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Morphologically transformable peptide nanocarriers coloaded with doxorubicin and curcumin inhibit the growth and metastasis of hepatocellular carcinoma

Liu, Yun, Liu, Yunxia, Sun, Xinyu, Wang, Yue, Du, Changqing, Bai, Jingkun·2024·Materials Today Bio

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Graphical abstract Graphical abstract Novel NPs of DC/Pep1 for Cancer Therapy. Pep1 encapsulated with DOX and CUR can self-assemble to form NPs (DC/Pep1), which can be targeted to the tumor vasculature and enter the tumor microenvironment in the presence of CREKA. DC/Pep1 undergoes morphological transformations in the acidic tumor microenvironment, resulting in drug release and prolonging drug retention in the tumor tissues in vivo. DC/Pep1 inhibits angiogenesis, promotes DC/Pep1 inhibition of angiogenesis, promotes apoptosis, and inhibits C–C motif chemokine 2 (CCL-2), resulting in effective inhibition of lung metastasis and synergistic antitumor effects.
Graphical abstract Graphical abstract Novel NPs of DC/Pep1 for Cancer Therapy. Pep1 encapsulated with DOX and CUR can self-assemble to form NPs (DC/Pep1), which can be targeted to the tumor vasculature and enter the tumor microenvironment in the presence of CREKA. DC/Pep1 undergoes morphological transformations in the acidic tumor microenvironment, resulting in drug release and prolonging drug retention in the tumor tissues in vivo. DC/Pep1 inhibits angiogenesis, promotes DC/Pep1 inhibition of angiogenesis, promotes apoptosis, and inhibits C–C motif chemokine 2 (CCL-2), resulting in effective inhibition of lung metastasis and synergistic antitumor effects.CC BY-NC-ND · PubMed Central
Fig. 1 Novel NPs of DC/Pep1 for Cancer Therapy. Pep1 encapsulated with DOX and CUR can self-assemble to form NPs (DC/Pep1), which can be targeted to the tumor vasculature and enter the tumor microenvironment in the presence of CREKA. DC/Pep1 undergoes morphological transformations in the acidic tumor microenvironment, resulting in drug release and prolonging drug retention in the tumor tissues in vivo. DC/Pep1 inhibits angiogenesis, promotes apoptosis, and inhibits C–C motif chemokine ligand-2 (CCL-2), resulting in effective inhibition of lung metastasis and synergistic antitumor effects (by Figdraw).
Fig. 1 Novel NPs of DC/Pep1 for Cancer Therapy. Pep1 encapsulated with DOX and CUR can self-assemble to form NPs (DC/Pep1), which can be targeted to the tumor vasculature and enter the tumor microenvironment in the presence of CREKA. DC/Pep1 undergoes morphological transformations in the acidic tumor microenvironment, resulting in drug release and prolonging drug retention in the tumor tissues in vivo. DC/Pep1 inhibits angiogenesis, promotes apoptosis, and inhibits C–C motif chemokine ligand-2 (CCL-2), resulting in effective inhibition of lung metastasis and synergistic antitumor effects (by Figdraw).CC BY-NC-ND · PubMed Central
Fig. 2 Characterization and Cytotoxicity Assay of Peptides. (a) Structures of Pep1 and Pep2. (b) TEM images of Pep1, DC/Pep1, DC/Pep1 (pH 5), Pep2, DC/Pep2, and DC/Pep2 (pH 5). Scale bar: 100 nm. (c) Viability of HepG2 cells and HUVECs after 48 h of peptide treatment. **P < 0.01; ***P < 0.001. (d) Changes in the zeta potentials of Pep1, DC/Pep1 and DC/Pep1 at pH 5 after 12 h and 120 h and the changes in Pep2 potential after the same treatments. (e) Cumulative DOX and CUR release profiles from Pep1 before and after the addition of acid.
