Journal: Chemical Science

Article Title: Light emission enhancement by supramolecular complexation of chemiluminescence probes designed for bioimaging

doi: 10.1039/c8sc05174g

Figure Lengend Snippet: Figure 7: A) Chemical structures of probe 2 and TMCD-FITC. B) Chemiluminescence imaging and (C) quantification of signal intensities in HEK293-Lac-Z cells incubated with the complex of probe 2 [10 μM] and TMCD-FITC [50 μM]. (D) (a’) Transmitted light image and (b’) chemiluminescence microscopy image of HEK293-LacZ cells. (c’) Transmitted light image and (d’) chemiluminescence microscopy image of HEK293-WT cells. Images were obtained following 20 min incubation with cell culture medium containing probe 2 [50 μM] and TMCD [50 μM]. Images were taken using the LV200 Olympus microscope using a 60× objective and 5 min exposure time.

Article Snippet: Images were taken using the LV200 Olympus microscope using a 60× objective and 5 min exposure time.

Techniques: Imaging, Incubation, Microscopy, Cell Culture

Journal: eBioMedicine

Article Title: Sialic acid blockade inhibits the metastatic spread of prostate cancer to bone

doi: 10.1016/j.ebiom.2024.105163

Figure Lengend Snippet: Upregulation of ST6GAL1 promotes the spread of prostate cancer to bone. ( a ) Luciferase tagged PC3 cells (control or ST6GAL1 overexpressing) were injected into BALB/c nude mice via intra cardiac injection. ( b ) Tumours were monitored over 6 weeks using in vivo bioluminescence imaging. Upregulation of ST6GAL1 significantly increased the number of skeletal tumours formed per mouse (n = 10, unpaired t test, p = 0.006). ( c ) Ex vivo micro-CT analysis was used to examine tumour induced bone destruction in left tibias and showed that overexpression of ST6GAL1 in PC3 cells significantly enhanced tumour incidence (representative images are shown) (n = 10, Chi-square test, p = 0.04). Scale bar is 1 mm. ( d , e ) Further bone histomorphometry demonstrated that in mice bearing ST6GAL1 overexpressing tumours there were significantly higher number of osteoclasts per mm medial bone surface (n = 4 unpaired t test, p = 0.0055) but lower number of osteoblasts per mm lateral bone surface in tibias without overt tumours, compared to mice bearing control tumours (n = 5 unpaired t test, p = 0.0452). ( f ) Sandwich ELISA analysis of CSF1 levels in conditioned media samples from PC3 prostate cancer cells with overexpression of ST6GAL1. CSF1 levels are significantly increased in conditioned media samples from PC3 cells with upregulation of ST6GAL1 (unpaired t test, p = 0.003). ( g ) Conditioned media samples from DU145 cells overexpressing ST6GAL1 promotes the differentiation of murine primary osteoclast pre-cursors into osteoclasts measured via TRAP staining (unpaired t test, p = 0.0005). Scale bar is 100 μm. ( h , i ) Primary monocytes from healthy controls were treated with 150 μl of concentrated conditioned media from PC3 cells (control or ST6GAL1 overexpressing) at days 0 and 3 in the presence of 5 μg/ml anti-MCSF or isotype control. Cells were harvested on day 6 and assessed for viability (h) and CD206 expression (i) using flow cytometry. (n = 5, two-way ANOVA, multiple comparisons, (h) cont. vs. OE isotype. p = 0.002. OE isotype vs. OE anti-MCSF p = 0.028. (i) cont. vs OE isotype. p < 0.0001. cont. vs OE isotype. p = 0.0002).

Article Snippet: Tumours were monitored over 15 days using in vivo bioluminescence imaging. ( g , h ) Pre-treatment of RM1 cells with P-SiaFNEtoc (to remove sialylated glycans) significantly reduced the number of skeletal tumours formed (Mann–Whitney test, p = 0.0454), the incidence of tumour in left tibias (Chi-square test, p = 0.0455), and significantly increased survival time in mice (Log-rank test, p = 0.012). ( i ) Micro-CT analysis demonstrated that P-SiaFNEtoc significantly alleviated bone destruction in the trabecular bone of tibias and increased trabecular bone volume (BV/TV, p = 0.0211) and trabecular number (Tb.

Techniques: Luciferase, Control, Injection, In Vivo, Imaging, Ex Vivo, Micro-CT, Over Expression, Sandwich ELISA, Staining, Expressing, Flow Cytometry

Journal: eBioMedicine

Article Title: Sialic acid blockade inhibits the metastatic spread of prostate cancer to bone

