Volume 26, Issue 3, Pages 774-783 (March 2018) Exosomes Serve as Nanoparticles to Deliver Anti-miR-214 to Reverse Chemoresistance to Cisplatin in Gastric Cancer  Xinyi Wang, Haiyang Zhang, Ming Bai, Tao Ning, Shaohua Ge, Ting Deng, Rui Liu, Le Zhang, Guoguang Ying, Yi Ba  Molecular Therapy  Volume 26, Issue 3, Pages 774-783 (March 2018) DOI: 10.1016/j.ymthe.2018.01.001 Copyright © 2018 The American Society of Gene and Cell Therapy Terms and Conditions

Figure 1 Expression of miR-214 and Its Potential Targets (A) Expression levels of miR-214 in ges-1, SGC7901, and SGC7901/DDP were examined by qRT-PCR. (B) Potential targets of miR-214 were predicted via LC-ESI-MS/MS in SGC7901. Different shades represented different expression levels of relative genes. The darker it was, the higher the gene expressed. (C) Western blots were conducted to verify the possible targets in both SGC7901 and SGC7901/DDP cells. (D) Gray analysis of (C). (E) Cell viability for different drug concentrations in SGC7901 was tested by CCK-8, and the inhibition ratio was calculated. (F) Inhibition ratio in SGC7901/DDP cells. *p < 0.05; **p < 0.01; ***p < 0.001. All error bars stand for SE. Molecular Therapy 2018 26, 774-783DOI: (10.1016/j.ymthe.2018.01.001) Copyright © 2018 The American Society of Gene and Cell Therapy Terms and Conditions

Figure 2 Exo-anti-214 Fuses into Cells and Regulates Potential Targets (A) Image of exosomes from HEK293T cell medium through a transmission electron microscope. (B) Marker proteins for exosomes were determined via western blot and assessed by gray analysis. (C) Exosomes stained with PKH26 fused into gastric cancer cells. (D) qRT-PCR was applied to test the expression levels of miR-214 and anti-miR-214 in HEK exosomes. (E) Expression levels of the potential target proteins in SGC7901 and SGC7901/DDP cells were detected by western blot. (F) Gray analysis of (E). *p < 0.05; **p < 0.01; ***p < 0.001. All error bars stand for SE. Molecular Therapy 2018 26, 774-783DOI: (10.1016/j.ymthe.2018.01.001) Copyright © 2018 The American Society of Gene and Cell Therapy Terms and Conditions

Figure 3 Exo-anti-214 Reverses the Resistance to DDP in Gastric Cancer Cells (A) A CCK-8 assay was conducted to determine the cell viability in SGC7901. (B) Cell viability in SGC7901/DDP cells. (C) Cell apoptosis was tested through flow cytometry in SGC7901 and SGC7901/DDP cells. (D) Quantitative analysis of (C). *p < 0.05; **p < 0.01; ***p < 0.001. All error bars stand for SE. Molecular Therapy 2018 26, 774-783DOI: (10.1016/j.ymthe.2018.01.001) Copyright © 2018 The American Society of Gene and Cell Therapy Terms and Conditions

Figure 4 Exo-anti-214 Enhances the Cytotoxicity of DDP in Gastric Cancer Cells (A) A transwell assay was performed to examine the cell migration ability in both cell lines. (B) Quantitative analysis of (A). (C) H2AFX was stained via immunofluorescence to detect DNA damage. (D) Quantitative analysis of (C). **p < 0.01. All error bars stand for SE. Molecular Therapy 2018 26, 774-783DOI: (10.1016/j.ymthe.2018.01.001) Copyright © 2018 The American Society of Gene and Cell Therapy Terms and Conditions

Figure 5 Systemically Injected Exo-anti-214 Sensitizes the Response to DDP In Vivo (A) A flow chart depicting the in vivo experimental design. (B) Alterations of tumor volume in the treatment and two control groups. (C) Images of tumors in mice (n = 15). (D) Weight measurements of the tumors. (E) Levels of anti-miR-214 in serum exosomes were detected by qRT-PCR. (F) Expression levels of miR-214 in serum exosomes. (G) Expression levels of miR-214 in tumors. (H) Potential target proteins were analyzed by western blot. (I) Gray analysis of (H). *p < 0.05; **p < 0.01; ***p < 0.001. All error bars stand for SE. Molecular Therapy 2018 26, 774-783DOI: (10.1016/j.ymthe.2018.01.001) Copyright © 2018 The American Society of Gene and Cell Therapy Terms and Conditions