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Concurrent Treatment with Anti-DLL4 Enhances Antitumor and Proapoptotic Efficacy of a γ-Secretase Inhibitor in Gastric Cancer  Muxing Kang, Yaoyi Zhang,

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Presentation on theme: "Concurrent Treatment with Anti-DLL4 Enhances Antitumor and Proapoptotic Efficacy of a γ-Secretase Inhibitor in Gastric Cancer  Muxing Kang, Yaoyi Zhang,"— Presentation transcript:

1 Concurrent Treatment with Anti-DLL4 Enhances Antitumor and Proapoptotic Efficacy of a γ-Secretase Inhibitor in Gastric Cancer  Muxing Kang, Yaoyi Zhang, Xiaoli Jin, Guofeng Chen, Yi Huang, Dan Wu, Guogang Li, Jianzhen Shan, Pintong Huang, Jian Chen  Translational Oncology  Volume 11, Issue 3, Pages (June 2018) DOI: /j.tranon Copyright © 2018 The Authors Terms and Conditions

2 Figure 1 DLL4 protein expression in SGC-7901-GFP cells.
Membranous expression pattern of DLL4 protein in SGC-7901 cells detected by the rosette formation test with anti-DLL4 antibody conjugated microbubbles. (A) No rosette formation observed with SGC-7901 cells treated with unconjugated microbubbles. (B) Rosette-like structure formed with SGC-7901 cells treated with anti-DLL4 antibody conjugated microbubbles. (C) Rosette formation blocked by adding anti-DLL4 antibody prior to SGC-7901 cells being treated with anti-DLL4 antibody conjugated microbubbles. Bars = 50 μm. DLL4 expression in SGC-7901 cells was detected by immunofluorescence. (D) Red fluorescence showing DLL4 expression in SGC-7901 cells. (E) DAPI showing SGC-7901 nuclei. (F) Merged image of A and B. Translational Oncology  , DOI: ( /j.tranon ) Copyright © 2018 The Authors Terms and Conditions

3 Figure 2 Effect of anti-DLL4 and DAPT on apoptosis and cell cycle in gastric cancer cells. (A to D) Cells were stained with Annexin V and PI, following which cell apoptosis in each group was detected by flow cytometry. (E to H) Cell cycle phase distribution was measured by staining DNA with PI followed by flow cytometry. (I and J) Cell apoptosis rate and cell cycle distribution in each group were analyzed. *P < .01, when compared with control group. Translational Oncology  , DOI: ( /j.tranon ) Copyright © 2018 The Authors Terms and Conditions

4 Figure 3 Effect of anti-DLL4 and DAPT on cell invasion of gastric cancer cells. Cell invasion was assessed by Transwell invasion assay. (A) Representative Transwell cells images in each group. (B) Comparison of immigrated cell numbers in each group. *P < .01, when compared to control group. #P < .01, when compared to DAPT group. Translational Oncology  , DOI: ( /j.tranon ) Copyright © 2018 The Authors Terms and Conditions

5 Figure 4 Conventional ultrasound imaging and contrast-enhanced ultrasound imaging of orthotopic gastric tumor in mice for UTMD. (A) B-mode imaging of tumor. (B) Color Doppler imaging of blood flow in tumor. (C) CPS mode imaging of microbubble wash-in in tumor. (D) CPS mode imaging of microbubble destruction in tumor by UTMD. Translational Oncology  , DOI: ( /j.tranon ) Copyright © 2018 The Authors Terms and Conditions

6 Figure 5 Effect of anti-DLL4 and DAPT on tumor growth in orthotopic gastric tumor models. Tumor growth was monitored and quantified by real-time whole-body fluorescence imaging. Final tumor weight was determined at autopsy. (A) In vivo whole-body fluorescence imaging of tumor before treatment and 18 days after treatment in each group. (B) Markedly increased necrogenous reaction in the parenchymal of tumor in the combined treatment group was observed by H&E staining. (C) Tumor growth curves in each group. (D) Final tumor weights at the end of the study period in each group. *P < .01, when compared to MB+UTMD control. #P < .01, when compared to MB+DAPT+UTMD group. Translational Oncology  , DOI: ( /j.tranon ) Copyright © 2018 The Authors Terms and Conditions

7 Figure 6 Effect of anti-DLL4 and DAPT on tumor cell apoptosis in in vivo models. The apoptosis of the tumor cells was detected by TUNEL staining. (A) Representative TUNEL images for cell apoptosis in each group (100× magnification). (B) Quantitation of cell apoptosis by AOD. *P < .01, when compared to the MB+UTMD control. #P < .01, when compared to the MB+DAPT+UTMD group. Translational Oncology  , DOI: ( /j.tranon ) Copyright © 2018 The Authors Terms and Conditions

8 Figure 7 Effect of anti-DLL4 and DAPT on the expression of apoptosis-related genes and the body weight of mice in orthotopic gastric tumor models. (A) Representative immunohistochemistry staining for BAX, Bcl-2, and P53 in the tumors of each group. 400× magnification. (B) AOD of immunohistochemistry staining of tumor samples in each group. *P < .05, when compared to the MB+UTMD control. (C) No significant anti-DLL4– and DAPT-related body weight loss was observed in the treated mice. Translational Oncology  , DOI: ( /j.tranon ) Copyright © 2018 The Authors Terms and Conditions

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