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Development of in vitro gene delivery system using ORMOSIL nanoparticle: Analysis of p53 gene expression in cultured
Chandrababu Rejeeth1, Soundarapandian Kannan1, Krishnasamy Muthuchelian2
1Proteomics 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.
Insights
Organically modified silica (ORMOSIL) nanoparticles efficiently deliver the p53 gene into breast cancer cells, significantly reducing their growth. This non-viral gene delivery system shows promise for targeted cancer gene therapy.
Area of Science:
- Biomaterials Science
- Nanotechnology
- Cancer Biology
Background:
- Cancer incidence necessitates novel therapeutic strategies.
- Gene therapy offers potential for targeted cancer treatment.
- Efficient and safe gene delivery vectors are crucial for therapeutic success.
Purpose of the Study:
- To investigate organically modified silica (ORMOSIL) nanoparticles as a non-viral gene delivery system.
- To evaluate the efficacy of ORMOSIL nanoparticles in delivering the p53 gene into breast cancer cells (MCF-7).
- To assess the anti-cancer effects of p53 gene therapy mediated by ORMOSIL nanoparticles.
Main Methods:
- Synthesis and characterization of ORMOSIL nanoparticles.
- Complexation of ORMOSIL nanoparticles with pCMV-Myc plasmid carrying the p53 gene.
- Transfection of MCF-7 breast cancer cells with ORMOSIL/p53/pCMV-Myc complex.
- Western blot analysis for p53 protein expression.
- Confocal and electron microscopy for nanoparticle localization.
- Agarose gel electrophoresis for complex stability.
- Assessment of cell survival and growth rate.
Main Results:
- ORMOSIL nanoparticles efficiently complexed with the pCMV-Myc vector carrying the p53 gene.
- Significant expression of p53 protein was observed in transfected MCF-7 cells.
- ORMOSIL nanoparticles were localized in the cytoplasm and nucleus of cancer cells.
- Transfection with ORMOSIL/p53/pCMV-Myc significantly reduced breast cancer cell survival and growth rate.
- ORMOSIL nanoparticles demonstrated effective non-viral gene delivery capabilities.
Conclusions:
- ORMOSIL nanoparticles serve as an efficient non-viral platform for gene delivery.
- p53 gene therapy mediated by ORMOSIL nanoparticles exhibits significant anti-cancer effects in vitro.
- This approach holds promise for targeted cancer gene therapy and therapeutic manipulation of cancer cells.
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