Open AccessLive imaging of transforming growth factor‐β activated kinase 1 activation in Lewis lung carcinoma 3 LL cells implanted into syngeneic mice and treated with polyinosinic:polycytidylic acid
Author(s) -
Takaoka Saori,
Kamioka Yuji,
Takakura Kanako,
Baba Ai,
Shime Hiroaki,
Seya Tsukasa,
Matsuda Michiyuki
Publication year - 2016
Publication title -
cancer science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.035
H-Index - 141
eISSN - 1349-7006
pISSN - 1347-9032
DOI - 10.1111/cas.12923
Subject(s) - lewis lung carcinoma , tumor necrosis factor alpha , cancer research , chemistry , anisomycin , transforming growth factor , kinase , microbiology and biotechnology , medicine , biology , cancer , biochemistry , metastasis
Transforming growth factor‐β activated kinase 1 ( TAK 1) has been shown to play a crucial role in cell death, differentiation, and inflammation. Here, we live‐imaged robust TAK 1 activation in Lewis lung carcinoma 3 LL cells implanted into the s.c. tissue of syngeneic C57 BL /6 mice and treated with polyinosinic:polycytidylic acid (PolyI:C). First, we developed and characterized a Förster resonance energy transfer‐based biosensor for TAK 1 activity. The TAK 1 biosensor, named Eevee‐ TAK 1, responded to stress‐inducing reagents such as anisomycin, tumor necrosis factor‐α, and interleukin1‐β. The anisomycin‐induced increase in Förster resonance energy transfer was abolished by the TAK 1 inhibitor (5z)‐7‐oxozeaenol. Activity of TAK 1 in 3 LL cells was markedly increased by PolyI:C in the presence of macrophages. 3 LL cells expressing Eevee‐ TAK 1 were implanted into mice and observed through imaging window by two‐photon excitation microscopy. During the growth of tumor, the 3 LL cells at the periphery of the tumor showed higher TAK 1 activity than the 3 LL cells located at the center of the tumor, suggesting that cells at the periphery of the tumor mass were under stronger stress. Injection of PolyI:C, which is known to induce regression of the implanted tumors, induced marked and homogenous TAK 1 activation within the tumor tissues. The effect of PolyI:C faded within 4 days. These observations suggest that Eevee‐ TAK 1 is a versatile tool to monitor cellular stress in cancer tissues.