Fig. 2 Characterization and Cytotoxicity Assay of Peptides. (a) Structures of Pep1 and Pep2. (b) TEM images of Pep1, DC/Pep1, DC/Pep1 (pH 5), Pep2, DC/Pep2, and DC/Pep2 (pH 5). Scale bar: 100 nm. (c) Viability of HepG2 cells and HUVECs after 48 h of peptide treatment. **P < 0.01; ***P < 0.001. (d) Changes in the zeta potentials of Pep1, DC/Pep1 and DC/Pep1 at pH 5 after 12 h and 120 h and the changes in Pep2 potential after the same treatments. (e) Cumulative DOX and CUR release profiles from Pep1 before and after the addition of acid.CC BY-NC-ND · PubMed Central
Fig. 3 Effects of Pep1 on Tumor Cell Uptake, Retention and Migration. CLSM images of HepG2 cells coexisting with DC, DC/Pep2, and DC/Pep1 for (a) 0.25 h and (b) 0.5 h. Scale bar: 50 μm. Uptake of DOX and CUR in HepG2 cells after coculture with DC, DC/Pep2, and DC/Pep1 for (c) 0.25 h and (d) 0.5 h was detected by FCM. (e) CLSM images of HepG2 cells cocultivated with DC, DC/Pep2, and DC/Pep1 for 72 h. Scale bar: 50 μm. (f) WB images indicating Vimentin and E-cadherin and (g) quantitative analysis. (h) Quantitative evaluation of HepG2 cells at 24 h and 48 h intervals following the scratch assay. *P < 0.05; **P < 0.01; ***P < 0.001; ns, P > 0.05 relative to DC/Pep1.
Fig. 3 Effects of Pep1 on Tumor Cell Uptake, Retention and Migration. CLSM images of HepG2 cells coexisting with DC, DC/Pep2, and DC/Pep1 for (a) 0.25 h and (b) 0.5 h. Scale bar: 50 μm. Uptake of DOX and CUR in HepG2 cells after coculture with DC, DC/Pep2, and DC/Pep1 for (c) 0.25 h and (d) 0.5 h was detected by FCM. (e) CLSM images of HepG2 cells cocultivated with DC, DC/Pep2, and DC/Pep1 for 72 h. Scale bar: 50 μm. (f) WB images indicating Vimentin and E-cadherin and (g) quantitative analysis. (h) Quantitative evaluation of HepG2 cells at 24 h and 48 h intervals following the scratch assay. *P < 0.05; **P < 0.01; ***P < 0.001; ns, P > 0.05 relative to DC/Pep1.CC BY-NC-ND · PubMed Central
Fig. 4 Effects of Drug-loaded Peptides on HepG2 Cell Apoptosis and Angiogenesis. (a) WB analysis and (b) quantitative analysis of Caspase-3 activation. (c) Flow chart of HepG2 cell apoptosis (Q1: necrotic cells; Q2: late apoptotic cells; Q3: early apoptotic cells; and Q4: live cells) and (d) quantitative analysis. (e) Representative photographs and (f) quantitative analysis of the numbers of tubes formed and the capture of tubular structures in HUVECs. Scale bar: 200 μm. *P < 0.05; **P < 0.01; ***P < 0.001; ns, P > 0.05 relative to DC/Pep1.
Fig. 4 Effects of Drug-loaded Peptides on HepG2 Cell Apoptosis and Angiogenesis. (a) WB analysis and (b) quantitative analysis of Caspase-3 activation. (c) Flow chart of HepG2 cell apoptosis (Q1: necrotic cells; Q2: late apoptotic cells; Q3: early apoptotic cells; and Q4: live cells) and (d) quantitative analysis. (e) Representative photographs and (f) quantitative analysis of the numbers of tubes formed and the capture of tubular structures in HUVECs. Scale bar: 200 μm. *P < 0.05; **P < 0.01; ***P < 0.001; ns, P > 0.05 relative to DC/Pep1.CC BY-NC-ND · PubMed Central
Fig. 5 Distribution of DiR in H22 Tumor-Bearing Mice in Vivo and in Isolated Tumors and Major Organs. (a) Fluorescence images of live mice were observed in vivo following the administration of DiR, DiR/Pep2 and DiR/Pep1 via intravenous injection during specific time intervals. (b) Fluorescence images of tissues ex vivo at the designated time intervals and (c) quantitative analysis. *P < 0.05; **P < 0.01.
