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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

In vitro AKT1 silencing by ACC/CaIP6/siAKT1 transfection in MCF-7 cells.Notes: (A) AKT1 mRNA and protein expression after transfecting MCF-7 cells with ACC/CaIP6/siAKT1 complexes was assessed by real-time polymerase chain reaction and Western blots, respectively. ***P<0.001 versus control. (B) The MTT assay was used to assess MCF-7 cell viability after transfecting cells with ACC/CaIP6 complexes containing different concentrations of siAKT1 (*P<0.05, ***P<0.001 versus control). (C) Flow cytometry was used to determine the cell cycles of MCF-7 cells 48 hours after ACC/CaIP6/siAKT1 transfection. (D) Apoptosis was measured using Annexin V-phycoerythrin/propidium iodide staining for MCF-7 cells treated with various ACC/CaIP6/siRNA complexes (one of three replicates is shown).Abbreviations: ACC, amorphous cal cium carbonate; ACC/CaIP6, amorphous calcium carbonate hybrid nanospheres functionalized with a Ca(II)-inositol hexakisphosphate compound; siAKT1, small interfering AKT1; siNC, control small interfering RNA; OD, optical density; PI, propidium iodide.
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f5-ijn-10-4255: In vitro AKT1 silencing by ACC/CaIP6/siAKT1 transfection in MCF-7 cells.Notes: (A) AKT1 mRNA and protein expression after transfecting MCF-7 cells with ACC/CaIP6/siAKT1 complexes was assessed by real-time polymerase chain reaction and Western blots, respectively. ***P<0.001 versus control. (B) The MTT assay was used to assess MCF-7 cell viability after transfecting cells with ACC/CaIP6 complexes containing different concentrations of siAKT1 (*P<0.05, ***P<0.001 versus control). (C) Flow cytometry was used to determine the cell cycles of MCF-7 cells 48 hours after ACC/CaIP6/siAKT1 transfection. (D) Apoptosis was measured using Annexin V-phycoerythrin/propidium iodide staining for MCF-7 cells treated with various ACC/CaIP6/siRNA complexes (one of three replicates is shown).Abbreviations: ACC, amorphous cal cium carbonate; ACC/CaIP6, amorphous calcium carbonate hybrid nanospheres functionalized with a Ca(II)-inositol hexakisphosphate compound; siAKT1, small interfering AKT1; siNC, control small interfering RNA; OD, optical density; PI, propidium iodide.

Mentions: Real-time PCR and Western blots were used to measure AKT1 expression and confirm the knockdown of AKT1 following siAKT1 delivery. ACC/CaIP6/siAKT1 nanoparticles efficiently silenced AKT1 expression at both the mRNA and protein levels in MCF-7 cells in an siRNA dose-dependent manner (Figure 5A). This is consistent with the improved transfection efficiency that resulted from increasing the siRNA concentration (Figure 3B). In contrast, the nonloaded nanoparticles did not decrease AKT1 expression in MCF-7 cells.


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)

In vitro AKT1 silencing by ACC/CaIP6/siAKT1 transfection in MCF-7 cells.Notes: (A) AKT1 mRNA and protein expression after transfecting MCF-7 cells with ACC/CaIP6/siAKT1 complexes was assessed by real-time polymerase chain reaction and Western blots, respectively. ***P<0.001 versus control. (B) The MTT assay was used to assess MCF-7 cell viability after transfecting cells with ACC/CaIP6 complexes containing different concentrations of siAKT1 (*P<0.05, ***P<0.001 versus control). (C) Flow cytometry was used to determine the cell cycles of MCF-7 cells 48 hours after ACC/CaIP6/siAKT1 transfection. (D) Apoptosis was measured using Annexin V-phycoerythrin/propidium iodide staining for MCF-7 cells treated with various ACC/CaIP6/siRNA complexes (one of three replicates is shown).Abbreviations: ACC, amorphous cal cium carbonate; ACC/CaIP6, amorphous calcium carbonate hybrid nanospheres functionalized with a Ca(II)-inositol hexakisphosphate compound; siAKT1, small interfering AKT1; siNC, control small interfering RNA; OD, optical density; PI, propidium iodide.
© Copyright Policy
Related In: Results  -  Collection

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

f5-ijn-10-4255: In vitro AKT1 silencing by ACC/CaIP6/siAKT1 transfection in MCF-7 cells.Notes: (A) AKT1 mRNA and protein expression after transfecting MCF-7 cells with ACC/CaIP6/siAKT1 complexes was assessed by real-time polymerase chain reaction and Western blots, respectively. ***P<0.001 versus control. (B) The MTT assay was used to assess MCF-7 cell viability after transfecting cells with ACC/CaIP6 complexes containing different concentrations of siAKT1 (*P<0.05, ***P<0.001 versus control). (C) Flow cytometry was used to determine the cell cycles of MCF-7 cells 48 hours after ACC/CaIP6/siAKT1 transfection. (D) Apoptosis was measured using Annexin V-phycoerythrin/propidium iodide staining for MCF-7 cells treated with various ACC/CaIP6/siRNA complexes (one of three replicates is shown).Abbreviations: ACC, amorphous cal cium carbonate; ACC/CaIP6, amorphous calcium carbonate hybrid nanospheres functionalized with a Ca(II)-inositol hexakisphosphate compound; siAKT1, small interfering AKT1; siNC, control small interfering RNA; OD, optical density; PI, propidium iodide.
Mentions: Real-time PCR and Western blots were used to measure AKT1 expression and confirm the knockdown of AKT1 following siAKT1 delivery. ACC/CaIP6/siAKT1 nanoparticles efficiently silenced AKT1 expression at both the mRNA and protein levels in MCF-7 cells in an siRNA dose-dependent manner (Figure 5A). This is consistent with the improved transfection efficiency that resulted from increasing the siRNA concentration (Figure 3B). In contrast, the nonloaded nanoparticles did not decrease AKT1 expression in MCF-7 cells.

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