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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

The in vitro growth inhibition of the MCF-7 cells after transfection with DNA/ORMOSILNs complexes. The number of adherent cells was determined at each time point. Each point indicates the mean ± SD of values obtained from the assay of cells from six replicate microtiter plate wells. *p < 0.05, compared with the in vitro growth inhibition of the control
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Fig7: The in vitro growth inhibition of the MCF-7 cells after transfection with DNA/ORMOSILNs complexes. The number of adherent cells was determined at each time point. Each point indicates the mean ± SD of values obtained from the assay of cells from six replicate microtiter plate wells. *p < 0.05, compared with the in vitro growth inhibition of the control

Mentions: Using in vitro growth assay, the effect of the transfection of p53–pCMV/ORMOSILNs on the rate of MCF-7 cell growth was determined by measuring the doubling time compared to that of uninfected control cells. On day 0, the cell lines were passed into monolayer cultures in 24-well flat-bottomed microplates. The following day, the cells were transfected with plasmid DNA alone, DNA/ORMOSILNs, or DNA/Lipofectin®. The growth rate of p53–pCMV/ORMOSILNs-transfected cells was significantly inhibited compared with control cells or cells transfected with p53 DNA alone (Fig. 7). These results indicate that ORMSILNs-mediated p53 DNA delivery inhibited the growth of MCF-7 cells. Transfection of MCF-7 cells with p53 DNA alone had a slight effect on cell growth compared to the uninfected cells. Thus, the inhibitory p53–pCMV/ORMOSILNs transfection in MCF-7 cells is likely due to the expression of p53 protein.Fig. 7


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)

The in vitro growth inhibition of the MCF-7 cells after transfection with DNA/ORMOSILNs complexes. The number of adherent cells was determined at each time point. Each point indicates the mean ± SD of values obtained from the assay of cells from six replicate microtiter plate wells. *p < 0.05, compared with the in vitro growth inhibition of the control
© Copyright Policy
Related In: Results  -  Collection

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

Fig7: The in vitro growth inhibition of the MCF-7 cells after transfection with DNA/ORMOSILNs complexes. The number of adherent cells was determined at each time point. Each point indicates the mean ± SD of values obtained from the assay of cells from six replicate microtiter plate wells. *p < 0.05, compared with the in vitro growth inhibition of the control
Mentions: Using in vitro growth assay, the effect of the transfection of p53–pCMV/ORMOSILNs on the rate of MCF-7 cell growth was determined by measuring the doubling time compared to that of uninfected control cells. On day 0, the cell lines were passed into monolayer cultures in 24-well flat-bottomed microplates. The following day, the cells were transfected with plasmid DNA alone, DNA/ORMOSILNs, or DNA/Lipofectin®. The growth rate of p53–pCMV/ORMOSILNs-transfected cells was significantly inhibited compared with control cells or cells transfected with p53 DNA alone (Fig. 7). These results indicate that ORMSILNs-mediated p53 DNA delivery inhibited the growth of MCF-7 cells. Transfection of MCF-7 cells with p53 DNA alone had a slight effect on cell growth compared to the uninfected cells. Thus, the inhibitory p53–pCMV/ORMOSILNs transfection in MCF-7 cells is likely due to the expression of p53 protein.Fig. 7

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