- Title
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Platinum-triggered Bond-cleavage of Pentynoyl amide and N-propargyl handles for Drug-Activation
- Authors
- Oliveira, B.L., Stenton, B.J., V B, U., de Almeida, C.R., Conde, J., Negrão, M., Schneider, F.S.S., Cordeiro, C., Godinho Ferreira, M., Caramori, G.F., Domingos, J.B., Fior, R., Bernardes, G.J.L.
- Source
- Full text @ J. Am. Chem. Soc.
CisPt Decages the Fluorogenic Probe 9 in vivo. Zebrafish larvae were exposed to 9 diluted in embryonic medium for 24 h, followed by a 1 h wash in embryonic medium. Larvae were randomly distributed into two conditions: DMSO or CisPt for 24 h (a). Confocal image of zebrafish larvae exposed to 9 + DMSO (b) and 9 + CisPt (c). |
CisPt-mediated prodrug decaging in zebrafish xenografts. HCT116 human CRC cells were fluorescently labeled with lipophilic CM-DiI (shown in red) and injected into the perivitelline space (PVS) 2 days post fertilization (dpf) Tg(Fli1:eGFP) zebrafish larvae. Zebrafish xenografts were randomly distributed into treatment groups, daily treated with DMSO, CisPt, pFU, and pFU+CisPt and analyzed at 4, 6, or 7dpi for proliferation, apoptosis and tumor size. At 4 dpi, 6 dpi, and 7dpi, zebrafish xenografts were imaged by stereoscope (a–l) and by confocal microscopy (a′–l′DAPI plus DiI, a′′–l′′ maximum projection of activated caspase 3). Proliferation (mitotic figures: m; p, *P = 0.0104, ***P = 0.0004, ****P < 0.0001; s, **P = 0.0023, *** P = 0.0002), apoptosis (activated caspase 3: n, **P = 0.0033, ***P = 0.0006; q, *P = 0.0126, ****P < 0.0001; t, **P = 0.0068) and tumor size (n° of tumor cells: o, *P = 0.0279; r, ****P < 0.0001; u, *P = 0.0411, **P = 0.0010) were analyzed and quantified. Graphs represent fold induction (normalized values to controls) of Avg ± SEM. The number of xenografts analyzed is indicated in the representative images and each dot represents one zebrafish xenograft. Statistical analysis was performed using an unpaired test. Statistical results: ns > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, and ****P ≤ 0.0001. All images are anterior to the left, posterior to right, dorsal up, and ventral down. Scale bar 50 μm. |
Cisplatin-mediated prodrug decaging in a colon cancer Zebrafish Xenograft model at 6dpi. HCT116 human CRC cells were fluorescently labelled with lipophilic dye CM-DiI (shown in red) and injected into the PVS of 2 dpf Tg(Fli1:eGFP) zebrafish larvae. Zebrafish xenografts were treated in vivo for 5 consecutive days with: DMSO, 5-FU, CisPt, cFU, pFU, cFU+CispPt, pFU+CisPt and 5-FU+CisPt. At 6dpi (5dpt), zebrafish xenografts were imaged by stereoscope (A-H) and by confocal microscopy (A’-H’ DAPI plus DiI; A’’-H’’ a maximum projection of activated caspase 3). Quantification of proliferation (mitotic figures, I *** P=0.0004, ** P=0.0012, **** P<0.0001, * P=0.0106, * pFU vs pFU+CisPt P=0.0104, * cFU+CisPt vs pFU+CisPt P=0.0470), apoptosis (activated caspase3, J ****P<0.0001, * P=0.0126, ** P=0.0084, *** P=0.0002) and tumor size (no of tumor cells, K *** P=0.0008, * cFU P=0.0216, **** P<0.0001, * 5-FU+CisPt P=0.0273, ** P=0.0017). Outcomes are expressed by fold induction (normalized values to controls) as AVG ± SEM. The number of xenografts analyzed is indicated in the representative images and each dot represents one zebrafish xenograft. Statistical analysis was performed using an unpaired t-test. Statistical results: ns>0.05, *P≤0.05, **P≤0.01, ***P≤0.001 and ****P≤0.0001. All images are anterior to the left, posterior to right, dorsal up, and ventral down. Scale bar 50 μm. |
Cisplatin-mediated prodrug decaging in a colon cancer Zebrafish Xenograft model at 7dpi. HCT116 human CRC cells were fluorescently labelled with lipophilic dye CM-DiI and injected into the PVS of 2 dpf Tg(Fli1:eGFP) zebrafish larvae. Zebrafish xenografts were treated in vivo for 6 consecutive days with: DMSO, Cispt, cFU, pFU, cFU+CisPt and pFU+CisPt. At 7dpi (6dpt), zebrafish xenografts were imaged by stereoscope (A-F) and by confocal microscopy (A’-F’ DAPI plus DiI, A’’-F’’ a maximum projection of activated caspase 3). Quantification of proliferation (mitotic figures, G * P=0.0180, ** P=0.0023, *** P=0.0002), apoptosis (activated caspase3, H * P=0.0119, ** P=0.0068) and tumor size (no of tumor cells, I ** P=0.0010, * cFU+CisPt vs pFU+CisPt P=0.0186, * pFU vs pFU+CisPt P=0.0411). Outcomes are expressed by fold induction (normalized values to controls) as AVG ± SEM. The number of xenografts analyzed is indicated in the representative images and each dot represents one zebrafish xenograft. Statistical analysis was performed using an unpaired t-test. Statistical results: ns>0.05, *P≤0.05, **P≤0.01, ***P≤0.001 and ****P≤0.0001. All images are anterior to the left, posterior to right, dorsal up, and ventral down. Scale bar 50 μm. |