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Using Dual Fluorescence Reporting Genes to Establish an In Vivo Imaging Model of Orthotopic Lung Adenocarcinoma in Mice

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Abstract

Purpose

Lung adenocarcinoma is characterized by a poor prognosis and high mortality worldwide. In this study, we purposed to use the live imaging techniques and a reporter gene that generates highly penetrative near-infrared (NIR) fluorescence to establish a preclinical animal model that allows in vivo monitoring of lung cancer development and provides a non-invasive tool for the research on lung cancer pathogenesis and therapeutic efficacy.

Procedures

A human lung adenocarcinoma cell line (A549), which stably expressed the dual fluorescence reporting gene (pCAG-iRFP-2A-Venus), was used to generate subcutaneous or orthotopic lung cancer in nude mice. Cancer development was evaluated by live imaging via the NIR fluorescent signals from iRFP, and the signals were verified ex vivo by the green fluorescence of Venus from the gross lung. The tumor-bearing mice received miR-16 nucleic acid therapy by intranasal administration to demonstrate therapeutic efficacy in this live imaging system.

Results

For the subcutaneous xenografts, the detection of iRFP fluorescent signals revealed delicate changes occurring during tumor growth that are not distinguishable by conventional methods of tumor measurement. For the orthotopic xenografts, the positive correlation between the in vivo iRFP signal from mice chests and the ex vivo green fluorescent signal from gross lung tumors and the results of the suppressed tumorigenesis by miR-16 treatment indicated that lung tumor size can be accurately quantified by the emission of NIR fluorescence. In addition, orthotopic lung tumor localization can be accurately visualized using iRFP fluorescence tomography in vivo, thus revealing the trafficking of lung tumor cells.

Conclusions

We introduced a novel dual fluorescence lung cancer model that provides a non-invasive option for preclinical research via the use of NIR fluorescence in live imaging of lung.

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Acknowledgments

This research was supported in part by grants MOST-104-2313-B-005-043-MY3 from the Ministry of Science and Technology, ATU-105-S0508 from the Ministry of Education, Taiwan, under the Aiming for Top University plan, and TCVGH-NCHU1047610 from Rong-Hsing cooperation project. The authors would like to thank Dr. Hong-Lin Su for providing the pCAG-Neo-2A-Venus plasmid and Dr. Vladislav Verkhusha for the piRFP plasmid. We thank our colleagues (Drs. Tung-Chou Tsai and Zi-Lun Lai) in the Molecular Embryology and DNA Methylation Laboratory for their help with discussions and technical issues. And we also thank Miss Theresa Röhrig for critically reading and editing the manuscript.

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Authors and Affiliations

  1. Department of Life Sciences, National Chung Hsing University, Taichung, 402, Taiwan

    Cheng-Wei Lai, Chih-Ching Yen, Jiun-Long Wang & Chuan-Mu Chen

  2. Department of Bioresources, Da-Yeh University, Changhua, 515, Taiwan

    Hsiao-Ling Chen

  3. Department of Internal Medicine, China Medical University Hospital, Taichung, 404, Taiwan

    Chih-Ching Yen

  4. Division of Chest Medicine, Department of Internal Medicine, Taichung Veterans General Hospital, Taichung, 407, Taiwan

    Jiun-Long Wang

  5. Department of Physiology, and Institute of Basic Medical Sciences, National Cheng Kung University, Tainan, 701, Taiwan

    Shang-Hsun Yang

  6. Rong-Hsing Translational Medicine Center, iEGG Center, National Chung Hsing University, Taichung, 402, Taiwan

    Chuan-Mu Chen

  7. Agricultural Biotechnology Center, National Chung Hsing University, No. 250, Kuo Kuang Rd., Taichung, 402, Taiwan

    Chuan-Mu Chen

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  1. Cheng-Wei Lai
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Corresponding author

Correspondence to Chuan-Mu Chen.

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Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

The animals used in this study were approved by the Institutional Animal Care and Use Committee (IACUC No. 103–97) of National Chung Hsing University, Taiwan.

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Lai, CW., Chen, HL., Yen, CC. et al. Using Dual Fluorescence Reporting Genes to Establish an In Vivo Imaging Model of Orthotopic Lung Adenocarcinoma in Mice. Mol Imaging Biol 18, 849–859 (2016). https://doi.org/10.1007/s11307-016-0967-4

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  • DOI: https://doi.org/10.1007/s11307-016-0967-4

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