Article Figures & Data
Figures
- Figure 1.
Samples of IR700-conjugated panitumumab (Pan-IR700) at concentrations of 2.5, 5, 20, and 40 μg/mL were prepared by diluting with PBS. A, fluorescence intensity image of Pan-IR700 solution. Fluorescence intensities were decreased according to decrease of concentration of Pan-IR700. B, fluorescence lifetime (FLT) image of Pan-IR700 solution. The FLT at various concentrations of Pan-IR700 solutions was almost the same value, 3.56 ± 0.081 nanoseconds (ns); 3.62 (2.5 μg/mL), 3.58 (5 μg/mL), 3.44 (20 μg/mL), 3.60 ns (40 μg/mL). C, LED light-irradiation for A431 cell pellets changes FLTs. A431 cell line incubated with Pan-IR700 for 24 hours were treated with PIT at doses of 0, 8, 15, and 30 J/cm2. FLT shortened to 3.09, 2.94, and 2.85 ns, compared with 3.28 ns before light exposure. These represented shortenings of 9.1%, 10.1%, and 13.1%, respectively. D, FLT of A431 pellets depends on the incubation time with Pan-IR700. FLT values escalate with incubation time with Pan-IR700. FLT changes from 2.98 ns (1 hour) to 3.42 ns (24 hours).
- Figure 2.
Serial fluorescence (bottom) and differential interference contrast microscopic images (top) of A431 cells, which were pre-incubated with Pan-IR700 (10 μg/mL) at 37°C for 24 hours, 5, 15, 60, and 90 seconds after start of exposing NIR light. (Pan-IR700 gradually internalized into cytoplasm in A431 cells after bound to cell membrane up to 24 hours after starting incubation.) Morphologic changes in differential interference contrast become severer with exposure to increased dose of NIR light. Scale bars, 50 μm.
- Figure 3.
Comparison with FLTs of irradiated tumors (dark gray bar) and nonirradiated tumors (bright gray bar). A, FLT images before and after PIT at the dose of 10, 30, and 50 J/cm2 in the same mouse that was inoculated with A431 cells on both sides of the mouse dorsum. Right-sided tumor was treated by PIT, whereas the left was covered. FLTs of A431 tumors treated with PIT with 50 J/cm2 (B), 30 J/cm2 (C), and 10 J/cm2 (D) were plotted. PIT with the NIR light dose of 30 and 50 J/cm2 showed significant (P < 0.05) shortenings in FLT immediately, compared with nonirradiated tumors. However, FLT did not significantly shorten at a low dose of 10 J/cm2. Transient prolongations of FLTs were observed at 6 hours after PIT likely due to uptake by reactive macrophages. Mann–Whitney's U test was used for the statistical analysis.
- Figure 4.
A, FLT in PIT treated tumors with 50 and 30 J/cm2 shortened significantly (P < 0.01) compared with no treatment control (0 J/cm2, control). FLTs were immediately shortened to 69.1 ± 10.9% and 61.5 ± 5.1% by PIT with 50 and 30 J/cm2, respectively. A431 tumors irradiated with only 10 J/cm2 showed no significant shortens of FLT that were seen immediately. FLT shortened by only 7.7% at 48 hours after PIT compared with no treatment control. B, FLT of nonirradiated tumors in PIT treated mice shortened slightly more than that in the untreated mice over time, but these changes were not significant. They may be caused by a small amount of light diffusing through tissue from the irradiated side, even though the surface of the tumor was covered. Student t test was used for the statistical analysis. C, histologic specimens of A431 tumors, which were treated with PIT at 0, 10, 30, and 50 J/cm2, are shown. All specimens were stained with hematoxylin and eosin. Microscopic evaluation of treated tumors revealed various degrees of necrosis and microhemorrhage with clusters of healthy or damaged tumor cells after PIT. Necrotic damage was diffuse and intense and fewer surviving tumor cells were seen when 30 and 50 J/cm2 of NIR light was administered. In contrast, when only 10 J/cm2 of NIR light was administered, necrotic cell damage was found in only limited areas within the tumor while substantial amounts of healthy cancer foci remained. Scale, 50 μm.
Additional Files
Supplementary Data
Files in this Data Supplement:
- Supplementary Methods - PDF file - 85K
- Supplementary Figure 1 - PDF file - 26K, Lapatinib induces a homologous recombination repair defect in HER2+ breast cancer cell
- Supplementary Figure 2 - PDF file - 25K, Homologous recombination repair capacity in MCF7 cells
- Supplementary Figure 3 - PDF file - 62K, MCF7 HER2 cells but not its isogenic MCF7 NEO controls are susceptible to the PARP inhibitors ABT-888 and AZD-2281
- Supplementary Figure 4 - PDF file - 45K, ABT-888 reduces levels of the NF�B regulated protein cMyc
- Supplementary Figure 5 - PDF file - 93K, Verification of the expected effects of p65 or I�B� overexpression on NFkappaB transcriptional activity in breast cancer cells
- Supplementary Table 1 - PDF file - 40K, Gene cluster, ER, PR and p53 status of the various HER2+ breast cancer cell lines ((3) and COSMIC database)
- Supplementary Legends for Table 1 and Figures 1-5 - PDF file - 95K
Volume 72, Issue 18
