Rejeeth Chandrababu, Kannan Soundarapandian, Muthuchelian Krishnasamy
Proteomics and Molecular Cell Physiology Lab, Department of Zoology, School of Life Sciences, Bharathiar University, Coimbatore, 641 046, TN India.
Department of Energy, School of Biological Sciences, Madurai Kamaraj University, Madurai, TN India.
Cancer Nanotechnol. 2012;3(1-6):55-63. doi: 10.1007/s12645-012-0030-3. Epub 2012 Jul 29.
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.
本文报道了近年来有机改性二氧化硅(ORMOSIL)纳米颗粒作为一种高效的体外基因传递系统的应用。基于这一主要目标,本研究探讨了在细胞水平上降低癌症发病率的可能方法。在此背景下,合成了ORMOSIL纳米颗粒,并将其与携带p53基因的pCMV-Myc(3.8 kb)质粒载体构建体一起孵育,然后转染到乳腺癌细胞系MCF-7细胞中。蛋白质免疫印迹分析表明,转染后p53蛋白在乳腺癌细胞中显著表达。共聚焦显微镜和电子显微镜研究进一步证实,纳米颗粒在转染了p53基因的癌细胞的细胞质和细胞核中积累。有趣的是,琼脂糖凝胶电泳研究表明,纳米颗粒能与pCMV-Myc载体有效复合。通过评估细胞存活率和生长速率证明了p53的抗癌特性,结果显示在癌细胞中呈正线性相关。然而,与未转染细胞的生长速率相比,ORMOSIL/p53/pCMV-Myc转染的乳腺癌细胞的生长速率显著降低。使用ORMOSIL纳米颗粒作为非病毒基因传递平台的这种方法的结果对于用作癌细胞治疗操作的有效转染剂和体内靶向癌症基因治疗具有广阔的前景。