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Development of in vitro gene delivery system using ORMOSIL nanoparticle: Analysis of p53 gene expression in cultured breast cancer cell (MCF-7).

Rejeeth C, Kannan S, Muthuchelian K - Cancer Nanotechnol (2012)

Bottom Line: Interesting agarose gel electrophoresis studies revealed that the nanoparticles efficiently complex with pCMV-Myc vector.Whereas, the growth rate was significantly reduced in ORMOSIL/p53/pCMV-Myc transfected breast cancer cells compared to the growth rate of non-transfected cells.The results of this approach using ORMOSIL nanoparticles as a non-viral gene delivery platform have a promising future for use as effective transfection agent for therapeutic manipulation of cancer cells and targeted cancer gene therapy in vivo.

View Article: PubMed Central - PubMed

Affiliation: Proteomics and Molecular Cell Physiology Lab, Department of Zoology, School of Life Sciences, Bharathiar University, Coimbatore, 641 046, TN India.

ABSTRACT

This article reports on the application of organically modified silica (ORMOSIL) nanoparticles as an efficient in vitro gene delivery system in the recent years. Based on that prime objective, the present study addresses the possible ways to reduce cancers incidence at cellular level. In this context, ORMOSIL nanoparticles had been synthesized and incubated along with pCMV-Myc (3.8 kb) plasmid vector construct carrying p53gene, and transfected into the breast cancer cell line MCF-7 cells. Western blot analysis showed that the p53 protein was significantly expressed in breast cancer cell upon transfection. The confocal and electron microscopic studies further confirmed that the nanoparticles were accumulated in the cytoplasm and the nucleus of the cancer cells transfected with p53 gene. Interesting agarose gel electrophoresis studies revealed that the nanoparticles efficiently complex with pCMV-Myc vector. The anti-cancer properties of p53 were demonstrated by assessing the cell survival and growth rate which showed a positive linear correlation in cancer cells. Whereas, the growth rate was significantly reduced in ORMOSIL/p53/pCMV-Myc transfected breast cancer cells compared to the growth rate of non-transfected cells. The results of this approach using ORMOSIL nanoparticles as a non-viral gene delivery platform have a promising future for use as effective transfection agent for therapeutic manipulation of cancer cells and targeted cancer gene therapy in vivo.

No MeSH data available.


Related in: MedlinePlus

Transmission electron microscopy images of highly monodispersed ORMOSIL nanoparticles
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Fig1: Transmission electron microscopy images of highly monodispersed ORMOSIL nanoparticles

Mentions: Figure 1 shows transmission electron microscopy images of ORMOSIL nanoparticles. Note that the particles are spherical in shape and highly monodispersed. For the preparation of cationic ORMOSILNs, VTES was selected as a silica component that would constitute the core of the ORMOSILNs. Aerosol-OT, DMSO, and APTES were used as cationic ORMOSIL, helper VTES, and surfactant, respectively. The mean diameter of the ORMOSILNs was under 60 nm, which is generally known to be an effective size for high transfection efficiency. The zeta potentials of ORMOSILNs and ORMOSILNs/DNA were approximately 8 and 13 mV, respectively. To investigate whether the changes in the packing material or storage conditions may further improve the stability of freeze-dried ORMOSILNs, ORMOSILNs were freeze-dried using sucrose as a cryoprotectant, and the particle diameter and transfection efficiency were subsequently determined. The particle diameters of ORMOSILNs and ORMOSILNs/DNA were slightly increased but both of them were under 60 nm. We also compared the transfection efficiencies of the ORMOSILNs before freeze drying (Table 1).Fig. 1


Development of in vitro gene delivery system using ORMOSIL nanoparticle: Analysis of p53 gene expression in cultured breast cancer cell (MCF-7).

Rejeeth C, Kannan S, Muthuchelian K - Cancer Nanotechnol (2012)

Transmission electron microscopy images of highly monodispersed ORMOSIL nanoparticles
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4544070&req=5

Fig1: Transmission electron microscopy images of highly monodispersed ORMOSIL nanoparticles
Mentions: Figure 1 shows transmission electron microscopy images of ORMOSIL nanoparticles. Note that the particles are spherical in shape and highly monodispersed. For the preparation of cationic ORMOSILNs, VTES was selected as a silica component that would constitute the core of the ORMOSILNs. Aerosol-OT, DMSO, and APTES were used as cationic ORMOSIL, helper VTES, and surfactant, respectively. The mean diameter of the ORMOSILNs was under 60 nm, which is generally known to be an effective size for high transfection efficiency. The zeta potentials of ORMOSILNs and ORMOSILNs/DNA were approximately 8 and 13 mV, respectively. To investigate whether the changes in the packing material or storage conditions may further improve the stability of freeze-dried ORMOSILNs, ORMOSILNs were freeze-dried using sucrose as a cryoprotectant, and the particle diameter and transfection efficiency were subsequently determined. The particle diameters of ORMOSILNs and ORMOSILNs/DNA were slightly increased but both of them were under 60 nm. We also compared the transfection efficiencies of the ORMOSILNs before freeze drying (Table 1).Fig. 1

Bottom Line: Interesting agarose gel electrophoresis studies revealed that the nanoparticles efficiently complex with pCMV-Myc vector.Whereas, the growth rate was significantly reduced in ORMOSIL/p53/pCMV-Myc transfected breast cancer cells compared to the growth rate of non-transfected cells.The results of this approach using ORMOSIL nanoparticles as a non-viral gene delivery platform have a promising future for use as effective transfection agent for therapeutic manipulation of cancer cells and targeted cancer gene therapy in vivo.

View Article: PubMed Central - PubMed

Affiliation: Proteomics and Molecular Cell Physiology Lab, Department of Zoology, School of Life Sciences, Bharathiar University, Coimbatore, 641 046, TN India.

ABSTRACT

This article reports on the application of organically modified silica (ORMOSIL) nanoparticles as an efficient in vitro gene delivery system in the recent years. Based on that prime objective, the present study addresses the possible ways to reduce cancers incidence at cellular level. In this context, ORMOSIL nanoparticles had been synthesized and incubated along with pCMV-Myc (3.8 kb) plasmid vector construct carrying p53gene, and transfected into the breast cancer cell line MCF-7 cells. Western blot analysis showed that the p53 protein was significantly expressed in breast cancer cell upon transfection. The confocal and electron microscopic studies further confirmed that the nanoparticles were accumulated in the cytoplasm and the nucleus of the cancer cells transfected with p53 gene. Interesting agarose gel electrophoresis studies revealed that the nanoparticles efficiently complex with pCMV-Myc vector. The anti-cancer properties of p53 were demonstrated by assessing the cell survival and growth rate which showed a positive linear correlation in cancer cells. Whereas, the growth rate was significantly reduced in ORMOSIL/p53/pCMV-Myc transfected breast cancer cells compared to the growth rate of non-transfected cells. The results of this approach using ORMOSIL nanoparticles as a non-viral gene delivery platform have a promising future for use as effective transfection agent for therapeutic manipulation of cancer cells and targeted cancer gene therapy in vivo.

No MeSH data available.


Related in: MedlinePlus