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Molecular Therapy - Nucleic Acids

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Presentation on theme: "Molecular Therapy - Nucleic Acids"— Presentation transcript:

1 Molecular Therapy - Nucleic Acids
MiR-375 and Doxorubicin Co-delivered by Liposomes for Combination Therapy of Hepatocellular Carcinoma  Yin-Ping Fan, Jia-Zhi Liao, Ya-Qi Lu, De-An Tian, Feng Ye, Peng-Xuan Zhao, Guang-Ya Xiang, Wang-Xian Tang, Xing-Xing He  Molecular Therapy - Nucleic Acids  Volume 7, Pages (June 2017) DOI: /j.omtn Copyright © 2017 The Author(s) Terms and Conditions

2 Figure 1 Schematic Description of L-miR-375/DOX-NPs for Synergetic Anti-tumor and Reversal of Drug-Resistance Mechanism in HCC After swallowed by HCC cells, L-miR-375/DOX-NPs escape from the endosome, doxorubicin enter into the nucleus, and give full play to cytotoxic effect, while miR-375 is released to cytoplasm and works on downstream genes, suppressing the malignant characteristics of HCC cells. The combination of doxorubicin and miR-375 show enhanced anti-tumor efficiency and partial reversal of drug resistance. Molecular Therapy - Nucleic Acids 2017 7, DOI: ( /j.omtn ) Copyright © 2017 The Author(s) Terms and Conditions

3 Figure 2 Uptake of L-miR-375/DOX-NPs in SMMC-7721 and HepG2 Cells
(A) Cells after incubated with free DOX, L-DOX-NPs, and L-miR-375/DOX-NPs at 37°C for 1 hr and then were observed under a fluorescence microscope. (B) Flow cytometry indicated the uptake rate of MiR-375 and doxorubicin co-delivered by liposomes in SMMC-7721 and HepG2 cells. Molecular Therapy - Nucleic Acids 2017 7, DOI: ( /j.omtn ) Copyright © 2017 The Author(s) Terms and Conditions

4 Figure 3 L-miR-375/DOX-NPs Upregulation of miR-375 Efficiently and the Anti-tumor Activities of L-miR-375/DOX-NPs In Vitro (A) The release of mature miR-375 after treated with L-miR-375/DOX-NPs for 48 hr. (B) Viabilities of SMMC-7721and HepG2 cells treated with free DOX, L-DOX-NPs, L-miR-375-NPs, and L-miR-375/DOX-NPs were measured by CCK-8 assay at 24 hr, 48 hr, and 72 hr, respectively. (C and D) Migration and invasion abilities of SMMC-7721 cells treated with PBS, free DOX, L-DOX-NPs, L-miR-375-NPs, and L-miR-375/DOX-NPs were determined by Transwell and wound healing assays. (E) Cell apoptosis were analyzed by apoptosis kit, cell nucleus were stained with Hoechst (F) Cell cycles were analyzed by flow cytometry. The data are expressed as (mean ± SEM, n = 3, *p < 0.05; **p < 0.01; ***p < 0.001; and ****p < ). Molecular Therapy - Nucleic Acids 2017 7, DOI: ( /j.omtn ) Copyright © 2017 The Author(s) Terms and Conditions

5 Figure 4 L-miR-375/DOX-NPs Exerts Anti-tumor Effect by Simultaneously Acting on the Target Gene of miR-375 and Doxorubicin (A and B) The protein expressions of AEG-1, YAP1, and ATG7 were detected by western blot after treatment with L-miR-NC-NPs, L-miR-375-NPs, and L-miR-375/DOX-NPs for 72 hr in SMMC-7721 and HepG2 cells. (C and D) The protein levels of Bax, Bcl-2, p-JNK, p-p38, and Caspase-3 were detected by western blot after treated with DOX, L-DOX-NPs, and L-miR-375/DOX-NPs for 72 hr. GAPDH was used as a loading control. All of the data have been repeated at least three times. Molecular Therapy - Nucleic Acids 2017 7, DOI: ( /j.omtn ) Copyright © 2017 The Author(s) Terms and Conditions

6 Figure 5 The Anti-tumor Efficiency of L-miR-375/DOX-NPs In Vivo
(A) Tumors were removed at the end of the experiment (day 14 after the drug administered). (B and C) Growth curves of mouse model and the volume changes of xenograft tumors treated with PBS, free DOX, L-DOX-NPs, L-miR-375-NPs, and L-miR-375/DOX- NPs by intratumoral injection (1 nmol miR-375 per mouse, 1 mg/kg DOX). (D and E) Acute phase changes of liver and renal function after 3 days of drug injection. (F) Hematoxylin and eosin staining analysis of tissues including heart, liver, spleen, lung, and kidney; (B) tumor growth curve (p < 0.05); and (C) tumor volume curve (p < 0.05). Molecular Therapy - Nucleic Acids 2017 7, DOI: ( /j.omtn ) Copyright © 2017 The Author(s) Terms and Conditions

7 Figure 6 L-miR-375/DOX-NPs Reversing Drug Resistance of Doxorubicin in SMMC-7721/ADM Cells (A and B) Identification of SMMC-7721/ADM cells. (C) The comparison of fluorescence intensity about DOX in SMMC-7721/ADM cells after treated with free DOX, L-DOX-NPs, and L-miR-375/DOX-NPs, respectively, at 1 hr and 20 hr. (D) The release of mature miR-375 of SMMC-7721/ADM cells after treated with L-miR-375/DOX-NPs for 48 hr. (E) Molecular mechanism of L-miR-375/DOX-NPs reversing drug resistance in SMMC-7721/ADM cells. (F) The proliferation inhibitory effect of SMMC-7721/ADM cells after treated with free DOX, L-DOX-NPs, and L-miR-375/DOX-NPs with different concentration gradient of DOX. (G) Migration ability of SMMC-7721/ADM cells was determined by Transwell assay. Molecular Therapy - Nucleic Acids 2017 7, DOI: ( /j.omtn ) Copyright © 2017 The Author(s) Terms and Conditions

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