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CaCO₃/CaIP₆ composite nanoparticles effectively deliver AKT1 small interfering RNA to inhibit human breast cancer growth.

Zhou H, Wei J, Dai Q, Wang L, Luo J, Cheang T, Wang S - Int J Nanomedicine (2015)

Bottom Line: ACC/CaIP6 nanoparticles effectively transfected cells with little or no toxicity.AKT1 knockdown by ACC/CaIP6/siAKT1 inhibited cell cycle progression and promoted apoptosis of MCF-7 cells.ACC/CaIP6 nanoparticles are a safe and efficient method of delivering siRNA for gene therapy in breast cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurological Intensive Care Unit, First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, People's Republic of China.

ABSTRACT

Background: Small interfering RNA (siRNA)-mediated gene therapy is a promising strategy to temporarily inhibit the expression of genes involved in development of breast cancer. The lack of a safe and efficient gene delivery system has become a major hurdle for siRNA-mediated gene therapy in breast cancer. Our previous studies have demonstrated that inorganic amorphous calcium carbonate (ACC) hybrid nanospheres functionalized with CaIP6 (ACC/CaIP6) nanoparticles are an efficient nucleic acid delivery tool. The present study aimed to evaluate the safety and efficiency of ACC/CaIP6 in delivering siRNA targeting AKT1 (siAKT1) for the treatment of breast cancer.

Methods: The cytotoxicity of the ACC/CaIP6 nanoparticles was evaluated using a tetrazolium assay. The transfection efficiency and intracellular distribution of ACC/siAKT1 were analyzed by flow cytometry and confocal laser scanning microscopy, respectively. A series of in vitro and in vivo assays was performed to evaluate the effects of ACC/CaIP6/siAKT1 on growth of breast cancer cells.

Results: ACC/CaIP6 nanoparticles effectively transfected cells with little or no toxicity. AKT1 knockdown by ACC/CaIP6/siAKT1 inhibited cell cycle progression and promoted apoptosis of MCF-7 cells. Intratumoral injection of ACC/CaIP6/siAKT1 significantly suppressed the growth of breast cancer in mice.

Conclusion: ACC/CaIP6 nanoparticles are a safe and efficient method of delivering siRNA for gene therapy in breast cancer.

No MeSH data available.


Related in: MedlinePlus

ACC/CaIP6 nanoparticle siRNA-binding efficiency and cytotoxicity in MCF-7 cells.Notes: (A) Gel electrophoresis was used to assess ACC/CaIP6 nanoparticles and siRNA binding at different mass ratios. (B) Cytotoxicity was measured by the percentage of viable MCF-7 cells present after ACC/CaIP6 treatment, using the MTT assay 48 hours post–transfection. These percentages were calculated in relation to those of untreated controls. The data show the mean ± standard deviation of three independent experiments (*P<0.05 by one-way analysis of variance).Abbreviations: ACC/CaIP6, amorphous calcium carbonate hybrid nanospheres functionalized with a Ca(II)-inositol hexakisphosphate compound; siNC, control small interfering RNA; lipo, Lipofectamine 2000; siRNA, small interfering RNA.
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f2-ijn-10-4255: ACC/CaIP6 nanoparticle siRNA-binding efficiency and cytotoxicity in MCF-7 cells.Notes: (A) Gel electrophoresis was used to assess ACC/CaIP6 nanoparticles and siRNA binding at different mass ratios. (B) Cytotoxicity was measured by the percentage of viable MCF-7 cells present after ACC/CaIP6 treatment, using the MTT assay 48 hours post–transfection. These percentages were calculated in relation to those of untreated controls. The data show the mean ± standard deviation of three independent experiments (*P<0.05 by one-way analysis of variance).Abbreviations: ACC/CaIP6, amorphous calcium carbonate hybrid nanospheres functionalized with a Ca(II)-inositol hexakisphosphate compound; siNC, control small interfering RNA; lipo, Lipofectamine 2000; siRNA, small interfering RNA.

Mentions: A gel electrophoresis shift assay was performed to confirm ACC/CaIP6/siRNA complexation (Figure 2A). When ACC/CaIP6 and siRNA were present at a mass ratio of 50:1 or greater, the siRNA was completely incorporated into the ACC/CaIP6 nanoparticle complex. These results suggest that the optimal mass ratio for ACC/CaIP6 and siRNA was 50–100 nanoparticles per siRNA (ie, 50–100:1). This is consistent with our previous findings.11,12


CaCO₃/CaIP₆ composite nanoparticles effectively deliver AKT1 small interfering RNA to inhibit human breast cancer growth.

