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In vivo imaging of membrane type-1 matrix metalloproteinase with a novel activatable near-infrared fluorescence probe.

Shimizu Y, Temma T, Hara I, Makino A, Kondo N, Ozeki E, Ono M, Saji H - Cancer Sci. (2014)

Bottom Line: In aqueous buffer, MT1-hIC7L fluorescence was suppressed to background levels and increased approximately 35.5-fold in the presence of detergent.In MT1-MMP negative MCF-7 breast adenocarcinoma cells, both MT1-hIC7L and hIC7L showed no obvious fluorescence.In addition, the fluorescence intensity of C6 cells treated with MT1-hIC7L was suppressed by pre-treatment with an MT1-MMP endocytosis inhibitor (P < 0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Patho-Functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, Japan; Central Institute of Isotope Science, Hokkaido University, Sapporo, Japan.

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Cell uptake of MT1-hIC7L. (a,b) C6 (a) or MCF-7 cells (b) were treated with MT1-hIC7L or hIC7L and the fluorescence intensities were acquired for 6 h. Data are expressed as the FI ratio (mean ± SD) for 4 samples. Comparison between the MT1-hIC7L- and hIC7L-treated groups was performed with two-way factorial anova followed by Tukey–Kramer test (*P < 0.01 vs hIC7L). (c) The mean fluorescence intensity of C6 cells treated with (dotted line) or without Col I (solid line) for 6 h at 0.5, 1, 3 and 6 h after incubation with MT1-hIC7L. Data are expressed as%Intensity/mg protein (mean ± SD) for 4 samples. Comparisons between the Col I-treated (Col I [+]) and Col I-untreated groups (Col I [−]) were performed with two-way factorial anova followed by the Tukey–Kramer test (*P < 0.05 vs Col I [+]).
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fig03: Cell uptake of MT1-hIC7L. (a,b) C6 (a) or MCF-7 cells (b) were treated with MT1-hIC7L or hIC7L and the fluorescence intensities were acquired for 6 h. Data are expressed as the FI ratio (mean ± SD) for 4 samples. Comparison between the MT1-hIC7L- and hIC7L-treated groups was performed with two-way factorial anova followed by Tukey–Kramer test (*P < 0.01 vs hIC7L). (c) The mean fluorescence intensity of C6 cells treated with (dotted line) or without Col I (solid line) for 6 h at 0.5, 1, 3 and 6 h after incubation with MT1-hIC7L. Data are expressed as%Intensity/mg protein (mean ± SD) for 4 samples. Comparisons between the Col I-treated (Col I [+]) and Col I-untreated groups (Col I [−]) were performed with two-way factorial anova followed by the Tukey–Kramer test (*P < 0.05 vs Col I [+]).

Mentions: MT1-hIC7L fluorescence in C6 cells, which have high MT1-MMP expression levels, increased gradually and was significantly higher than that of hIC7L 3 h or more after addition (Fig. 3a). In contrast, probe uptake by MCF-7 cells (low MT1-MMP expression levels) was consistently low for both probes (Fig. 3b). When cells were pretreated with the endocytosis inhibitor Col I, the fluorescence intensity of cells incubated with MT1-hIC7L was significantly depressed compared to that of the non-pretreated group (Fig. 3c).


In vivo imaging of membrane type-1 matrix metalloproteinase with a novel activatable near-infrared fluorescence probe.

Shimizu Y, Temma T, Hara I, Makino A, Kondo N, Ozeki E, Ono M, Saji H - Cancer Sci. (2014)

Cell uptake of MT1-hIC7L. (a,b) C6 (a) or MCF-7 cells (b) were treated with MT1-hIC7L or hIC7L and the fluorescence intensities were acquired for 6 h. Data are expressed as the FI ratio (mean ± SD) for 4 samples. Comparison between the MT1-hIC7L- and hIC7L-treated groups was performed with two-way factorial anova followed by Tukey–Kramer test (*P < 0.01 vs hIC7L). (c) The mean fluorescence intensity of C6 cells treated with (dotted line) or without Col I (solid line) for 6 h at 0.5, 1, 3 and 6 h after incubation with MT1-hIC7L. Data are expressed as%Intensity/mg protein (mean ± SD) for 4 samples. Comparisons between the Col I-treated (Col I [+]) and Col I-untreated groups (Col I [−]) were performed with two-way factorial anova followed by the Tukey–Kramer test (*P < 0.05 vs Col I [+]).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4317866&req=5

fig03: Cell uptake of MT1-hIC7L. (a,b) C6 (a) or MCF-7 cells (b) were treated with MT1-hIC7L or hIC7L and the fluorescence intensities were acquired for 6 h. Data are expressed as the FI ratio (mean ± SD) for 4 samples. Comparison between the MT1-hIC7L- and hIC7L-treated groups was performed with two-way factorial anova followed by Tukey–Kramer test (*P < 0.01 vs hIC7L). (c) The mean fluorescence intensity of C6 cells treated with (dotted line) or without Col I (solid line) for 6 h at 0.5, 1, 3 and 6 h after incubation with MT1-hIC7L. Data are expressed as%Intensity/mg protein (mean ± SD) for 4 samples. Comparisons between the Col I-treated (Col I [+]) and Col I-untreated groups (Col I [−]) were performed with two-way factorial anova followed by the Tukey–Kramer test (*P < 0.05 vs Col I [+]).
Mentions: MT1-hIC7L fluorescence in C6 cells, which have high MT1-MMP expression levels, increased gradually and was significantly higher than that of hIC7L 3 h or more after addition (Fig. 3a). In contrast, probe uptake by MCF-7 cells (low MT1-MMP expression levels) was consistently low for both probes (Fig. 3b). When cells were pretreated with the endocytosis inhibitor Col I, the fluorescence intensity of cells incubated with MT1-hIC7L was significantly depressed compared to that of the non-pretreated group (Fig. 3c).

Bottom Line: In aqueous buffer, MT1-hIC7L fluorescence was suppressed to background levels and increased approximately 35.5-fold in the presence of detergent.In MT1-MMP negative MCF-7 breast adenocarcinoma cells, both MT1-hIC7L and hIC7L showed no obvious fluorescence.In addition, the fluorescence intensity of C6 cells treated with MT1-hIC7L was suppressed by pre-treatment with an MT1-MMP endocytosis inhibitor (P < 0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Patho-Functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, Japan; Central Institute of Isotope Science, Hokkaido University, Sapporo, Japan.

Show MeSH
Related in: MedlinePlus