Journal of the Korean Society of Radiology (대한영상의학회지)

The Korean Society of Radiology (대한영상의학회)
권호 표지

A Bone Metastasis Nude Mouse Model Created by Ultrasound Guided Intracardiac Injection of Breast Cancer Cells: the Micro-CT, MRI and Bioluminescence Imaging Analysis

누드 마우스에서 초음파 유도하의 심장 내 유방암세포 주입을 통한 골전이암 모델 생성과 미세전산화단층촬영, 자기공명영상, 및 생물발광영상 분석

  • Park, Young-Jin (Department of Radiology, Research Institute of Radiological Science, College of Medicine, Yonsei University) ;
  • Song, Eun-Hye (Department of Radiology, Research Institute of Radiological Science, College of Medicine, Yonsei University) ;
  • Kim, Seol-Hwa (Department of Radiology, Research Institute of Radiological Science, College of Medicine, Yonsei University) ;
  • Choi, Sang-Hyun (Korean Minjok Leadership Academy) ;
  • Song, Ho-Taek (Department of Radiology, Research Institute of Radiological Science, College of Medicine, Yonsei University) ;
  • Suh, Jin-Suck (Department of Radiology, Research Institute of Radiological Science, College of Medicine, Yonsei University)
  • 박영진 (연세대학교 의과대학 영상의학교실 방사선의과학연구소) ;
  • 송은혜 (연세대학교 의과대학 영상의학교실 방사선의과학연구소) ;
  • 김설화 (연세대학교 의과대학 영상의학교실 방사선의과학연구소) ;
  • 최상현 (민족사관고등학교) ;
  • 송호택 (연세대학교 의과대학 영상의학교실 방사선의과학연구소) ;
  • 서진석 (연세대학교 의과대학 영상의학교실 방사선의과학연구소)
  • Published : 2011.01.01
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Abstract

Purpose: The purpose of this study was to develop a nude mouse model of bone metastasis by performing intracardiac injection of breast cancer cells under ultrasonography guidance and we wanted to evaluate the development and the distribution of metastasis in vivo using micro-CT, MRI and bioluminescence imaging. Materials and Methods: Animal experiments were performed in 6-week-old female nude mice. The animals underwent left ventricular injection of $2{\times}10^{5}$ MDA-MB-231Bo-Luc cells. After injection of the tumor cells, serial bioluminescence imaging was performed for 7 weeks. The findings of micro-CT, MRI and the histology were correlated with the 'hot' lesions seen on the bioluminescence imaging. Results: Metastasis was found in 62.3% of the animals. Two weeks after intracardiac injection, metastasis to the brain, spine and femur was detected with bioluminescence imaging with an increasing intensity by week 7. Micro-CT scan confirmed multiple osteolytic lesions at the femur, spine and skull. MRI and the histology were able to show metastasis in the brain and extraskeletal metastasis around the femur. Conclusion: The intracardiac injection of cancer cells under ultrasonography guidance is a safe and highly reproducible method to produce bone metastasis in nude mice. This bone metastasis nude mouse model will be useful to study the mechanism of bone metastasis and to validate new therapeutics.

목적: 누드 마우스에서 고해상도 초음파 유도 하에 암세포를 심장 내 주입하는 방법을 통한 유방암의 골전이 모델을 생성하고자 하였으며 생체 내에서의 골전이암의 발생과 분포 및 증식을 미세전산화단층촬영, 자기공명영상, 그리고 생물발광영상을 이용하여 평가하고자 하였다. 대상과 방법: 6주령된 누드 마우스를 호흡 마취한 후 $2{\times}10^{5}$개의 MDA-MB-231Bo-Luc 세포를 17MHz 고해상도 초음파 유도 하에 좌심실에 주입하였다. 주입 후 7주 동안 생물발광영상을 하였으며, 7주째에 미세전산화단층촬영과 자기공명영상으로 골용해 병변과 연부조직 전이암을 분석하였다. 영상에서 전이가 확인된 부위의 조직 슬라이드 절편을 얻고 헤마톡실린 에오신 (hematoxylin-eosin) 염색과 cytokeratin 면역염색을 시행하여 현미경으로 관찰하였다. 결과: 전이암 생성율은 62.3%(5/8)이었다. 두부, 대퇴골부위와 척추에서 강한 생물발광 신호가 측정되었으며 2주부터 7주까지 세기가 증가하였다. 미세전산화단층촬영에서는 생물발광영상과 일치하는 두개골과 대퇴골 및 척추의 골용해 병변이 확인되었으며 자기공명영상과 조직 현미경 검사에서 대퇴골 주위로 자라난 종양과 소뇌에 발생한 전이암이 확인되었다. 결론: 초음파 유도하의 심장 내 종양세포 주입은 안전하고 높은 재현성으로 유방암의 골전이 모델을 형성할 방법으로서 이 동물 모델은 골전이암의 기전 연구와 치료제 평가에 유용하리라 생각한다.

