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Open AccessArticle

Delivery of Protein Kinase A by CRISPRMAX and Its Effects on Breast Cancer Stem-Like Properties

by 1,2,3,4,5,†, 3,6,†, 3,4,5, 3,4,5, 7, 3,4,5,8,*, 4,5,* and 3,4,5,*
1
Experimental Hematology and Biochemistry Lab, Beijing Institute of Radiation Medicine, Beijing 100850, China
2
Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing 100850, China
3
Pharmaceutical Sciences Laboratory, Åbo Akademi University, 20520 Turku, Finland
4
Turku Bioscience Center, University of Turku, 20520 Turku, Finland
5
Turku Bioscience Center, Åbo Akademi University, 20520 Turku, Finland
6
Institute of Biomedicine, Faculty of Medicine, University of Turku, 20520 Turku, Finland
7
Department of Radiology, Affiliated Hospital of Jiangsu University, Jiangsu University, Zhenjiang 212001, China
8
Department of Endocrinology, Key Laboratory of National Health and Family Planning Commission for Male Reproductive Health, National Research Institute for Family Planning, Beijing 100081, China
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this paper.
Pharmaceutics 2021, 13(1), 11; https://doi.org/10.3390/pharmaceutics13010011
Received: 6 November 2020 / Revised: 14 December 2020 / Accepted: 18 December 2020 / Published: 23 December 2020
(This article belongs to the Special Issue Hybrid Multifunctional Drug Delivery Systems)
Protein kinase A (PKA) activation has recently been reported to inhibit epithelial-mesenchymal transition (EMT) and cancer stem cell (CSC) ability, which is considered to be responsible for chemoresistance and tumor recurrence in patients. While current studies mainly focus on gene manipulation of the EMT process, the direct delivery of PKA enzymes to cancer cells has never been investigated. Here, we utilize the commercial Lipofectamine CRISPRMAX reagent to directly deliver PKAs to breast cancer cells and evaluate its effects on EMT regulation. We optimized the delivery parameters with fluorescent-labeled bovine serum albumin, and successfully delivered fluorescent PKAs through CRISPRMAX into breast cancer cells. Then, we evaluated the biological effects by immunofluorescence, flow cytometry, mammosphere assay, and chemoresistance assay. Our data showed the expression of EMT-related markers, α-smooth muscle actin and N-cadherin, was downregulated after CRISPRMAX-PKA treatment. Although the CD44+/CD24 population did not change considerably, the size of mammospheres significantly decreased. In paclitaxel and doxorubicin chemoresistance assays, we noticed PKA delivery significantly inhibited paclitaxel resistance rather than doxorubicin resistance. Taken together, these results suggest our direct enzyme delivery can be a potential strategy for inhibiting EMT/CSC-associated traits, providing a safer approach and having more clinical translational efficacy than gene manipulation. This strategy will also facilitate the direct testing of other target enzymes/proteins on their biological functions. View Full-Text
Keywords: CRISPRMAX; drug delivery; protein kinase A; chemoresistance; cancer stem cells; epithelial-mesenchymal transition CRISPRMAX; drug delivery; protein kinase A; chemoresistance; cancer stem cells; epithelial-mesenchymal transition
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MDPI and ACS Style

Zhou, J.-N.; Rautio, T.-C.; Liu, C.; Xu, X.-Y.; Wang, D.-Q.; Guo, Y.; Eriksson, J.; Zhang, H. Delivery of Protein Kinase A by CRISPRMAX and Its Effects on Breast Cancer Stem-Like Properties. Pharmaceutics 2021, 13, 11. https://doi.org/10.3390/pharmaceutics13010011

AMA Style

Zhou J-N, Rautio T-C, Liu C, Xu X-Y, Wang D-Q, Guo Y, Eriksson J, Zhang H. Delivery of Protein Kinase A by CRISPRMAX and Its Effects on Breast Cancer Stem-Like Properties. Pharmaceutics. 2021; 13(1):11. https://doi.org/10.3390/pharmaceutics13010011

Chicago/Turabian Style

Zhou, Jun-Nian; Rautio, Tzu-Chen; Liu, Chang; Xu, Xiao-Yu; Wang, Dong-Qing; Guo, Yong; Eriksson, John; Zhang, Hongbo. 2021. "Delivery of Protein Kinase A by CRISPRMAX and Its Effects on Breast Cancer Stem-Like Properties" Pharmaceutics 13, no. 1: 11. https://doi.org/10.3390/pharmaceutics13010011

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