Application of <i>in vivo</i> imaging system to establish and evaluate an experimental mouse model of lung cancer_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

CHEN Zhengju ¹ , GU Qiumei ¹ , XU Chunyan ¹ , LIU Zhiqiang ² , SHEN Meiling ¹ ,  SUN Huaiqiang ¹

1. Research Core Facilities, West China Hospital of Sichuan University, Chengdu, Sichuan 610041, P. R. China;
2. Precision Medicine Key Laboratory of Sichuan Province, West China Hospital of Sichuan University, Chengdu, Sichuan 610041, P. R. China;

Corresponding?author：

SUN Huaiqiang, Email: sunhuaiqiang@scu.edu.cn

Keywords：

Orthotopic lung cancer model; In vivo imaging system; A549 cells

DOI：

10.7507/1671-6205.202105060

Video：

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Objective To monitor the importance of establishing lung cancer models for immunological treatment through in vivo imaging system (IVIS). Methods In this study, a new optical bioluminescence IVIS was used to confirm the tumour formation and luminescence in male BALB/c nude mice by injecting A549-luc cells. First, A549-luc cells which expressed luciferase stably were transferred into nude mice by tail vein injection in order to establish a stable and reliable model of lung cancer. Then, D-fluorescein potassium salt was intraperitoneally injected every other week. The tumor formation and growth were dynamically observed on day 7^th, 14^th and 21^st by IVIS Spectrum and pathological exam with hematoxylin-eosin staining. Results Animal model of lung cancer was successfully established, and the development of lung cancer was effectively monitored by IVIS real-time fluorescence value which was consistent with pathological exam, and tumor volume was correlated with fluorescence intensity (r=0.7996, P<0.01). Conclusions IVIS has multiple benefits, including high sensitivity and specificity, simple operation, and no radiation. IVIS Spectrum can measure the fluorescence of tumor formed by injection of A549-luc cells in nude mice metastasis of lung cancer in a non-invasive, real-time and dynamic mode, which is worthy of promotion for using in clinical research.

Citation： CHEN Zhengju, GU Qiumei, XU Chunyan, LIU Zhiqiang, SHEN Meiling, SUN Huaiqiang. Application of in vivo imaging system to establish and evaluate an experimental mouse model of lung cancer. Chinese Journal of Respiratory and Critical Care Medicine, 2021, 20(11): 802-806. doi: 10.7507/1671-6205.202105060 Copy

