The Research Of Molecular Mechanism Of PDE7A In Hepatocellular Carcinoma

According to the data presently offered by the American Society of Clinical Oncology (ASCO), It’s no doubt that tumors are becoming into a global problem for our humans’health, not only in the developed countries but also in the developing countries. Meanwhile among all these kinds of tumors, the liver cancer still remains top 10, no matter in the patients of male or female. As we all know, China is a great country, where the HBV infected population is quite a problem, which can probably lead into liver cancer. Until 2015, according to the data offered by the ASCO, the morbidity of liver cancer in males in China is on the top 3, and even in females, the situation is still a serious problem. Liver cancer, nowadays, is still very hard to diagnose until it’s too bad. And there are still kinds of reasons, that even diagnosed in very early stage, we still cannot give the surgery to the patients although he or she disserve it. So it’s getting much more imperative to find more ways to rescue more patients, such as molecular targeting treatment. Phosphodiesterase including 11 isoforms can cause the hydrolysis of cAMP, and as all know the hydrolysis of cAMP can influence the regulation of the cAMP/PKA as well as the cGMP/PKC, and finally the hormones and other reactions can be regulated. Researches have already shown that PDE7A exists in kinds of cells including liver cells, and it is specific for cAMP-hydrolysis. Of cause the function of this gene can possibly change the way that is usually regulated by cAMP. All this means the study of PDE7 A may finally explain the details of the functions that PDE7A have in the hepatocellular carcinoma and definitely will contribute to the treatment of liver cancer.Part ⅠPDE7A can up regulate the proliferation of hepatocellular carcinoma cell and as well as the migrationAims:PDE7A, mostly exists in the cells of liver and brain, is a member in the families of the PDEs. Just the same like other PDEs, PDE7A can specifically hydrolyze the cAMP, so that, all those signaling pathway that controlled by the cAMP can be regulated. Abnormal cell cycle, unlimited proliferation, and outstanding migration attack ability is the common characteristics of all kinds of cancers. As it has already been studied, PDE7A has the capacity to be functioning like a real tumor gene. So in this part of articles, we try to find out what is the real influence about the cell cycle and migration in the cancer cells can be caused by PDE7A.Methods:About 90 cases of liver cancer from the first affiliated hospital of Zhejiang University have been chosen, and we collect both the tumor tissue and the normal tissue. We use the immunohistochemistry method to analysis the expression of the PDE7A, and also we compared the expression of PDE7A with the survival time of the patients. Then we analysed the expression of PDE7A in kinds of HCC cell lines to decide which kind of cells to use. Using the siRNA technology, we have made the low-expression model and the normal-expression model. Using the models, we analysed the expression of the cell cycle proteins, and the proliferation of the cells. The migration has also been tested. And the EdU and CCK8 technologies are also used to prove the effection of the expression of PDE7A.Results:The expression of PDE7A in tumor tissues is significantly high, and the low expression of PDE7A can lead the patients to a better survival time. In kinds of HCC cell lines, we found that the expression of PDE7A is higher in the HCC-LM3 cells. When we analysed the expression of cell cycle proteins wo found that in the low-expression model, the expression of ClyclinA, ClyclinD, ClyclinE and CDK2, CDK4, CDK4, are all down-regulated. And the proliferation is locked in the G2 phase. The EdU and CCK8 also show that the inhibition of PDE7A can down-regulate the cell proliferation.Conclusions:The PDE7A can be high expressed in the liver cancer, and with low expression of PDE7A the patients will have a better suvive. The low expression of PDE7A can down-regulate the expression of kinds of cell cycle proteins, and the proliferation can be locked in the G2 phase. And of course the proliferation can be inhibited.PartⅡPDE7A can intensify the Warburg effect while down-regulating the apoptosis in vivo and vitroAims:To make sure what effect can make when the expression of PDE7A is inhibited on the apoptosis, and the related proteins. To make sure what effect can make when the expression of PDE7A is inhibited on the Warburg effects. To study the exectly function of the expression of PDE7A in vivo.Methods:The FCM was used to analysis the apoptosis in HCC-LM3 cell to compare the inhibited group with the NC group. Use Western blotting to analysis the expression of proteins related to the cell apoptosis. Measure the lactic acid levels, to analysis the Warburg effect. We also use the mice model to analysis the growth of the tumor when the expression of PDE7A is inhibited, and the apoptosis, and also the changes of the Warburg effects.Results:In HCC-LM3 cells, when the PDE7A was inhibited, the apoptosis of cells is becoming larger, and as the results of the western blotting, the expression of apoptosis protein was also upregulated. Also, the lactic acid levels show the weakness of Warburg effect. At last the in vivo test shows us the inhibition of PDE7A can slow down the growth of tumor, up-regulated the apoptosis, and influence the Warburg effects.Conclusions:In these experiments, we can improve that there must be some connection between PDE7A and the apoptosis, the migration, the Warburg effect. And also shows that, PDE7A may sometimes functioning like some kind of tumor genes. The function of PDE7A can also realize in vivo and in vitro.