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Dual-functional c(RGDyK)-decorated Pluronic micelles designed for antiangiogenesis and the treatment of drug-resistant tumor.

Chen Y, Zhang W, Huang Y, Gao F, Fang X - Int J Nanomedicine (2015)

Bottom Line: Importantly, in vitro antiangiogenesis results demonstrated that c(RGDyK)-FP-DP had a significant inhibition effect on the tubular formation of human umbilical vein endothelial cells and promoted cellular apoptotic activity in MDR KBv cells.In addition, the growth inhibition efficacy of KBv tumor spheroids after crossing the blood-tumor barrier was obviously increased by c(RGDyK)-FP-DP compared to other control groups.Results suggested that c(RGDyK)-decorated Pluronic polymeric micelles can take pharmacological action on both human umbilical vein endothelial cells and KBv MDR cancer cells, resulting in a dual-functional anticancer effect similar to that observed in our in vitro cellular studies.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutics, School of Pharmacy, East China University of Science and Technology, Fudan University, Shanghai, People's Republic of China ; Key Laboratory of Smart Drug Delivery, Ministry of Education and PLA, School of Pharmacy, Fudan University, Shanghai, People's Republic of China.

ABSTRACT
Dual-functional drug delivery system was developed by decorating c(RGDyK) (cyclic RGD [arginine-glycine-aspartic acid] peptide) with Pluronic polymeric micelles (c[RGDyK]-FP-DP) to overcome the drawbacks of low transport of chemotherapeutics across the blood-tumor barrier and poor multidrug-resistant (MDR) tumor therapy. c(RGDyK) that can bind to the integrin protein richly expressed at the site of tumor vascular endothelial cells and tumor cells with high affinity and specificity was conjugated to the N-hydroxysuccinimide-activated PEO terminus of the Pluronic F127 block copolymer. In this study, decreased tumor angiogenic and increased apoptotic activity in MDR cancer cells were observed after the treatment with c(RGDyK)-FP-DP. c(RGDyK)-FP-DP was fully characterized in terms of morphology, particle size, zeta potential, and drug release. Importantly, in vitro antiangiogenesis results demonstrated that c(RGDyK)-FP-DP had a significant inhibition effect on the tubular formation of human umbilical vein endothelial cells and promoted cellular apoptotic activity in MDR KBv cells. In addition, the growth inhibition efficacy of KBv tumor spheroids after crossing the blood-tumor barrier was obviously increased by c(RGDyK)-FP-DP compared to other control groups. Results suggested that c(RGDyK)-decorated Pluronic polymeric micelles can take pharmacological action on both human umbilical vein endothelial cells and KBv MDR cancer cells, resulting in a dual-functional anticancer effect similar to that observed in our in vitro cellular studies.

No MeSH data available.


Related in: MedlinePlus

The qualitative and quantitative analysis for HUVEC cellular uptake.Notes: (A) Qualitative analysis for HUVEC cellular uptake of PF-DOX and c(RGDyK)-FP-DOX at various concentrations; (B) under different incubation times; (C) when preincubated with free c(RGDyK) or incubated at 4°C; (D) quantitative analysis for HUVEC uptake when treated with 50–1,500 μg/mL c(RGDyK)-FP-DOX and PF-DOX at 37°C or 4°C for 60 minutes, respectively; (E) HUVEC uptake when exposed to 100 μg/mL c(RGDyK)-FP-DP and PF-DP at 37°C for different times. Mean ± SD (n=3). Cells were examined by fluorescent microscopy; Red: DOX; Bar: 30 μm.Abbreviations: HUVEC, human umbilical vein endothelial cells; PF-DOX, Pluronic micelles loaded with DOX; c(RGDyK), cyclic RGD peptide; RGD, arginine-glycine-aspartic acid; c(RGDyK)-FP-DOX, c(RGDyK)-decorated Pluronic micelles loaded with DOX; c(RGDyK)-FP-DP, c(RGDyK)-decorated Pluronic micelles loaded with DOX and PTX; PF-DP, Pluronic micelles loaded with DOX and PTX; DOX, doxorubicin; PTX, paclitaxel.
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f4-ijn-10-4863: The qualitative and quantitative analysis for HUVEC cellular uptake.Notes: (A) Qualitative analysis for HUVEC cellular uptake of PF-DOX and c(RGDyK)-FP-DOX at various concentrations; (B) under different incubation times; (C) when preincubated with free c(RGDyK) or incubated at 4°C; (D) quantitative analysis for HUVEC uptake when treated with 50–1,500 μg/mL c(RGDyK)-FP-DOX and PF-DOX at 37°C or 4°C for 60 minutes, respectively; (E) HUVEC uptake when exposed to 100 μg/mL c(RGDyK)-FP-DP and PF-DP at 37°C for different times. Mean ± SD (n=3). Cells were examined by fluorescent microscopy; Red: DOX; Bar: 30 μm.Abbreviations: HUVEC, human umbilical vein endothelial cells; PF-DOX, Pluronic micelles loaded with DOX; c(RGDyK), cyclic RGD peptide; RGD, arginine-glycine-aspartic acid; c(RGDyK)-FP-DOX, c(RGDyK)-decorated Pluronic micelles loaded with DOX; c(RGDyK)-FP-DP, c(RGDyK)-decorated Pluronic micelles loaded with DOX and PTX; PF-DP, Pluronic micelles loaded with DOX and PTX; DOX, doxorubicin; PTX, paclitaxel.