Keywords

References

  1. Yoneda T. Arterial microvascularization and breast cancer colonization in bone. Histol Histopathol 1997;12:1145-1149
  2. Yoneda T, Sasaki A, Mundy GR. Osteolytic bone metastasis in breast cancer. Breast Cancer Res Treat 1994;32:73-84 https://doi.org/10.1007/BF00666208
  3. Arguello F, Baggs RB, Frantz CN. A murine model of experimental metastasis to bone and bone marrow. Cancer Res 1988;48:6876-6881
  4. Guise TA, Yin JJ, Taylor SD, Kumagai Y, Dallas M, Boyce BF, et al. Evidence for a causal role of parathyroid hormone-related protein in the pathogenesis of human breast cancer-mediated osteolysis. J Clin Invest 1996;98:1544-1549 https://doi.org/10.1172/JCI118947
  5. Corey E, Quinn JE, Bladou F, Brown LG, Roudier MP, Brown JM, et al. Establishment and characterization of osseous prostate cancer models: intra-tibial injection of human prostate cancer cells. Prostate 2002;52:20-33 https://doi.org/10.1002/pros.10091
  6. Wang CY, Chang YW. A model for osseous metastasis of human breast cancer established by intrafemur injection of the mda-mb- 435 cells in nude mice. Anticancer Res 1997;17:2471-2474
  7. Kjonniksen I, Winderen M, Bruland O, Fodstad O. Validity and usefulness of human tumor models established by intratibial cell inoculation in nude rats. Cancer Res 1994;54:1715-1719
  8. Peterschmitt J, Bauerle T, Berger MR. Effect of zoledronic acid and an antibody against bone sialoprotein II on MDA-MB-231(GFP) breast cancer cells in vitro and on osteolytic lesions induced in vivo by this cell line in nude rats. Clin Exp Metastasis 2007;24:449-459 https://doi.org/10.1007/s10585-007-9082-x
  9. Bauerle T, Adwan H, Kiessling F, Hilbig H, Armbruster FP, Berger MR. Characterization of a rat model with site-specific bone metastasis induced by MDA-MB-231 breast cancer cells and its application to the effects of an antibody against bone sialoprotein. Int J Cancer 2005;115:177-186 https://doi.org/10.1002/ijc.20840
  10. Hsu WK, Virk MS, Feeley BT, Stout DB, Chatziioannou AF, Lieberman JR. Characterization of osteolytic, osteoblastic, and mixed lesions in a prostate cancer mouse model using 18F-FDG and 18F-Fluoride PET/CT. J Nucl Med 2008;49:414-421 https://doi.org/10.2967/jnumed.107.045666
  11. Jenkins DE, Hornig YS, Oei Y, Dusich J, Purchio T. Bioluminescent human breast cancer cell lines that permit rapid and sensitive in vivo detection of mammary tumors and multiple metastases in immune deficient mice. Breast Cancer Res 2005;7:R444-R454 https://doi.org/10.1186/bcr1026
  12. Minn AJ, Kang Y, Serganova I, Gupta GP, Giri DD, Doubrovin M, et al. Distinct organ-specific metastatic potential of individual breast cancer cells and primary tumors. J Clin Invest 2005;115:44-55 https://doi.org/10.1172/JCI22320
  13. Wetterwald A, van der Pluijm G, Que I, Sijmons B, Buijs J, Karperien M, et al. Optical imaging of cancer metastasis to bone marrow: a mouse model of minimal residual disease. Am J Pathol 2002;160:1143-1153 https://doi.org/10.1016/S0002-9440(10)64934-6
  14. Song H, Shahverdi K, Huso DL, Wang Y, Fox JJ, Hobbs RF, et al. An immunotolerant HER-2/neu transgenic mouse model of metastatic breast cancer. Clin Cancer Res 2008;14:6116-6124 https://doi.org/10.1158/1078-0432.CCR-07-4672
  15. Mouchess ML, Sohara Y, Nelson MD Jr, DeCLerck YA, Moats RA. Multimodal imaging analysis of tumor progression and bone resorption in a murine cancer model. J Comput Assist Tomogr 2006;30:525-534 https://doi.org/10.1097/00004728-200605000-00030
  16. Yoneda T, Williams PJ, Hiraga T, Niewolna M, Nishimura R. A bone-seeking clone exhibits different biological properties from the MDA-MB-231 parental human breast cancer cells and a brainseeking clone in vivo and in vitro. J Bone Miner Res 2001;16:1486-1495 https://doi.org/10.1359/jbmr.2001.16.8.1486
  17. Jonkers J, Derksen PW. Modeling metastatic breast cancer in mice. J Mammary Gland Biol 2007;12:191-203 https://doi.org/10.1007/s10911-007-9050-8
  18. Hortobagyi GN. Developments in chemotherapy of breast cancer. Cancer 2000;88(12 Suppl):3073-3079 https://doi.org/10.1002/1097-0142(20000615)88:12+<3073::AID-CNCR26>3.0.CO;2-R
  19. Liotta LA, Kohn EC. The microenvironment of the tumour-host interface. Nature 2001;411:375-379 https://doi.org/10.1038/35077241
  20. Kim S. Animal models of cancer in the head and neck region. Clin Exp Otorhinolar 2009;2:55-60 https://doi.org/10.3342/ceo.2009.2.2.55
  21. Rosol TJ, Tannehill-Gregg SH, Corn S, Schneider A, McCauley LK. Animal models of bone metastasis. Cancer Treat Res 2004;118:47-81
  22. Sano D, Myers JN. Xenograft models of head and neck cancers. Head Neck Oncol 2009;1:32 https://doi.org/10.1186/1758-3284-1-32
  23. Rozel S, Galban CJ, Nicolay K, Lee KC, Sud S, Neeley C, et al. Synergy between anti-CCL2 and docetaxel as determined by DWMRI in a metastatic bone cancer model. J Cell Biochem 2009;107:58-64 https://doi.org/10.1002/jcb.22056
  24. Song HT, Jordan EK, Lewis BK, Liu W, Ganjei J, Klaunberg B, et al. Rat model of metastatic breast cancer monitored by MRI at 3 tesla and bioluminescence imaging with histological correlation. J Transl Med 2009;7:88 https://doi.org/10.1186/1479-5876-7-88