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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

FTIR spectra of neat VTES and APTES ORMOSIL nanoparticles
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Fig3: FTIR spectra of neat VTES and APTES ORMOSIL nanoparticles

Mentions: The nanoparticle surface design involves an optimum balance of the use of inert and active surface functional groups to achieve minimal nanoparticle aggregation and reduce nanoparticle non-specific binding. Silica nanoparticles were prepared in a water-in-oil microemulsion and subsequently surface-modified via co-hydrolysis with tetraethylorthosilicate (TEOS) and organosilane reagents. Nanoparticles with different functional groups, including amine and amine/phosphonate, were produced in the present investigation. The FT-IR spectrum (Fig. 3) of synthesized hybrid amino-functionalized ORMOSIL nanoparticles by a synchronous hydrolysis of VTES and 3-aminopropyltriethoxysilane (APTES) sample shows peaks around 1,730 and 3,516 cm−1 corresponding to the carboxyl and hydroxyl groups (Shan et al. 2000). The IR spectrum shows the peaks for NH2 group as well as silica. The amino-functionalized nanoparticles are able to electrostatically condense DNA (both gene of interest and genomic) and protect it from enzymatic degradation. Hence, silica was functionalized with amino group in the present investigation. Furthermore, in the given IR spectrum the amino group peaks were noticed between 3,000 and 3,600 (stretched vibration) and also between 1,550 and 1,640 (bending vibration). Interestingly, the peak for silica is observed between 1,000 and 1,250.Fig. 3


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)

FTIR spectra of neat VTES and APTES ORMOSIL nanoparticles
© Copyright Policy
Related In: Results  -  Collection

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

Fig3: FTIR spectra of neat VTES and APTES ORMOSIL nanoparticles
Mentions: The nanoparticle surface design involves an optimum balance of the use of inert and active surface functional groups to achieve minimal nanoparticle aggregation and reduce nanoparticle non-specific binding. Silica nanoparticles were prepared in a water-in-oil microemulsion and subsequently surface-modified via co-hydrolysis with tetraethylorthosilicate (TEOS) and organosilane reagents. Nanoparticles with different functional groups, including amine and amine/phosphonate, were produced in the present investigation. The FT-IR spectrum (Fig. 3) of synthesized hybrid amino-functionalized ORMOSIL nanoparticles by a synchronous hydrolysis of VTES and 3-aminopropyltriethoxysilane (APTES) sample shows peaks around 1,730 and 3,516 cm−1 corresponding to the carboxyl and hydroxyl groups (Shan et al. 2000). The IR spectrum shows the peaks for NH2 group as well as silica. The amino-functionalized nanoparticles are able to electrostatically condense DNA (both gene of interest and genomic) and protect it from enzymatic degradation. Hence, silica was functionalized with amino group in the present investigation. Furthermore, in the given IR spectrum the amino group peaks were noticed between 3,000 and 3,600 (stretched vibration) and also between 1,550 and 1,640 (bending vibration). Interestingly, the peak for silica is observed between 1,000 and 1,250.Fig. 3

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