Fig. 5 Distribution of DiR in H22 Tumor-Bearing Mice in Vivo and in Isolated Tumors and Major Organs. (a) Fluorescence images of live mice were observed in vivo following the administration of DiR, DiR/Pep2 and DiR/Pep1 via intravenous injection during specific time intervals. (b) Fluorescence images of tissues ex vivo at the designated time intervals and (c) quantitative analysis. *P < 0.05; **P < 0.01.CC BY-NC-ND · PubMed Central
Fig. 6 In Vivo Antitumor Effects. (a) In vivo antitumor assay. (b) Images, (c) weights and (d) volumes of H22 tumors excised on day 14. (e) Rates of tumor inhibition achieved. (f) Differences in the weight of mice with tumors. *P < 0.05; **P < 0.01; ***P < 0.001.
Fig. 6 In Vivo Antitumor Effects. (a) In vivo antitumor assay. (b) Images, (c) weights and (d) volumes of H22 tumors excised on day 14. (e) Rates of tumor inhibition achieved. (f) Differences in the weight of mice with tumors. *P < 0.05; **P < 0.01; ***P < 0.001.CC BY-NC-ND · PubMed Central
Fig. 7 Representative Histological Images of the Main Organs and Tumors of H22 Tumor-Bearing Mice after Different Treatments (Saline, DC, DOX/Pep2, DOX/Pep1, DC/Pep2, DC/Pep1). (a) After various treatments, the heart, liver, spleen, lung, kidney and tumor tissues of mice with tumors were examined using H&E staining. Scale bar: 30 μm. (b) Immunohistological staining for CD31 and (c) TUNEL staining of tumor sections treated with different medications. Scale bar: 30 μm.
Fig. 7 Representative Histological Images of the Main Organs and Tumors of H22 Tumor-Bearing Mice after Different Treatments (Saline, DC, DOX/Pep2, DOX/Pep1, DC/Pep2, DC/Pep1). (a) After various treatments, the heart, liver, spleen, lung, kidney and tumor tissues of mice with tumors were examined using H&E staining. Scale bar: 30 μm. (b) Immunohistological staining for CD31 and (c) TUNEL staining of tumor sections treated with different medications. Scale bar: 30 μm.CC BY-NC-ND · PubMed Central
Fig. 8 Antimetastatic Effects of Different Preparations in the H22 Tumor Model. (a) In vivo antimetastasis assay in tumor-bearing mice. (b) Images of lung metastasis and (c) the count of nodules with lung metastases. **P < 0.01; ***P < 0.001; ns, P > 0.05. (d) Body weight increase patterns of mice throughout the duration of the treatment. (e) H&E-stained lung sections. Scale bar: 50 μm. (f) The levels of CCL-2 in lung sections determined by immunofluorescence. Scale bar: 50 μm.
Fig. 8 Antimetastatic Effects of Different Preparations in the H22 Tumor Model. (a) In vivo antimetastasis assay in tumor-bearing mice. (b) Images of lung metastasis and (c) the count of nodules with lung metastases. **P < 0.01; ***P < 0.001; ns, P > 0.05. (d) Body weight increase patterns of mice throughout the duration of the treatment. (e) H&E-stained lung sections. Scale bar: 50 μm. (f) The levels of CCL-2 in lung sections determined by immunofluorescence. Scale bar: 50 μm.CC BY-NC-ND · PubMed Central

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Authors
Liu, Y., Liu, Y., Sun, X., Wang, Y., Du, C., & Bai, J.
Venue
Materials Today Bio
Publisher
Elsevier BV
Published
2024-02
Volume
24
Pages
100903
ISSN
2590-0064
Cited by
19
References
1
Language
en
Funders
Natural Science Foundation of Shandong Province (ZR2023MC122)

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