| Gastric cancer is one of the most common malignant tumors threating to human health. A few cells escaped from the original focus and disseminated into other organs is a common reason leading to the death of gastric cancer patients. Therefore, the detection and imaging of gastric cancer cells are of great clinical significance for effectively preventing and treating gastric cancer. We have carried out some works for gastric cancer cells recognization and detection by double fluorescence labeling for confocal laser scanning microscopy imaging (CLSM), which employed luminescent and photostable RuBpy doped silica nanoparticles (RFSiNPs) and FITC doped silica nanoparticles (FFSiNPs) as fluorescence labels. The three works are as follows.1. In vitro imaging of MGC-803 cells by bioconjugated FSiNPs mediated double immunofluorescence staining.Based on immunolabelling technique, we have used anti-CEA antibody conjugated FFSiNPs and anti-CK19 antibody conjugated RFSiNPs to diagnose gastric cancer related antigen CEA and CK19 simultaneously. We developed a fluorescent silica nanoparticles (FSiNPs) mediated double immunofluorescence staining technique for MGC-803 gastric cancer cells imaging by confocal laser scanning microscopy. The results show that the method has superior selectivity and can greatly improve the accuracy of tumor cell detection. The different cells can be accurately distinguished via their specific antigen expressions. Comparing with the conventional double immunofluorescence staining using green-emitting and red-emitting dyes, the photostability of this proposed method for CLSM imaging has been greatly improved.2. Imaging of MGC-803 cells from peripheral blood and in vivo by bioconjugated FSiNPs mediated immunofluorescence staining.In order to investigate the feasibility of diagnosing gastric metastasis in peripheral blood, bioconjugated FSiNPs mediated immunofluorescence staining and flow cytometry are used to detect MGC-803 gastric cancer cells in peripheral blood. On the basis of gastric cancer nude mice model, the ex-vivo imaging of primary MGC-803 gastric cancer cells samples from the tumor tissues of mice bearing the MGC gastric cancer tumor xenografts by this method has also been explored. Furthermore, the anti-CK19 antibody conjugated RFSiNPs were injected into the mice for gastric cells imging in vivo. Results show that immunofluorescence double label methods has a well adaptability to complex biological system and can specifically rcognize a few MGC-803 cells in peripheral blood, which is expected to be used in early tumor diagnosis and cancer metastasis research. By tumor imaging in vivo, we found that RFSiNPs can be gathered in the tumor region and thus formed good imaging results, which showed the potential of FSiNPs in vivo imaging.3,Study on cellular uptake of fluorescent silica nanoparticles by mice kidney cells.The excretion of silica nanoparticles in vivo was selected as the research object because of the potential application of silica nanoparticles in vivo imaging. By using fluorescent signal of RFSiNPs, the cellular uptake of fluorescent silica nanoparticles by MC cells and NRK cells has been investigated. Results showed that both of the two kinds of cells have the capability of endocytosis of fluorescent silica nanoparticles, and the endocytosis amount increased obviously with increase of nanoparticle concentration. In addition, the serum in cell culture medium can also affect the cellular uptake. Compared with the HUVEC cell, NRK cells have a stronger capability of endocytosis, which may be owing to the physiological functions of mice kidney cells. This work has provided some useful information for the research on the cellular uptake of silica nanoparticles with mice kidney cells.
|
PDF Full Text Request