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Coating Solid Lipid Nanoparticles with Hyaluronic Acid Enhances Antitumor Activity against Melanoma Stem-like Cells.

Shen H, Shi S, Zhang Z, Gong T, Sun X - Theranostics (2015)

Bottom Line: First, we developed a model system based on melanoma stem-like cells for experiments in vitro and in mouse xenografts, and we showed that cells expressing high levels of CD44 (CD44(+)) displayed a strong CSC phenotype while cells expressing low levels of CD44 (CD44(-)) did not.In the B16F10-CD44(+) lung metastasis model, PTX-loaded HA-SLNs targeted the tumor-bearing lung tissues well and subsequently exhibited significant antitumor effects with a relative low dose of PTX, which provided significant survival benefit without evidence of adverse events.These findings suggest that the HA-SLNs targeting system shows promise for enhancing cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Drug Targeting and Drug Delivery Systems, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu 610041, P. R. China.

ABSTRACT
Successful anticancer chemotherapy requires targeting tumors efficiently and further potential to eliminate cancer stem cell (CSC) subpopulations. Since CD44 is present on many types of CSCs, and it binds specially to hyaluronic acid (HA), we tested whether coating solid lipid nanoparticles with hyaluronan (HA-SLNs)would allow targeted delivery of paclitaxel (PTX) to CD44-overexpressing B16F10 melanoma cells. First, we developed a model system based on melanoma stem-like cells for experiments in vitro and in mouse xenografts, and we showed that cells expressing high levels of CD44 (CD44(+)) displayed a strong CSC phenotype while cells expressing low levels of CD44 (CD44(-)) did not. This phenotype included sphere and colony formation, higher proportion of side population cells, expression of CSC-related markers (ALDH, CD133, Oct-4) and tumorigenicity in vivo. Next we showed that administering PTX-loaded HA-SLNs led to efficient intracellular delivery of PTX and induced substantial apoptosis in CD44(+) cells in vitro. In the B16F10-CD44(+) lung metastasis model, PTX-loaded HA-SLNs targeted the tumor-bearing lung tissues well and subsequently exhibited significant antitumor effects with a relative low dose of PTX, which provided significant survival benefit without evidence of adverse events. These findings suggest that the HA-SLNs targeting system shows promise for enhancing cancer therapy.

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CSC phenotype of B16F10-CD44+ cells. A) Sphere formation and soft agar colony formation assays using B16F10-CD44+ and B16F10-CD44- cells. B) Quantitative RT-PCR determination of ALDH and CD133 expression. Data were shown as normalized fold expression relative to β-actin as an internal control (n = 3). C) Representative results of a spheres formation assay in which B16F10-CD44+ cell were cultured for 2-3 weeks, treated for 4 h with HA-SLNs/PTX and photographed 48 h later. D) Representative tumors and tumor volume in C57BL/6 mice injected subcutaneously with B16F10-CD44+ or B16F10-CD44- cells (5 × 104 cells/animal), and tumor growth was monitored for 23 days after injection. E) Identification of the side population (SP) and the proportion of SP cells in B16F10-CD44- cells and in B16F10-CD44+ cells (untreated or treated with SLNs/PTX or HA-SLNs/PTX). As a control for identify of SP cells, experiments were also performed in the presence of the ABC transporter inhibitor verapamil. Data were shown as mean ± SD (n = 3).
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Figure 5: CSC phenotype of B16F10-CD44+ cells. A) Sphere formation and soft agar colony formation assays using B16F10-CD44+ and B16F10-CD44- cells. B) Quantitative RT-PCR determination of ALDH and CD133 expression. Data were shown as normalized fold expression relative to β-actin as an internal control (n = 3). C) Representative results of a spheres formation assay in which B16F10-CD44+ cell were cultured for 2-3 weeks, treated for 4 h with HA-SLNs/PTX and photographed 48 h later. D) Representative tumors and tumor volume in C57BL/6 mice injected subcutaneously with B16F10-CD44+ or B16F10-CD44- cells (5 × 104 cells/animal), and tumor growth was monitored for 23 days after injection. E) Identification of the side population (SP) and the proportion of SP cells in B16F10-CD44- cells and in B16F10-CD44+ cells (untreated or treated with SLNs/PTX or HA-SLNs/PTX). As a control for identify of SP cells, experiments were also performed in the presence of the ABC transporter inhibitor verapamil. Data were shown as mean ± SD (n = 3).

Mentions: B16F10-CD44+ cells grew as spheroids and cell colonies under our experimental conditions, whereas B16F10-CD44- cells did not (Fig. 5A). Quantitative real-time PCR showed that B16F10-CD44+ cells expressed significantly higher levels of ALDH and CD133 (Fig. 5B). When mice were injected subcutaneously with 5 ×104 B16F10-CD44+ or CD44- cells, CD44+ cells formed tumors (average size, 219.55 mm3), whereas CD44- cells did not (Fig. 5D). Taken together, these findings suggest that B16F10-CD44+ cells, but not B16F10-CD44- cells, have substantial expression of CSC marker, self-renewal activity and tumorigenicity in vitro and in vivo, consistent with a CSC phenotype. This implies that PTX-loaded nanoparticles should target B16F10-CD44+ cells in order to provide effective antitumor therapy.


