| Background:Primary liver cancer is the most common malignant tumor in the digestive system all over the world.It is ranked fifth in the cancer incidence.Its tumor related mortality was third in the malignant tumor,and its incidence is still increasing.More than 150 million new liver cancer patients worldwide each year,of which about half of our country.In treatment of liver cancer method has been developed for surgical treatment is the main therapy,radiofrequency ablation,interventional treatment,chemotherapy,radiotherapy,target gene to treatment,supplemented by a comprehensive treatment program,in a certain extent,improve the cure rate of liver cancer.But the liver cancer in our country is still the second largest tumor mortality after lung cancer.The main reason is the insidious onset of liver cancer at the time of diagnosis has been developed as the most advanced,lost the best surgical treatment opportunities.In addition,the liver resulting in highly heterogeneous conventional radiotherapy and chemotherapy is not sensitive,so easy to relapse after liver cancer.In order to improve the prognosis of patients with hepatocellular carcinoma,improve the cure rate of the disease,we need to find an efficient,stable,side effects of liver cancer drug intervention targets for clinical practice.NF-?B is a key gene transcription regulatory factor in the human body,in most cases,NF-?b and its inhibitory factor I?B in the cytoplasm in the inactive state.Only in its being such as ray,chemicals,viruses,bacteria,upstream stimulatory factor activator activation after will and I?B phase dissociation exposed binding bridge,and across the nuclear membrane into the nucleus guide cell cycle and DNA replication,transcription and other aspects,so as to control the proliferation and apoptosis of the cells.NF-?B is rarely expressed in most cells of the human body,and is in the inactive state after being combined with the inhibiting factor I?B.Existing studies have shown that NF-?B expression in tumor cells is several times higher than normal cells,and this has been verified in lung cancer,breast cancer,prostate cancer,colorectal cancer,pancreatic cancer and other different tumors.In the studies of liver cancer has also been similar conclusions,namely in the early stage of hepatocarcinogenesis induced,with proliferation of tumor progression,NF-?B expression showed a linear increase trend,reaching a peak in liver cell cancer thoroughly,and has been in a high expression.Many experiments show that artificially drug intervention to tumor cells in the NF-?B inactivation,can greatly reduce the content of tumor associated markers,and the tumor growth at a standstill,the inhibition of tumor invasion and metastasis.Toll like receptor 4 signaling pathway is one of the focuses of current research in oncology.In recent years,more and more of the view that it is not just recognition and transmission of signals within the cell-mediated,but involved in the development of inflammation,oxygen free radicals and important signaling pathways induced immune function in cancer immune escape mechanisms It has a pivotal role.MyD88 in TLR4 signaling pathway in the narrow part of the public,played a key role in the convergence of upstream and downstream signals.The chemical nature of MyD88 is a soluble protein,acts as a switch in TLR4 signaling pathway.In the TLR4 pathway,upstream of the Toll-like receptor and MyD88 Toll-like area combine to make MyD88 activation and aided by downstream signal transduction start in IL-1 receptor to promote the phosphorylation of I?B from NF-?B-I?B dissociates combination.Eventually leading to abnormal activation of NF-?B,exposing binding bridge and into the abnormal expression of proto-oncogene to start the nucleus and induce liver cancer.ST2825 is a specific inhibitor of Myd88.Experiments confirmed ST2825 by competitive occupy Myd88 Toll-like receptor-binding region thereby inhibiting signal transduction upstream and interleukin-1-related protein kinase activity,and thus silence the abnormal expression of NF-?B.ST2825 is not easy to be cleared by the body's enzymatic hydrolysis system,which has the advantage of natural MyD88 inhibitors.In the inhibition of MyD88 activity with high efficiency,stability and other characteristics.In many malignant tumors and inflammatory stimulation,immune induction it has been taken as a new drug target to study,has made an important breakthrough.But it has not been reported in the research of prevention and treatment of liver cancer.Based on the above theories,we have reasons to establish such a hypothesis:ST2825 able to intervene human liver cancer occurrence,development,invasion and metastasis,inhibit liver cancer conventional development process,improve the survival rate of patients with hepatocellular carcinoma.Our subject from two angles in the body and digress to experiment in order to discover new drugs effective intervention HCC progression targets.Objectives:1.To identify the expression of Myd88 in liver cancer and adjacent normal tissues.2.To identify the the proliferation and apoptosis of hepatocellular carcinoma cells and the growth of transplanted tumor in nude mice after application of ST2825Methods:1.Immunohistochemistry,RT-PCR was used to detect the expression of liver cancer and adjacent normal tissue samples whether MyD88 gene expression in hepatocellular carcinoma was significantly higher than in adjacent normal tissue.2.Using the RT-PCR and Western blot to investigate the role of human hepatoma cell changes MyD88 gene after application different concentrations of ST2825.3.Using the flow cytometry technique,MTT method and Western blot to assay hepatoma cells proliferation and apoptosis changes after ST2825's effect.4.Test the effect of ST2825 on the growth of transplanted human hepatocellular carcinoma cells in nude miceResult:1.The results of immunohistochemistry showed that the expression of MyD88 in liver cancer was significantly higher than that in adjacent normal tissues.RT-PCR results showed that the expression of m RNA MyD88 in liver cancer was significantly higher than that in adjacent normal tissues.2.The results of RT-PCR and blot Western showed that the expression of MyD88 gene in human hepatocellular carcinoma cells at different concentration of ST2825 was decreased in different degrees,and the higher the ST2825 concentration,the lower the MyD88 expression,all of which had statistical significance.3.The results of flow cytometry showed that the proportion of early apoptosis of liver cancer cells was significantly increased,and the proportion of GO/G1 phase cells was significantly increased after ST2825.4.The expression of apoptosis related protein caspase-3 in human hepatocellular carcinoma cells was significantly increased,and the expression of anti apoptotic protein Bcl-2 was significantly decreased by using ST2825.The expression of proliferation associated protein cyclin D1,P65 significantly decreased the expression of I?B increased significantly.5.The detection results of MTT method showed that the activity of human hepatocellular carcinoma cells in ST2825 was significantly decreased in dose-dependent manner.6.In vivo experiments showed that ST2825 could inhibit the growth of transplanted tumor in nude mice.Conclusions:1.The expression of MyD88 in liver cancer tissue was significantly higher than that in normal tissue,and the difference was statistically significant.2.The proliferation of human hepatocellular carcinoma Hep G-2 cells was inhibited and the early apoptosis was promoted by ST2825,and the difference was statistically significant.3.ST2825 can significantly inhibit the growth of human hepatocellular carcinoma in nude mice,the inhibition effect was dose dependent. |
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