Establishment and evaluation of a new highly metastatic tumor cell line 5a-D-Luc-ZsGreen expressing both luciferase and green fluorescent protein Hitomi Sudo1

Atsushi B. Tsuji, Aya Sugyo, Hiroyuki Takuwa, Kazuto Masamoto, Yutaka Tomita, Norihiro Suzuki, Takeshi Imamura, Mitsuru Koiz Umi, Tsuneo Saga

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Breast cancer is the most common cancer in women. Although advances in diagnostic imaging for early detection, surgical techniques and chemotherapy have improved overall survival, the prognosis of patients with metastatic breast cancer remains poor. Understanding cancer cell dynamics in the metastatic process is important to develop new therapeutic strategies. Experimental animal models and imaging would be powerful tools for understanding of the molecular events of multistep process of metastasis. In the present study, to develop a new cancer cell line that is applicable to bioluminescence and fluorescence imaging, we transfected the expression vector of a green fluorescent protein ZsGreen1 into a metastatic cell line 5a-D-Luc, which is a subclone of the MDA-MB-231 breast cancer cell line expressing luciferase, and established a new tumor cell line 5a-D-Luc-ZsGreen expressing both luciferase and ZsGreen1. The 5a-D-Luc-ZsGreen cells proliferate more rapidly and have a more invasive phenotype compared with 5a-D-Luc cells following intracardiac injection. Metastasis sites were easily detected in the whole body by bioluminescence imaging and in excised tissues by ex vivo fluorescence imaging. The fluorescence of 5a-D-Luc-ZsGreen cells was not lost after formalin fixation and decalcification. It enabled us to easily evaluate tumor spread and localization at the cellular level in microscopic analysis. The strong fluorescence of 5a-DLuc- ZsGreen cells allowed for real-time imaging of circulating tumor cells in cerebral blood vessels of live animals immediately after intracardiac injection of cells using two-photon laser-scanning microscopy. These findings suggest that the 5a-D-Luc-ZsGreen cells would be a useful tool for research on mechanisms of metastatic process in animal models.

Original languageEnglish
Pages (from-to)525-532
Number of pages8
JournalInternational Journal of Oncology
Volume48
Issue number2
DOIs
Publication statusPublished - 2016 Feb 1

Fingerprint

Green Fluorescent Proteins
Tumor Cell Line
Luciferases
Optical Imaging
Breast Neoplasms
Cell Line
Neoplasms
Animal Models
Fluorescence
Neoplasm Metastasis
Circulating Neoplastic Cells
Whole Body Imaging
Injections
Diagnostic Imaging
Photons
Confocal Microscopy
Formaldehyde
Blood Vessels
Phenotype
Drug Therapy

Keywords

  • Animal model
  • Bioluminescence imaging
  • Bone metastasis
  • Brain metastasis
  • Fluorescence imaging
  • In vivo imaging
  • Real-time imaging
  • Two-photon excitation microscopy

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Establishment and evaluation of a new highly metastatic tumor cell line 5a-D-Luc-ZsGreen expressing both luciferase and green fluorescent protein Hitomi Sudo1. / Tsuji, Atsushi B.; Sugyo, Aya; Takuwa, Hiroyuki; Masamoto, Kazuto; Tomita, Yutaka; Suzuki, Norihiro; Imamura, Takeshi; Koiz Umi, Mitsuru; Saga, Tsuneo.

In: International Journal of Oncology, Vol. 48, No. 2, 01.02.2016, p. 525-532.

Research output: Contribution to journalArticle

Tsuji, Atsushi B. ; Sugyo, Aya ; Takuwa, Hiroyuki ; Masamoto, Kazuto ; Tomita, Yutaka ; Suzuki, Norihiro ; Imamura, Takeshi ; Koiz Umi, Mitsuru ; Saga, Tsuneo. / Establishment and evaluation of a new highly metastatic tumor cell line 5a-D-Luc-ZsGreen expressing both luciferase and green fluorescent protein Hitomi Sudo1. In: International Journal of Oncology. 2016 ; Vol. 48, No. 2. pp. 525-532.
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abstract = "Breast cancer is the most common cancer in women. Although advances in diagnostic imaging for early detection, surgical techniques and chemotherapy have improved overall survival, the prognosis of patients with metastatic breast cancer remains poor. Understanding cancer cell dynamics in the metastatic process is important to develop new therapeutic strategies. Experimental animal models and imaging would be powerful tools for understanding of the molecular events of multistep process of metastasis. In the present study, to develop a new cancer cell line that is applicable to bioluminescence and fluorescence imaging, we transfected the expression vector of a green fluorescent protein ZsGreen1 into a metastatic cell line 5a-D-Luc, which is a subclone of the MDA-MB-231 breast cancer cell line expressing luciferase, and established a new tumor cell line 5a-D-Luc-ZsGreen expressing both luciferase and ZsGreen1. The 5a-D-Luc-ZsGreen cells proliferate more rapidly and have a more invasive phenotype compared with 5a-D-Luc cells following intracardiac injection. Metastasis sites were easily detected in the whole body by bioluminescence imaging and in excised tissues by ex vivo fluorescence imaging. The fluorescence of 5a-D-Luc-ZsGreen cells was not lost after formalin fixation and decalcification. It enabled us to easily evaluate tumor spread and localization at the cellular level in microscopic analysis. The strong fluorescence of 5a-DLuc- ZsGreen cells allowed for real-time imaging of circulating tumor cells in cerebral blood vessels of live animals immediately after intracardiac injection of cells using two-photon laser-scanning microscopy. These findings suggest that the 5a-D-Luc-ZsGreen cells would be a useful tool for research on mechanisms of metastatic process in animal models.",
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AU - Tsuji, Atsushi B.

AU - Sugyo, Aya

AU - Takuwa, Hiroyuki

AU - Masamoto, Kazuto

AU - Tomita, Yutaka

AU - Suzuki, Norihiro

AU - Imamura, Takeshi

AU - Koiz Umi, Mitsuru

AU - Saga, Tsuneo

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AB - Breast cancer is the most common cancer in women. Although advances in diagnostic imaging for early detection, surgical techniques and chemotherapy have improved overall survival, the prognosis of patients with metastatic breast cancer remains poor. Understanding cancer cell dynamics in the metastatic process is important to develop new therapeutic strategies. Experimental animal models and imaging would be powerful tools for understanding of the molecular events of multistep process of metastasis. In the present study, to develop a new cancer cell line that is applicable to bioluminescence and fluorescence imaging, we transfected the expression vector of a green fluorescent protein ZsGreen1 into a metastatic cell line 5a-D-Luc, which is a subclone of the MDA-MB-231 breast cancer cell line expressing luciferase, and established a new tumor cell line 5a-D-Luc-ZsGreen expressing both luciferase and ZsGreen1. The 5a-D-Luc-ZsGreen cells proliferate more rapidly and have a more invasive phenotype compared with 5a-D-Luc cells following intracardiac injection. Metastasis sites were easily detected in the whole body by bioluminescence imaging and in excised tissues by ex vivo fluorescence imaging. The fluorescence of 5a-D-Luc-ZsGreen cells was not lost after formalin fixation and decalcification. It enabled us to easily evaluate tumor spread and localization at the cellular level in microscopic analysis. The strong fluorescence of 5a-DLuc- ZsGreen cells allowed for real-time imaging of circulating tumor cells in cerebral blood vessels of live animals immediately after intracardiac injection of cells using two-photon laser-scanning microscopy. These findings suggest that the 5a-D-Luc-ZsGreen cells would be a useful tool for research on mechanisms of metastatic process in animal models.

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KW - Real-time imaging

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