Study On Positive Charge Modified Label-free Oligonucletide Fluorescence Probe And Its Application On Anti-tumor By Traditional Chinese Medicine

Nucleic acid is very important for life, which keeps, repilicates and transmitsgenetic informations through the whole life. Nucleic acid plays a main role in growth,propagation, variation and so on. Researches on the nature and structure of nucleicacid, and interactions between nucleic acid and any other molecules are significant tounveil the secreat of life. The purpor of this study is to build a new method to detectand analyze biological macromolecules by testing comformation of nucleic acidwhich would change after interacting with some probes.Fluorescent molecular probes can convert the molecular recognition informationto the fluorescence signal which is high-sensitive to be detected. Additionally, thefluorescence signal has so many advantages such as high sensitivity, switch operation,visual submicron spatial resolution, sub millisecond time resolution, detection in situ(fluorescence imaging technology), using optical fiber to implement remote detection.Ratiometric fluorescence signal is better than fluorescent intensity response signal,which can improve the detection sensitivity and has the automatic calibration andquantitative detection functions. In this study, we synthetised a kind of pyrenefluorescent probe, and investigated its interactions with large biological molecules bynon-labeled form. And then, the pyrene fluorescent probe can be applied in biologicaldetection of living matters such as the nucleic acid and explored anti-cancermechanism.We have carried out the following work:1. We synthesized a kind of small molecule pyrene fluorescent probe, and gainedmore relevant data such as UV-visible spectroscopy, fluorescence spectroscopy,3Dspectroscopy, nuclear magnetic spectrum.2. We set up a polyanionic polymer model to simulate properties of nucleic acids.Effect of probe fluorescence detection was judged by investigating interactionsbetween different concentrations polyanionic molecule and pyrene fluorescence probe.3. We analysed the fluorescence spectrum of some nucleic acids interacted withthe pyrene fluorescent probe. We detected the effect of the concentration, the length,and the conformation of oligonucleotide on inducing PQAto form spectral peak.4. We built a kind of biological sensor to detect potassium ion. Because morethan five base oligonucleotide chain can induce pyrene fluorescent probe gathering togenerate excime, we established a kind of oligonucleotide chain that can formG-quadruplex with potassium ion. The G-quadruplex can induce pyrene fluorescentprobe gathering to generate excimer which showed potassium ion being.5. We successfully built a high-throughput cell screening model to screen thedrug activity of the Medcinal Evodia Fruit. Three single components were extractedfrom evodia rutaecarpa by screening anti-cancer activity tests. And we detected theiranti-tumor abilities.6. We analysised the anti-tumor mechanism of chinese medicines evodiamine.We tested how the Medcinal Evodia Fruit interact with human telomerase byUV-visible spectroscopy, fluorescence spectroscopy, and circular dichroism. Pyrenefluorescent probe was used in fluorescence spectroscopy detection.In this study, we built several bio-detectors which can be taken into nucleicacid identification, potassium ion detecion, and telomere research. This pyrenefluorescent probe represented many advantages such as fast, non-labeled, andlow-cost.