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Article type: Research Article
Authors: Li, Mina | Zhu, Yilongb; c | Bai, Bingb | Fang, Jinbob | Yao, Weic; d | Li, Yiquanb; c | Li, Shanzhib | Li, Xiaoa; b; c; * | Jin, Ningyia; b; c; * | Jiang, Rihuaa; *
Affiliations: [a] Department of Dermatology, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China | [b] Academicians Workstation of Jilin Province, Changchun University of Chinese Medicine, Changchun, Jilin, China | [c] Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, Jilin, China | [d] Center for Disease Control and Prevention, Agency for Offices Administration, Central Military Commission, Beijing, China
Correspondence: [*] Corresponding authors: Xiao Li, Department of Dermatology, China-Japan Union Hospital of Jilin University, XianTai Da Street No. 126, Jilin 130122, China. Tel.: +86 431 86985929; E-mail: skylee6226@163.com. Ningyi Jin, Department of Dermatology, China-Japan Union Hospital of Jilin University, XianTai Da Street No. 126, Jilin 130122, China. Tel.: +86 431 86985929; E-mail: lixiao06@mails.jlu.edu.cn. Rihua Jiang, Department of Dermatology, China-Japan Union Hospital of Jilin University, XianTai Da Street No. 126, Jilin 130122, China. Tel.: +86 431 86985929; E-mail: jiangrh@jlu.edu.cn.
Abstract: BACKGROUND: To explore the suppressive effect of Apoptin-loaded oncolytic adenovirus (Ad-VT) on luciferase-labeled human melanoma cells in vitro and in vivo. METHODS: The stable luciferase-expressing human melanoma cells A375-luc or M14-luc were obtained by transfecting the plasmid pGL4.51 and selection with G418, followed by luciferase activity, genetic stability and bioluminescence intensity assays. In vitro, the inhibitory effects of Ad-VT on A375-luc or M14-luc were evaluated using the MTS cell proliferation, FITC-Annexin V apoptosis detection, transwell migration, Matrigel invasion and scratch assays. The inhibition pathway in Ad-VT-infected A375-luc and M14-luc cells were analyzed by JC-1 staining and Western-blot detection of mitochondrial apoptosis-related proteins. In vivo, the suppressive effects of Ad-VT on A375-luc or M14-luc were assessed by living imaging technology, tumor volume, bioluminescence intensity, survival curves and immunohistochemical analysis of the tumors from the xenograft tumor model BALB/c nude mice. RESULTS: The growth and migration of A375-luc and M14-luc were significantly inhibited by Ad-VT in vitro. The evaluations of A375-luc and M14-luc tumor models in BALB/c nude mice were successfully performed using living imaging technology. Ad-VT had an anti-tumor effect by reducing tumor growth and increasing survival in vivo. Ad-VT significantly changed the mitochondrial membrane potential by triggering the the mitochondrial release of apoptosis-related proteins, AIF (apoptosis inducing factor), ARTS (Apoptosis-Related Proteins), and Cyto-c (cytochrome c) from the mitochondria. CONCLUSION: Ad-VT reduced the mitochondrial membrane potential in A375-luc or M14-luc cells and induced the mitochondrial release of AIF, ARTS and Cyto-C. Ad-VT induced apoptosis in A375-luc or M14-luc cells via the mitochondrial apoptotic pathway.
Keywords: Luciferase labeled A375 or M14, living imaging technology, oncolytic adenovirus, tumor suppression, BALB/C nude mice model
DOI: 10.3233/CBM-203150
Journal: Cancer Biomarkers, vol. 32, no. 3, pp. 251-262, 2021
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