Mentions: DOX was used as a fluorescence probe to label blank micelles for cellular uptake test, the results of which were shown qualitatively by fluorescent images. HUVEC cells treated with either PF-DOX or c(RGDyK)-FP-DOX exhibited variable fluorescent intensity depending on the concentration of micelles (Figure 4A) and the incubation time (Figure 4B). The cellular uptake of c(RGDyK)-FP-DOX in HUVEC cells exhibited concentration- and time-dependent mode, and was higher than those of FP-DOX when the concentration of micelles ranged from 50 μg/mL to 200 μg/mL or the incubation time ranged from 0.5 hour to –2 hours at an identical micelle concentration. Cellular uptake of c(RGDyK)-FP-DOX at 4°C by HUVEC cells was much lower when compared to that at 37°C (Figure 4C). In addition, it was found that after adding free c(RGDyK) peptide, cellular uptake of c(RGDyK)-FP-DOX was reduced (data not shown). Therefore, the cellular uptake of c(RGDyK)-FP-DOX exhibited a concentration-, time- and energy-dependent mode. On preconditioning with c(RGDyK) peptide, ligands of integrin, the cellular uptake of c(RGDyK)-FP-DP was reduced, probably due to the fact that integrin can competitively bind with the free ligands.


Dual-functional c(RGDyK)-decorated Pluronic micelles designed for antiangiogenesis and the treatment of drug-resistant tumor.

Chen Y, Zhang W, Huang Y, Gao F, Fang X - Int J Nanomedicine (2015)

The qualitative and quantitative analysis for HUVEC cellular uptake.Notes: (A) Qualitative analysis for HUVEC cellular uptake of PF-DOX and c(RGDyK)-FP-DOX at various concentrations; (B) under different incubation times; (C) when preincubated with free c(RGDyK) or incubated at 4°C; (D) quantitative analysis for HUVEC uptake when treated with 50–1,500 μg/mL c(RGDyK)-FP-DOX and PF-DOX at 37°C or 4°C for 60 minutes, respectively; (E) HUVEC uptake when exposed to 100 μg/mL c(RGDyK)-FP-DP and PF-DP at 37°C for different times. Mean ± SD (n=3). Cells were examined by fluorescent microscopy; Red: DOX; Bar: 30 μm.Abbreviations: HUVEC, human umbilical vein endothelial cells; PF-DOX, Pluronic micelles loaded with DOX; c(RGDyK), cyclic RGD peptide; RGD, arginine-glycine-aspartic acid; c(RGDyK)-FP-DOX, c(RGDyK)-decorated Pluronic micelles loaded with DOX; c(RGDyK)-FP-DP, c(RGDyK)-decorated Pluronic micelles loaded with DOX and PTX; PF-DP, Pluronic micelles loaded with DOX and PTX; DOX, doxorubicin; PTX, paclitaxel.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4525800&req=5

