immunosuppressive factor which can be secreted by both tumor and immune cells trough action of two cell surface ecto-nucleotidase molecules CD39 and CD73. Blocking the adenosine generating molecules has emerged as an effective immunotherapeutic approach for treatment of cancer. In this study, CD73-siRNA encapsulated into chitosan-lactate (ChLa) nanoparticles (NPs) was employed to suppress the expression of CD73 molecule on 4T1 breast tumor cells, in vitro. ChLa NPs were generated through ionic gelation of ChLa by tripolyphosphate (TPP). Small interfering RNA (SiRNA)-loaded NPs had about 100 nm size with a polydispersive index below 0.3 and a zeta potential about 13. Our results showed that ChLa NPs with Ch 50 kDa exhibit the best physicochemical features with the high siRNA encapsulation capacity. Synthesized NPs were able to fully bind with siRNA, protect them against serum and heparin degradation, and promote the transfection process. While the NPs exhibited low toxicity during 72 h cell culture, the transfection of Ch-plasmid expressing green fluorescent protein (pEGFP) NPs was efficient in 4T1 cells with a transfection rate of 53.6 % as detected by flow cytometry. In addition, CD73-siRNA-loaded ChLa NPs could efficiently suppress the expression of CD73 as assayed by real-time polymerase chain reaction and flow cytometry. As a conclusion, CD73-siRNA-loaded ChLa NPs may be considered as a promising therapeutic tool for cancer therapy; however, further in vivo investigations are necessary.

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Acknowledgments

We would like to thank, Maryam Ajami, Zahra Mirzaie, and Reyhaneh Varshochian for their excellent technical support. This study was supported in part by a grant from Tehran University of Medical Sciences (grant number: 24869).

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Authors and Affiliations

  1. Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

    Farhad Jadidi-Niaragh, Alireza Razavi, Hadi Hassannia & Fazel Shokri

  2. Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

    Farhad Jadidi-Niaragh & Mehdi Yousefi

  3. Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

    Farhad Jadidi-Niaragh & Mehdi Yousefi

  4. Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, 1714614411, Iran

    Fatemeh Atyabi, Ali Rastegari & Melika Kiani

  5. Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran

    Fatemeh Atyabi

  6. Food and Drug Control Laboratories and Food and Drug Laboratory Research Centre, MOHME, Tehran, Iran

    Esmail Mollarazi

  7. Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

    Nasim Kheshtchin & Jamshid Hadjati

  8. Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran

    Fazel Shokri

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  1. Farhad Jadidi-Niaragh
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Correspondence to Fatemeh Atyabi or Fazel Shokri.

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Jadidi-Niaragh, F., Atyabi, F., Rastegari, A. et al. Downregulation of CD73 in 4T1 breast cancer cells through siRNA-loaded chitosan-lactate nanoparticles. Tumor Biol. 37, 8403–8412 (2016). https://doi.org/10.1007/s13277-015-4732-0

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  • DOI: https://doi.org/10.1007/s13277-015-4732-0

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