The Synthesis And Diagnosis Effection Of Fluorescent Probes Which Targeting COX-2

There are 700 thousand new cases of Primary liver cancer in the world each year and it has become a serious impact on the life and health of people.Primary liver cancer including liver cells cancer, cholangiocarcinoma, hepatoblastoma, and angiosarcoma and in primary liver cancer, hepatocellular carcinoma(HCC) accounted for the proportionmore than 80%.And the incidence of liver cancer in Asia, especially in China, was significantly higher than that in the world. There are reports that the entire world of patients with hepatocellular carcinoma more then 50% concentrated in china.The major risk factors for the hepatocellular carcinoma are hepatitis virus infection(in our country mainly of hepatitis B virus infection), a large number of drinking, long-term intake of aflatoxin contamination of food, obesity, smoking, fatty liver, diabetes and so on.In different countries and regions, the main risk factors of primary hepatocellular carcinoma are often not the same, and that make it difficult for the clinical diagnosis, treatment and prognosis of hepatocellular carcinoma According to the literature, at present, the mortality of hepatocellular carcinoma in our country has accounted for 18.8% of all cancer in our country. The standard treatment of liver cancer is still the comprehensive treatment based on surgery.Surgical resection is considered to the first choice of treatment for patients with hepatocellular carcinoma, 5 years survival rate can reach 60%-70%, some patients can be cure.With the progress of preoperative imaging and laboratory examination,more preoperative assessments such as child Pugh liver function grade, indocyanine green clearance experiment(icg15) and hepatic venous pressure gradient detection are tested for the patient.With the innovation and progress of medical technology and medical devices, such as ultrasonic scalpel, argon knife, regional hepatic blood flow blocking technology development, improvement of the anatomical structure of the liver and clinicians operation level, the perioperative period mortality of liver cancer patients decreased significantly, several major medical centerscan control the mortality rate to 1%- 5%.However, the long-term prognosis of patients with hepatocellular carcinoma is still not satisfied. Cause there are a lot of factors of poor prognosis of hepatocellular carcinoma: no clear clinical symptoms In the early stage of the disease. When there are clinical symptoms and go to a doctor the diseasehave been developed, not sensitive to chemotherapy and radiation therapies, The high recurrence rate after surgical resection and tend to occur earlier transfer and so on.In these reasons, the most common cause of recurrence are Primary tumor and metastatic lesions residual during the operation.The postoperative recurrence of hepatocellular carcinoma has a serious impact on the treatment outcome and prognosis of the patients.However, the judgment of these two aspects often need a Certain tumor size and a clear metastatic.And this kind of judgment is often based on the experience of clinicians.How to find a method that can help the clinicians to find out the extent of the tumor and determinethe surgical resection specimen more clearly, is the problem that need to be solved.With the developments of the fluorescent phenomenon and various specific fluorescent dyes, the application of the fluorescence detection technology in the field of life science has become more and more widely.To identifysome specific indicatorswithin certain cells or organelles and trans these micro signals by some means into the macroscopic signals that can be tested by instruments called biologicalidentification.Biomarkers is a kind of special type of biological products which can be used in the biological identification of a class of cells, tissue growth, differentiation, proliferation, degenerationand apoptosis.In the field of biological research, biomarkers have been widely used in cytology, oncology, biological metabolism and other related fields.Because of its physiological properties and the way of microstructure expressionof, biomarkers have high accuracy and sensitivity characteristics, biomarkers can be changed in the early perior of biological behavior that at the time has not showobvious macroscopic changes.By some special equipment and technology, the microscopic changes of biomarkers can be observed, and can achieve the identification of biological molecules, the diagnosis of specific diseases, as well as for the purpose of biological treatment of some target phase point.The combination of biological recognition technology and fluorescence color rendering technology constitutes fluorescent probe technique.fluorescence probecan emit fluorescence in the ultraviolet-visible-near infrared region,and its fluorescence properties(excitation and emission wavelength, intensity, lifetime, polarization, etc.) can be changed with the nature of the environment, such as polarity, refractive index, viscosity and other changes.At present, in biological research, fluorescent probes can combinate with nucleic acid(DNA or RNA), protein or other macromolecular structure and emit a specific fluorescence.COX-2 is an inducible subunit of the epoxy synthase, which is rarely expressed or not expressed in normal tissues. It is highly expressed by the external stimuli such as inflammation, tumor and so on.At present, that has found there is a high expression of COX-2 in gastric cancer, liver cancer, breast cancer, colon cancer and other tumors, and a number of studies had proved that COX-2 play an important role in the biological behavior of the tumor occurrence, development and metastasis.So,COX-2 can be used as a tumor marker for tumor detection and identification.In this study, the fluorescent probes were used to label the high expression of COX-2 in liver cancer tissue, so that it can emit fluorescence in the range of visible light wavelength.It provides a new idea for the study of the biological behavior of hepatocellular carcinoma and the experimental diagnosis of primary tumor and the range of metastasis.Objective: To synthesize the fluorescent probes which can be targeted to COX-2 and apply them tothe cell, tissue and animal model of liver cancer, and to study its clinical application value.Methods: 1. Synthesize the fluorescent probes withfluorescence groups and COX-2 distinguish groups. 2. Determinat the light stability and COX-2 recognition specificity of the fluorescent Probe. 3. Useimmunohistochemical method to find out the expression change of COX-2 in humanhepatocellular carcinoma cells SMMC7721, BEL7402, and normal liver cells LO2. 4. Fluorescent probe X was used to stain the cells. 5.Provethe fluorescence probe stainethe tumor cells by the target of COX-2. 6. Verify the cytotoxicity of fluorescent probes. 7. Made the nude mouse model of human hepatocellular carcinoma and do fluorescent stain. 8.The surgical resection of liver cancer tissue and normal liver tissue were stain and compared.Results: 1. Fluorescent probe X can be achieved through the combination of COX-2 protein to emit fluorescence and has good stability. 2. Human hepatocellular carcinoma cells SMMC7721, BEL7402 highly express COX-2 and normal liver cells LO2 does not express COX-2. 3. Fluorescent probes can be used to distinguish hepatocellular carcinoma cells and normal hepatocytes. 4.The Fluorescence difference of liver cancer cells and normal liver cells is achieved through the specificity of COX-2. 5. No significant cytotoxicity was observed in the cell experiments. 6. Ues fluorescence probe stain human hepatocellular carcinoma transfer to nude mice mode.7. Fluorescence probe was able to distinguish tumor tissues and normal liver tissue from surgical resection.Conclusion: The fluorescent probe X can be target to the COX-2 protein and emit fluorescence in hepatocellular carcinoma and transfer tumor in nude mice model and has importment value in clinical application.