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Original Research

Second-generation aptamer-conjugated PSMA-targeted delivery system for prostate cancer therapy

Xin Wu1 Department of Pharmaceutics, Changhai Hospital, Second Military Medical University, Shanghai

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Baoyue Ding1 Department of Pharmaceutics, Changhai Hospital, Second Military Medical University, Shanghai;2 Medical College of Jiaxing University, Jiaxing

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Jing Gao3 School of Pharmacy, Second Military Medical University, Shanghai

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Huanyun Wang4 School of Pharmacy, Inner Mongolia Medical College, Hohhot

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Wei Fan1 Department of Pharmaceutics, Changhai Hospital, Second Military Medical University, Shanghai

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Xiang Wang1 Department of Pharmaceutics, Changhai Hospital, Second Military Medical University, Shanghai;5 No. 98 Hospital of PLA, Huzhou, People’s Republic of China

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Wei Zhang1 Department of Pharmaceutics, Changhai Hospital, Second Military Medical University, Shanghai

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Xiaoyu Wang1 Department of Pharmaceutics, Changhai Hospital, Second Military Medical University, Shanghai

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Lihua Ye1 Department of Pharmaceutics, Changhai Hospital, Second Military Medical University, Shanghai

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Min Zhang1 Department of Pharmaceutics, Changhai Hospital, Second Military Medical University, Shanghai

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Xueying Ding3 School of Pharmacy, Second Military Medical University, Shanghai

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Jiyong Liu1 Department of Pharmaceutics, Changhai Hospital, Second Military Medical University, Shanghai

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Quangang Zhu1 Department of Pharmaceutics, Changhai Hospital, Second Military Medical University, ShanghaiCorrespondence[email protected] [email protected]

&

Shen Gao1 Department of Pharmaceutics, Changhai Hospital, Second Military Medical University, ShanghaiCorrespondence[email protected] [email protected]

Figures & data

Figure 1 NMR spectra of (A) PAMAM, (B) PAMAM-PEG, and (C) PAMAM-PEG-APT in D₂O at 300 mHz.

Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Figure 1 NMR spectra of (A) PAMAM, (B) PAMAM-PEG, and (C) PAMAM-PEG-APT in D2O at 300 mHz.Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Figure 2 (A) Fluorescence microscopy images (scale bar 50 μm) and (B) fluorescent-activated cell sorting analysis (n = 3, error bars represent a standard deviation) after a 60-minute incubation of DyLight-633-labeled PAMAM-PEG-APT as a function of concentration range against PC3 and LNCaP cells, respectively.

Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Figure 2 (A) Fluorescence microscopy images (scale bar 50 μm) and (B) fluorescent-activated cell sorting analysis (n = 3, error bars represent a standard deviation) after a 60-minute incubation of DyLight-633-labeled PAMAM-PEG-APT as a function of concentration range against PC3 and LNCaP cells, respectively.Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Figure 3 Sizes and zeta potentials of pEGFP/PAMAM-PEG-APT complexes at various N/P ratios.

Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Figure 3 Sizes and zeta potentials of pEGFP/PAMAM-PEG-APT complexes at various N/P ratios.Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Figure 4 (A) Fluorescence microscopy images (scale bar 50 μm) and (B) luciferase activity analysis (n = 4, error bars represent the standard deviation) after transfection of pGL-3/PAMAM-PEG or pGL-3/PAMAM-PEG-APT as a function of N/P ratios against PC3 and LNCaP cells, respectively. *P < 0.05.

Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Figure 4 (A) Fluorescence microscopy images (scale bar 50 μm) and (B) luciferase activity analysis (n = 4, error bars represent the standard deviation) after transfection of pGL-3/PAMAM-PEG or pGL-3/PAMAM-PEG-APT as a function of N/P ratios against PC3 and LNCaP cells, respectively. *P < 0.05.Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Figure 5 Bcl-2, Cyclin D1, and Wnt3a knock down following miRNA/PAMAM-PEG-APT complexes delivery to LNCaP cells. The cells were treated with miRNA/PAMAM-PEG-APT (miRNA) and NC-miRNA/PAMAM-PEG-APT (NC-miRNA) complexes and processed for immunoblotting with anti-Bcl-2, anti-Cyclin D1, and anti-Wnt3a antibodies at 24, 48, and 72 hours post-transfection. β-actin was a loading control.

Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Figure 5 Bcl-2, Cyclin D1, and Wnt3a knock down following miRNA/PAMAM-PEG-APT complexes delivery to LNCaP cells. The cells were treated with miRNA/PAMAM-PEG-APT (miRNA) and NC-miRNA/PAMAM-PEG-APT (NC-miRNA) complexes and processed for immunoblotting with anti-Bcl-2, anti-Cyclin D1, and anti-Wnt3a antibodies at 24, 48, and 72 hours post-transfection. β-actin was a loading control.Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Figure 6 Cell viabilities of LNCaP cells treated with miRNA/PAMAM-PEG-APT, NC-miRNA/PAMAM-PEG-APT, miRNA/PAMAM-PEG complexes, respectively, (n = 3, error bars represent the standard deviation).

Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Figure 6 Cell viabilities of LNCaP cells treated with miRNA/PAMAM-PEG-APT, NC-miRNA/PAMAM-PEG-APT, miRNA/PAMAM-PEG complexes, respectively, (n = 3, error bars represent the standard deviation).Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.