| Summary: | 碩士 === 長庚大學 === 生物醫學研究所 === 104 === Dengue fever is an important human viral disease that is transmitted by mosquito vectors, primarily Aedes aegypi and Aedes albopictus. The dengue virus can be divided into four serotypes (DEN1, DEN2, DEN3 and DEN4). According to the lab past results, mammalian cells usually cause cell death was they are infected with dengue virus. In contrast, mosquito cells infected with dengue virus frequently cause increased oxidative stress without the outcome of cell death. The p53 is a tumor suppressor gene, which is usually involved in inducing DNA repair and, cell cycle arrest as well as apoptosis when the cells respond tp the outside stress. The p53 level can affect the intracellular oxidative stress. In this study, we aim to investigate the role of p53 in C6/36 cells with DEN2 virus infection. First, we have identified the p53-like sequence from the C6/36 cells and named it as p53-2. Then, we obtained the other one Aedes albopictus p53 sequence from the database and called it p53-1. Based on the Bioinformatics analysis, we confirmed that the p53-2 sequence of Aedes albopictus belonges to the p53 family and had 40% similarity with p53-1. It revealted there are two p53 paralogs existing in Aedes albopictus. Next, we determined to the level of two p53 sequences in C6/36 cells with DEN2 infection, showing its expression level increased to 300%, but the p53-1 remained at the same level. Subsequently, we explored the function of p53-2, showin that knockdown of p53-2 enhanced increased accumulation of the reactive oxygen species (ROS) and cell death in DEN2-infected C6/36 cells. At last, we used the pAC5.1-A-neob-p53-HA to over-express p53-2 in C6/36 cells which have knocked down the expression of p53-2, leading to recovery of the p53-2 level to the physiological level. This will be used as a tool for future study in aossociated experiments. In summary, we have identified a new paralog of p53 in the mosquito Aedes albopictue; which is eventually upregulated in response to DEN2 infection in C6/36 cells and created a model for investigation on p53-regulated antioxidant defense in C6/36 cells when they are infected by the dengue virus.
|
|---|
