TY - JOUR
T1 - Intracellular pH imaging in cancer cells in vitro and tumors in vivo using the new genetically encoded sensor SypHer2
AU - Shirmanova, Marina V.
AU - Druzhkova, Irina N.
AU - Lukina, Maria M.
AU - Matlashov, Mikhail E.
AU - Belousov, Vsevolod V.
AU - Snopova, Ludmila B.
AU - Prodanetz, Natalia N.
AU - Dudenkova, Varvara V.
AU - Lukyanov, Sergey A.
AU - Zagaynova, Elena V.
N1 - Funding Information:
This work was supported by the Russian Science Foundation (project # 14-15-00646 ). The authors are grateful to Natalia Klementieva for her kind assistance with the Nikon Eclipse TI N-STORM microscope.
Publisher Copyright:
© 2015 Elsevier B.V.
PY - 2015/6/12
Y1 - 2015/6/12
N2 - Background Measuring intracellular pH (pHi) in tumors is essential for the monitoring of cancer progression and the response of cancer cells to various treatments. The purpose of the study was to develop a method for pHi mapping in living cancer cells in vitro and in tumors in vivo, using the novel genetically encoded indicator, SypHer2. Methods A HeLa Kyoto cell line stably expressing SypHer2 in the cytoplasm was used, to perform ratiometric (dual excitation) imaging of the probe in cell culture, in 3D tumor spheroids and in tumor xenografts in living mice. Results Using SypHer2, pHi was demonstrated to be 7.34 ± 0.11 in monolayer HeLa cells in vitro under standard cultivation conditions. An increasing pHi gradient from the center to the periphery of the spheroids was displayed. We obtained fluorescence ratio maps for HeLa tumors in vivo and ex vivo. Comparison of the map with the pathomorphology and with hypoxia staining of the tumors revealed a correspondence of the zones with higher pHi to the necrotic and hypoxic areas. Conclusions Our results demonstrate that pHi imaging with the genetically encoded pHi indicator, SypHer2, can be a valuable tool for evaluating tumor progression in xenograft models. General significance We have demonstrated, for the first time, the possibility of using the genetically encoded sensor SypHer2 for ratiometric pH imaging in cancer cells in vitro and in tumors in vivo. SypHer2 shows great promise as an instrument for pHi monitoring able to provide high accuracy and spatiotemporal resolution.
AB - Background Measuring intracellular pH (pHi) in tumors is essential for the monitoring of cancer progression and the response of cancer cells to various treatments. The purpose of the study was to develop a method for pHi mapping in living cancer cells in vitro and in tumors in vivo, using the novel genetically encoded indicator, SypHer2. Methods A HeLa Kyoto cell line stably expressing SypHer2 in the cytoplasm was used, to perform ratiometric (dual excitation) imaging of the probe in cell culture, in 3D tumor spheroids and in tumor xenografts in living mice. Results Using SypHer2, pHi was demonstrated to be 7.34 ± 0.11 in monolayer HeLa cells in vitro under standard cultivation conditions. An increasing pHi gradient from the center to the periphery of the spheroids was displayed. We obtained fluorescence ratio maps for HeLa tumors in vivo and ex vivo. Comparison of the map with the pathomorphology and with hypoxia staining of the tumors revealed a correspondence of the zones with higher pHi to the necrotic and hypoxic areas. Conclusions Our results demonstrate that pHi imaging with the genetically encoded pHi indicator, SypHer2, can be a valuable tool for evaluating tumor progression in xenograft models. General significance We have demonstrated, for the first time, the possibility of using the genetically encoded sensor SypHer2 for ratiometric pH imaging in cancer cells in vitro and in tumors in vivo. SypHer2 shows great promise as an instrument for pHi monitoring able to provide high accuracy and spatiotemporal resolution.
KW - Cancer cell
KW - Genetically encoded sensor
KW - Intracellular pH
KW - Ratiometric imaging
KW - SypHer2
KW - Tumor
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U2 - 10.1016/j.bbagen.2015.05.001
DO - 10.1016/j.bbagen.2015.05.001
M3 - Article
C2 - 25964069
AN - SCOPUS:84930959637
SN - 0304-4165
VL - 1850
SP - 1905
EP - 1911
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
IS - 9
ER -
