Ansari Anees A, Hasan T N, Syed N A, Labis J P, Alshatwi Ali A
King Abdullah Institute for Nanotechnology, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia.
Molecular Cancer Biology Research Lab (MCBRL), Department of Food Science and Nutrition, King Saud University, Riyadh 11451, Saudi Arabia.
Saudi J Biol Sci. 2017 Sep;24(6):1392-1403. doi: 10.1016/j.sjbs.2016.08.012. Epub 2016 Sep 8.
Monodispersed luminescent functionalized core-shell nanospheres (LFCSNs) were successfully synthesized and investigated for their cyto-toxic effect on human liver hepatocellular carcinoma cell line (HepG2 cells) by adopting MTT, DNA Ladder, TUNEL assay and qPCR based gene expressions through mRNA quantifications. The TUNEL and DNA ladder assays suggested an insignificant apoptosis in HepG2 cells due to the LFCSNs treatment. Further, the qPCR results also show that the mRNA expressions of cell cycle checkpoint gene p53 and apoptosis related gene () was up-regulated, while the antiapoptotic gene and apoptosis related genes and (apoptosis effecter gene) were down-regulated in the LFCSNs treated cells. The nanospheres that were loaded into the cells confirm their intracellular uptake by light and fluorescent spectro-photometry and microscopy imaging analysis. The loaded nanospheres demonstrate an absolute resistance to photo-bleaching, which were applied for dynamic imaging to real-time tracking - cell migratory activity for continuous 24 and 48 h durations using a time-lapsed fluorescent microscope. These properties of LFCSNs could therefore promote applications in the area of fluorescent protein biolabeling and drug-delivery.
通过MTT法、DNA Ladder法、TUNEL检测法以及基于mRNA定量的qPCR基因表达分析，成功合成了单分散发光功能化核壳纳米球（LFCSNs），并研究了其对人肝癌细胞系（HepG2细胞）的细胞毒性作用。TUNEL检测法和DNA Ladder法表明，LFCSNs处理后HepG2细胞中的凋亡不明显。此外，qPCR结果还显示，在经LFCSNs处理的细胞中，细胞周期检查点基因p53和凋亡相关基因（）的mRNA表达上调，而抗凋亡基因以及凋亡相关基因和（凋亡效应基因）的表达下调。通过光、荧光分光光度法和显微镜成像分析证实，进入细胞的纳米球被细胞摄取。负载的纳米球表现出绝对的抗光漂白性，使用延时荧光显微镜对其进行动态成像，以实时跟踪细胞在24小时和48小时内的迁移活性。因此，LFCSNs的这些特性可促进其在荧光蛋白生物标记和药物递送领域的应用。