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2.	Duma N, Santana-Davila R, Molina JR. Non-small cell lung cancer: epidemiology, screening, diagnosis, and treatment. Mayo Clin Proc, 2019, 94(8): 1623-1640.
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7.	胡麗麗, 阮永華. 移植性肺癌動物模型的建立和研究進展. 臨床與實驗病理學雜志, 2012, 28(8): 906-908.
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9.	Sierakowiak A, Henriques-Normark B, Iovino F. IVIS spectrum CT to image the progression of pneumococcal infections in vivo. Methods Mol Biol, 2019, 1968: 195-202.
10.	Tan X, Luo QL, Zhou SQ, et al. Erchen Plus Huiyanzhuyu decoction inhibits the growth of laryngeal carcinoma in a mouse model of phlegm-coagulation-blood-stasis syndrome via the STAT3/cyclin D1 pathway. Evid Based Complement Alternat Med, 2020, 2020: 2803496.
11.	Lofgren J, Miller AL, Lee CCS, et al. Analgesics promote welfare and sustain tumour growth in orthotopic 4 T1 and B16 mouse cancer models. Lab Anim, 2018, 52(4): 351-364.
12.	United Kingdom Co-ordinating Committee on Cancer Research (UKCCCR) Guidelines for the Welfare of Animals in Experimental Neoplasia (Second Edition). Br J Cancer, 1998, 77(1): 1-10.
13.	Huan Z, Zheng CL, Wang YZ, et al. miR-1323 promotes cell migration in lung adenocarcinoma by targeting Cbl-b and is an early prognostic biomarker. Front Oncol, 2020, 10: 181.
14.	Morita N, Haga S, Ohmiya Y, Ozaki M. Long-term ex vivo and in vivo monitoring of tumor progression by using dual luciferases. Anal Biochem, 2016, 497: 24-26.
15.	Yang WW, Lu Y, Xu YC, et al. Estrogen represses hepatocellular carcinoma (HCC) growth via inhibiting alternative activation of tumor-associated macrophages (TAMs). J Biol Chem, 2012, 287(48): 40140-40149.
16.	Zhang Q, Wang L, Qian Q, et al. Target area extraction algorithm for the in vivo fluorescence imaging of small animals. ACS Omega, 2020, 5(32): 20100-20106.
17.	Shrestha N, Lateef Z, Martey O, et al. Does the mouse tail vein injection method provide a good model of lung cancer?. F1000 Res, 2019, 8: 190.
18.	Keppens C, Dequeker EM, Pauwels P, et al. PD-L1 immunohistochemistry in non-small-cell lung cancer: unraveling differences in staining concordance and interpretation. Virchows Arch, 2021, 478(5): 827-839.
19.	劉啟才, 鄭國兵, 李曉艷, 等. 熒光素酶標記的肺癌細胞系的建立及動物模型驗證. 中國現代醫學雜志, 2011, 21(35): 4396-4399, 4404.
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21.	郭東勇, 李國友, 徐誠望, 等. 小動物活體成像法評價硝呋替莫對人源性神經母細胞瘤細胞SH-SY5Y腎轉移瘤模型裸鼠的治療作用. 國際藥學研究雜志, 2018, 45(11): 858-864.
22.	安國, 許輝, 張文龍, 等. 小動物活體成像系統Spectrum-CT在人肺癌裸鼠原位移植瘤模型建立中的應用. 腫瘤研究與臨床, 2020, 32(4): 250-255.
23.	李百慧. PI3 K抑制劑促進腫瘤血管正常化改善阿霉素治療乳腺癌的研究. 吉林大學, 2019.
24.	王偉, 陳超, 劉延明, 等. 應用小動物活體成像追蹤觀察EGFP標記的間充質干細胞. 中國細胞生物學學報, 2016, 38(2): 185-192.
25.	羅華灶, 李雪, 依含, 等. 熒光素酶標記小鼠結腸癌細胞建立活體成像移植瘤模型. 中國免疫學雜志, 2019, 35(17): 2106-2111.
26.	王蕾, 王曉英, 王玥, 等. 一種高效慢病毒轉染小鼠肺方法的建立. 軍事醫學, 2019, 43(8): 616-619.
27.	劉凡, 趙玉, 潘新宇, 等. 活體成像技術在NK/T細胞淋巴瘤早期檢測中的應用價值研究. 中國全科醫學, 2019, 22(30): 3689-3693,3700.