f4-ijn-10-4863: The qualitative and quantitative analysis for HUVEC cellular uptake.Notes: (A) Qualitative analysis for HUVEC cellular uptake of PF-DOX and c(RGDyK)-FP-DOX at various concentrations; (B) under different incubation times; (C) when preincubated with free c(RGDyK) or incubated at 4°C; (D) quantitative analysis for HUVEC uptake when treated with 50–1,500 μg/mL c(RGDyK)-FP-DOX and PF-DOX at 37°C or 4°C for 60 minutes, respectively; (E) HUVEC uptake when exposed to 100 μg/mL c(RGDyK)-FP-DP and PF-DP at 37°C for different times. Mean ± SD (n=3). Cells were examined by fluorescent microscopy; Red: DOX; Bar: 30 μm.Abbreviations: HUVEC, human umbilical vein endothelial cells; PF-DOX, Pluronic micelles loaded with DOX; c(RGDyK), cyclic RGD peptide; RGD, arginine-glycine-aspartic acid; c(RGDyK)-FP-DOX, c(RGDyK)-decorated Pluronic micelles loaded with DOX; c(RGDyK)-FP-DP, c(RGDyK)-decorated Pluronic micelles loaded with DOX and PTX; PF-DP, Pluronic micelles loaded with DOX and PTX; DOX, doxorubicin; PTX, paclitaxel.
Mentions: DOX was used as a fluorescence probe to label blank micelles for cellular uptake test, the results of which were shown qualitatively by fluorescent images. HUVEC cells treated with either PF-DOX or c(RGDyK)-FP-DOX exhibited variable fluorescent intensity depending on the concentration of micelles (Figure 4A) and the incubation time (Figure 4B). The cellular uptake of c(RGDyK)-FP-DOX in HUVEC cells exhibited concentration- and time-dependent mode, and was higher than those of FP-DOX when the concentration of micelles ranged from 50 μg/mL to 200 μg/mL or the incubation time ranged from 0.5 hour to –2 hours at an identical micelle concentration. Cellular uptake of c(RGDyK)-FP-DOX at 4°C by HUVEC cells was much lower when compared to that at 37°C (Figure 4C). In addition, it was found that after adding free c(RGDyK) peptide, cellular uptake of c(RGDyK)-FP-DOX was reduced (data not shown). Therefore, the cellular uptake of c(RGDyK)-FP-DOX exhibited a concentration-, time- and energy-dependent mode. On preconditioning with c(RGDyK) peptide, ligands of integrin, the cellular uptake of c(RGDyK)-FP-DP was reduced, probably due to the fact that integrin can competitively bind with the free ligands.

Bottom Line: Importantly, in vitro antiangiogenesis results demonstrated that c(RGDyK)-FP-DP had a significant inhibition effect on the tubular formation of human umbilical vein endothelial cells and promoted cellular apoptotic activity in MDR KBv cells.In addition, the growth inhibition efficacy of KBv tumor spheroids after crossing the blood-tumor barrier was obviously increased by c(RGDyK)-FP-DP compared to other control groups.Results suggested that c(RGDyK)-decorated Pluronic polymeric micelles can take pharmacological action on both human umbilical vein endothelial cells and KBv MDR cancer cells, resulting in a dual-functional anticancer effect similar to that observed in our in vitro cellular studies.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutics, School of Pharmacy, East China University of Science and Technology, Fudan University, Shanghai, People's Republic of China ; Key Laboratory of Smart Drug Delivery, Ministry of Education and PLA, School of Pharmacy, Fudan University, Shanghai, People's Republic of China.

ABSTRACT
Dual-functional drug delivery system was developed by decorating c(RGDyK) (cyclic RGD [arginine-glycine-aspartic acid] peptide) with Pluronic polymeric micelles (c[RGDyK]-FP-DP) to overcome the drawbacks of low transport of chemotherapeutics across the blood-tumor barrier and poor multidrug-resistant (MDR) tumor therapy. c(RGDyK) that can bind to the integrin protein richly expressed at the site of tumor vascular endothelial cells and tumor cells with high affinity and specificity was conjugated to the N-hydroxysuccinimide-activated PEO terminus of the Pluronic F127 block copolymer. In this study, decreased tumor angiogenic and increased apoptotic activity in MDR cancer cells were observed after the treatment with c(RGDyK)-FP-DP. c(RGDyK)-FP-DP was fully characterized in terms of morphology, particle size, zeta potential, and drug release. Importantly, in vitro antiangiogenesis results demonstrated that c(RGDyK)-FP-DP had a significant inhibition effect on the tubular formation of human umbilical vein endothelial cells and promoted cellular apoptotic activity in MDR KBv cells. In addition, the growth inhibition efficacy of KBv tumor spheroids after crossing the blood-tumor barrier was obviously increased by c(RGDyK)-FP-DP compared to other control groups. Results suggested that c(RGDyK)-decorated Pluronic polymeric micelles can take pharmacological action on both human umbilical vein endothelial cells and KBv MDR cancer cells, resulting in a dual-functional anticancer effect similar to that observed in our in vitro cellular studies.

No MeSH data available.


Related in: MedlinePlus