Zhou H, Wei J, Dai Q, Wang L, Luo J, Cheang T, Wang S - Int J Nanomedicine (2015)

ACC/CaIP6 nanoparticle siRNA-binding efficiency and cytotoxicity in MCF-7 cells.Notes: (A) Gel electrophoresis was used to assess ACC/CaIP6 nanoparticles and siRNA binding at different mass ratios. (B) Cytotoxicity was measured by the percentage of viable MCF-7 cells present after ACC/CaIP6 treatment, using the MTT assay 48 hours post–transfection. These percentages were calculated in relation to those of untreated controls. The data show the mean ± standard deviation of three independent experiments (*P<0.05 by one-way analysis of variance).Abbreviations: ACC/CaIP6, amorphous calcium carbonate hybrid nanospheres functionalized with a Ca(II)-inositol hexakisphosphate compound; siNC, control small interfering RNA; lipo, Lipofectamine 2000; siRNA, small interfering RNA.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494612&req=5

f2-ijn-10-4255: ACC/CaIP6 nanoparticle siRNA-binding efficiency and cytotoxicity in MCF-7 cells.Notes: (A) Gel electrophoresis was used to assess ACC/CaIP6 nanoparticles and siRNA binding at different mass ratios. (B) Cytotoxicity was measured by the percentage of viable MCF-7 cells present after ACC/CaIP6 treatment, using the MTT assay 48 hours post–transfection. These percentages were calculated in relation to those of untreated controls. The data show the mean ± standard deviation of three independent experiments (*P<0.05 by one-way analysis of variance).Abbreviations: ACC/CaIP6, amorphous calcium carbonate hybrid nanospheres functionalized with a Ca(II)-inositol hexakisphosphate compound; siNC, control small interfering RNA; lipo, Lipofectamine 2000; siRNA, small interfering RNA.
Mentions: A gel electrophoresis shift assay was performed to confirm ACC/CaIP6/siRNA complexation (Figure 2A). When ACC/CaIP6 and siRNA were present at a mass ratio of 50:1 or greater, the siRNA was completely incorporated into the ACC/CaIP6 nanoparticle complex. These results suggest that the optimal mass ratio for ACC/CaIP6 and siRNA was 50–100 nanoparticles per siRNA (ie, 50–100:1). This is consistent with our previous findings.11,12

Bottom Line: ACC/CaIP6 nanoparticles effectively transfected cells with little or no toxicity.AKT1 knockdown by ACC/CaIP6/siAKT1 inhibited cell cycle progression and promoted apoptosis of MCF-7 cells.ACC/CaIP6 nanoparticles are a safe and efficient method of delivering siRNA for gene therapy in breast cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurological Intensive Care Unit, First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, People's Republic of China.

ABSTRACT

Background: Small interfering RNA (siRNA)-mediated gene therapy is a promising strategy to temporarily inhibit the expression of genes involved in development of breast cancer. The lack of a safe and efficient gene delivery system has become a major hurdle for siRNA-mediated gene therapy in breast cancer. Our previous studies have demonstrated that inorganic amorphous calcium carbonate (ACC) hybrid nanospheres functionalized with CaIP6 (ACC/CaIP6) nanoparticles are an efficient nucleic acid delivery tool. The present study aimed to evaluate the safety and efficiency of ACC/CaIP6 in delivering siRNA targeting AKT1 (siAKT1) for the treatment of breast cancer.

Methods: The cytotoxicity of the ACC/CaIP6 nanoparticles was evaluated using a tetrazolium assay. The transfection efficiency and intracellular distribution of ACC/siAKT1 were analyzed by flow cytometry and confocal laser scanning microscopy, respectively. A series of in vitro and in vivo assays was performed to evaluate the effects of ACC/CaIP6/siAKT1 on growth of breast cancer cells.

Results: ACC/CaIP6 nanoparticles effectively transfected cells with little or no toxicity. AKT1 knockdown by ACC/CaIP6/siAKT1 inhibited cell cycle progression and promoted apoptosis of MCF-7 cells. Intratumoral injection of ACC/CaIP6/siAKT1 significantly suppressed the growth of breast cancer in mice.

Conclusion: ACC/CaIP6 nanoparticles are a safe and efficient method of delivering siRNA for gene therapy in breast cancer.

No MeSH data available.


Related in: MedlinePlus