1. Wang YW, Zou SY, Zhao ZY, et al. New insights into small-cell lung cancer development and therapy. Cell Biol Int, 2020, 44(8): 1564-1576.
2. Duma N, Santana-Davila R, Molina JR. Non-small cell lung cancer: epidemiology, screening, diagnosis, and treatment. Mayo Clin Proc, 2019, 94(8): 1623-1640.
3. Balata H, Fong KM, Hendriks LE, et al. Prevention and early detection for NSCLC: advances in thoracic oncology 2018. J Thorac Oncol, 2019, 14(9): 1513-1527.
4. 周雪瑞, 黃選東. 免疫缺陷動物裸鼠. 生物學教學, 2007, 32(3): 2-4.
5. Malyla V, Paudel KR, Shukla SD, et al. Recent advances in experimental animal models of lung cancer. Future Med Chem, 2020, 12: 567-581.
6. Wang Y, Rouggly L, You M, et al. Animal models of lung cancer characterization and use for chemoprevention research. Prog Mol Biol Transl Sci, 2012, 105: 211-226.
7. 胡麗麗, 阮永華. 移植性肺癌動物模型的建立和研究進展. 臨床與實驗病理學雜志, 2012, 28(8): 906-908.
8. Lauber DT, Fül?p A, Kovács T, et al. State of the art in vivo imaging techniques for laboratory animals. Lab Anim, 2017, 51(5): 465-478.
9. Sierakowiak A, Henriques-Normark B, Iovino F. IVIS spectrum CT to image the progression of pneumococcal infections in vivo. Methods Mol Biol, 2019, 1968: 195-202.
10. Tan X, Luo QL, Zhou SQ, et al. Erchen Plus Huiyanzhuyu decoction inhibits the growth of laryngeal carcinoma in a mouse model of phlegm-coagulation-blood-stasis syndrome via the STAT3/cyclin D1 pathway. Evid Based Complement Alternat Med, 2020, 2020: 2803496.
11. Lofgren J, Miller AL, Lee CCS, et al. Analgesics promote welfare and sustain tumour growth in orthotopic 4 T1 and B16 mouse cancer models. Lab Anim, 2018, 52(4): 351-364.
12. United Kingdom Co-ordinating Committee on Cancer Research (UKCCCR) Guidelines for the Welfare of Animals in Experimental Neoplasia (Second Edition). Br J Cancer, 1998, 77(1): 1-10.
13. Huan Z, Zheng CL, Wang YZ, et al. miR-1323 promotes cell migration in lung adenocarcinoma by targeting Cbl-b and is an early prognostic biomarker. Front Oncol, 2020, 10: 181.
14. Morita N, Haga S, Ohmiya Y, Ozaki M. Long-term ex vivo and in vivo monitoring of tumor progression by using dual luciferases. Anal Biochem, 2016, 497: 24-26.
15. Yang WW, Lu Y, Xu YC, et al. Estrogen represses hepatocellular carcinoma (HCC) growth via inhibiting alternative activation of tumor-associated macrophages (TAMs). J Biol Chem, 2012, 287(48): 40140-40149.
16. Zhang Q, Wang L, Qian Q, et al. Target area extraction algorithm for the in vivo fluorescence imaging of small animals. ACS Omega, 2020, 5(32): 20100-20106.
17. Shrestha N, Lateef Z, Martey O, et al. Does the mouse tail vein injection method provide a good model of lung cancer?. F1000 Res, 2019, 8: 190.
18. Keppens C, Dequeker EM, Pauwels P, et al. PD-L1 immunohistochemistry in non-small-cell lung cancer: unraveling differences in staining concordance and interpretation. Virchows Arch, 2021, 478(5): 827-839.
19. 劉啟才, 鄭國兵, 李曉艷, 等. 熒光素酶標記的肺癌細胞系的建立及動物模型驗證. 中國現代醫學雜志, 2011, 21(35): 4396-4399, 4404.
20. 樊志超. 應用活體流式細胞儀和活體成像技術對肝癌進展與轉移的監測及療效評估研究. 復旦大學, 2013.
21. 郭東勇, 李國友, 徐誠望, 等. 小動物活體成像法評價硝呋替莫對人源性神經母細胞瘤細胞SH-SY5Y腎轉移瘤模型裸鼠的治療作用. 國際藥學研究雜志, 2018, 45(11): 858-864.
22. 安國, 許輝, 張文龍, 等. 小動物活體成像系統Spectrum-CT在人肺癌裸鼠原位移植瘤模型建立中的應用. 腫瘤研究與臨床, 2020, 32(4): 250-255.
23. 李百慧. PI3 K抑制劑促進腫瘤血管正常化改善阿霉素治療乳腺癌的研究. 吉林大學, 2019.
24. 王偉, 陳超, 劉延明, 等. 應用小動物活體成像追蹤觀察EGFP標記的間充質干細胞. 中國細胞生物學學報, 2016, 38(2): 185-192.
25. 羅華灶, 李雪, 依含, 等. 熒光素酶標記小鼠結腸癌細胞建立活體成像移植瘤模型. 中國免疫學雜志, 2019, 35(17): 2106-2111.
26. 王蕾, 王曉英, 王玥, 等. 一種高效慢病毒轉染小鼠肺方法的建立. 軍事醫學, 2019, 43(8): 616-619.
27. 劉凡, 趙玉, 潘新宇, 等. 活體成像技術在NK/T細胞淋巴瘤早期檢測中的應用價值研究. 中國全科醫學, 2019, 22(30): 3689-3693,3700.

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Chinese Journal of Respiratory and Critical Care Medicine

Application of in vivo imaging system to establish and evaluate an experimental mouse model of lung cancer

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