Coating Solid Lipid Nanoparticles with Hyaluronic Acid Enhances Antitumor Activity against Melanoma Stem-like Cells.

Shen H, Shi S, Zhang Z, Gong T, Sun X - Theranostics (2015)

CSC phenotype of B16F10-CD44+ cells. A) Sphere formation and soft agar colony formation assays using B16F10-CD44+ and B16F10-CD44- cells. B) Quantitative RT-PCR determination of ALDH and CD133 expression. Data were shown as normalized fold expression relative to β-actin as an internal control (n = 3). C) Representative results of a spheres formation assay in which B16F10-CD44+ cell were cultured for 2-3 weeks, treated for 4 h with HA-SLNs/PTX and photographed 48 h later. D) Representative tumors and tumor volume in C57BL/6 mice injected subcutaneously with B16F10-CD44+ or B16F10-CD44- cells (5 × 104 cells/animal), and tumor growth was monitored for 23 days after injection. E) Identification of the side population (SP) and the proportion of SP cells in B16F10-CD44- cells and in B16F10-CD44+ cells (untreated or treated with SLNs/PTX or HA-SLNs/PTX). As a control for identify of SP cells, experiments were also performed in the presence of the ABC transporter inhibitor verapamil. Data were shown as mean ± SD (n = 3).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4402499&req=5

Figure 5: CSC phenotype of B16F10-CD44+ cells. A) Sphere formation and soft agar colony formation assays using B16F10-CD44+ and B16F10-CD44- cells. B) Quantitative RT-PCR determination of ALDH and CD133 expression. Data were shown as normalized fold expression relative to β-actin as an internal control (n = 3). C) Representative results of a spheres formation assay in which B16F10-CD44+ cell were cultured for 2-3 weeks, treated for 4 h with HA-SLNs/PTX and photographed 48 h later. D) Representative tumors and tumor volume in C57BL/6 mice injected subcutaneously with B16F10-CD44+ or B16F10-CD44- cells (5 × 104 cells/animal), and tumor growth was monitored for 23 days after injection. E) Identification of the side population (SP) and the proportion of SP cells in B16F10-CD44- cells and in B16F10-CD44+ cells (untreated or treated with SLNs/PTX or HA-SLNs/PTX). As a control for identify of SP cells, experiments were also performed in the presence of the ABC transporter inhibitor verapamil. Data were shown as mean ± SD (n = 3).
Mentions: B16F10-CD44+ cells grew as spheroids and cell colonies under our experimental conditions, whereas B16F10-CD44- cells did not (Fig. 5A). Quantitative real-time PCR showed that B16F10-CD44+ cells expressed significantly higher levels of ALDH and CD133 (Fig. 5B). When mice were injected subcutaneously with 5 ×104 B16F10-CD44+ or CD44- cells, CD44+ cells formed tumors (average size, 219.55 mm3), whereas CD44- cells did not (Fig. 5D). Taken together, these findings suggest that B16F10-CD44+ cells, but not B16F10-CD44- cells, have substantial expression of CSC marker, self-renewal activity and tumorigenicity in vitro and in vivo, consistent with a CSC phenotype. This implies that PTX-loaded nanoparticles should target B16F10-CD44+ cells in order to provide effective antitumor therapy.

Bottom Line: First, we developed a model system based on melanoma stem-like cells for experiments in vitro and in mouse xenografts, and we showed that cells expressing high levels of CD44 (CD44(+)) displayed a strong CSC phenotype while cells expressing low levels of CD44 (CD44(-)) did not.In the B16F10-CD44(+) lung metastasis model, PTX-loaded HA-SLNs targeted the tumor-bearing lung tissues well and subsequently exhibited significant antitumor effects with a relative low dose of PTX, which provided significant survival benefit without evidence of adverse events.These findings suggest that the HA-SLNs targeting system shows promise for enhancing cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Drug Targeting and Drug Delivery Systems, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu 610041, P. R. China.

ABSTRACT
Successful anticancer chemotherapy requires targeting tumors efficiently and further potential to eliminate cancer stem cell (CSC) subpopulations. Since CD44 is present on many types of CSCs, and it binds specially to hyaluronic acid (HA), we tested whether coating solid lipid nanoparticles with hyaluronan (HA-SLNs)would allow targeted delivery of paclitaxel (PTX) to CD44-overexpressing B16F10 melanoma cells. First, we developed a model system based on melanoma stem-like cells for experiments in vitro and in mouse xenografts, and we showed that cells expressing high levels of CD44 (CD44(+)) displayed a strong CSC phenotype while cells expressing low levels of CD44 (CD44(-)) did not. This phenotype included sphere and colony formation, higher proportion of side population cells, expression of CSC-related markers (ALDH, CD133, Oct-4) and tumorigenicity in vivo. Next we showed that administering PTX-loaded HA-SLNs led to efficient intracellular delivery of PTX and induced substantial apoptosis in CD44(+) cells in vitro. In the B16F10-CD44(+) lung metastasis model, PTX-loaded HA-SLNs targeted the tumor-bearing lung tissues well and subsequently exhibited significant antitumor effects with a relative low dose of PTX, which provided significant survival benefit without evidence of adverse events. These findings suggest that the HA-SLNs targeting system shows promise for enhancing cancer therapy.

Show MeSH
Related in: MedlinePlus