doi: 10.1016/j.ebiom.2024.105163

Figure Lengend Snippet: Sialic acid blockade can prevent/inhibit prostate cancer bone metastasis. ( a ) Inhibition of sialylation in TRAMPC2 cells using P-SiaFNEtoc detected using pan-specific Lectenz lectin flow cytometry. Cells were treated with a range of concentrations of P-SiaFNEtoc inhibitor from 2 μM to 512 μM for 72 h. The intensities were normalised to a DMSO control. ( b ) Detection of α2-6 linked sialylated N -glycans in TRAMPC2 cells using SNA lectin flow cytometry. TRAMPC2 cells treated with 64 μM P-SiaFNEtoc for 72 h had reduced levels of SNA binding indicating a reduction in α2-6 linked sialylation in these cells (unpaired t test, p = 0.0001). ( c ) Luciferase tagged TRAMPC2 cells (control or pre-treated with 64 μM P-SiaFNEtoc for 72 h) were injected into immunocompetent C57BL/6 mice via sub-cutaneous injection and tumours were monitored using in vivo bioluminescence imaging. Pre-treatment of TRAMPC2 cells with P-SiaFNEtoc (which removed sialylated glycans) significantly reduced tumour burden over 6 weeks (n = 10, Mann–Whitney test, p = 0.0233) thus suggesting that sialic acid blockade has the potential to inhibit the growth of prostate tumours. ( d ) Inhibition of sialylation in RM1 cells using P-SiaFNEtoc detected using pan-specific Lectenz lectin flow cytometry. Cells were treated with a range of concentrations of P-SiaFNEtoc inhibitor from 2 μM to 512 μM for 72 h. The intensities were normalised to a DMSO control. ( e ) Detection of α2-6 linked sialylated N -glycans in RM1 cells using SNA lectin flow cytometry. RM1 cells treated with 256 μM P-SiaFNEtoc for 72 h had reduced levels of SNA binding indicating a reduction in α2-6 linked sialylation in these cells (unpaired t test, p < 0.0001). ( f ) Luciferase tagged RM1 cells (control or pre-treated with 256 μM P-SiaFNEtoc for 72 h) were injected into immunocompetent C57BL/6 mice via intra cardiac injection. Tumours were monitored over 15 days using in vivo bioluminescence imaging. ( g , h ) Pre-treatment of RM1 cells with P-SiaFNEtoc (to remove sialylated glycans) significantly reduced the number of skeletal tumours formed (Mann–Whitney test, p = 0.0454), the incidence of tumour in left tibias (Chi-square test, p = 0.0455), and significantly increased survival time in mice (Log-rank test, p = 0.012). ( i ) Micro-CT analysis demonstrated that P-SiaFNEtoc significantly alleviated bone destruction in the trabecular bone of tibias and increased trabecular bone volume (BV/TV, p = 0.0211) and trabecular number (Tb. N, p = 0.035) (n = 9, unpaired t test, ∗p < 0.05). Representative images are shown. Scale bar is 200 μm.

Article Snippet: Tumours were monitored over 15 days using in vivo bioluminescence imaging. ( g , h ) Pre-treatment of RM1 cells with P-SiaFNEtoc (to remove sialylated glycans) significantly reduced the number of skeletal tumours formed (Mann–Whitney test, p = 0.0454), the incidence of tumour in left tibias (Chi-square test, p = 0.0455), and significantly increased survival time in mice (Log-rank test, p = 0.012). ( i ) Micro-CT analysis demonstrated that P-SiaFNEtoc significantly alleviated bone destruction in the trabecular bone of tibias and increased trabecular bone volume (BV/TV, p = 0.0211) and trabecular number (Tb.

Techniques: Inhibition, Flow Cytometry, Control, Binding Assay, Luciferase, Injection, In Vivo, Imaging, MANN-WHITNEY, Micro-CT

Journal: BMC Cancer

Article Title: Immediate in vivo target-specific cancer cell death after near infrared photoimmunotherapy

doi: 10.1186/1471-2407-12-345

Figure Lengend Snippet: Effect of phototoxicity in response to Pan-IR700 mediated PIT. Cell viability was analyzed with ( A ) bioluminescence and ( B ) fluorescence LIVE/DEAD cytotoxicity assay after NIR light irradiation in MDA-MB-468luc cells.

Article Snippet: For luciferase activity assay, D-luciferin (Gold Biotechnology, St. Louis, MO) was added to culture media at 150 μg/ml and analyzed on a bioluminescence imaging system (Photon Imager; Biospace Lab, Paris, France).

Techniques: Fluorescence, Cytotoxicity Assay, Irradiation

Journal: BMC Cancer

Article Title: Immediate in vivo target-specific cancer cell death after near infrared photoimmunotherapy

doi: 10.1186/1471-2407-12-345

Figure Lengend Snippet: In vivo fluorescence and bioluminescence imaging of orthotopic breast tumors in response to Pan-IR700 mediated PIT. ( A ) Monitoring bioluminescence activity in response to Pan-IR700 mediated PIT ( n = 5 each). B: just before NIR light treatment. Bioluminescence signals of tumors in 100 and 150 J/cm 2 treated groups showed significant difference with that in control or 50 J/cm 2 treated group at all time points examined after PIT ( P <0.001). ( B ) Monitoring bioluminescence activity ratio of treated tumor to internal control ( n = 5 each). B: just before NIR light treatment. Bioluminescence signals of tumors in 100 and 150 J/cm 2 treated groups showed significant difference with that in control or 50 J/cm 2 treated group at all time points examined after PIT ( P <0.001). ( C ) Rapid changes of bioluminescence activity in response to Pan-IR700 mediated PIT ( n = 5 each). Before: 1 d before NIR light treatment. ( D ) IR700 fluorescence and bioluminescence imaging in response to Pan-IR700 mediated PIT. Representative images are shown. Before: just before NIR light treatment, FLI: IR700 fluorescence imaging, WLI: white light imaging, BLI: bioluminescence imaging.

Article Snippet: For luciferase activity assay, D-luciferin (Gold Biotechnology, St. Louis, MO) was added to culture media at 150 μg/ml and analyzed on a bioluminescence imaging system (Photon Imager; Biospace Lab, Paris, France).

Techniques: In Vivo, Fluorescence, Imaging, Activity Assay, Control