The Role Of Oncogene Wip1in UVC-irradiated Glioma Cells

PartⅠThe role of p53and Wip1genes in the cell injury induced byUVC in glioma cellsObjectives The purpose of this study is to determine the survival inhibit effects ofultraviolet C（UVC）to U251(mutant type p53) and U87MG (wild type p53). Anotherpurpose is to qualify the variation of the protein expression of p53, bcl2and Wip1, and tostudy the interaction between these proteins and cell proliferation for the propermechanism.Methods Different doses of of UVC，various0,1,5,25,50and100J per m2, wereemployed to treat U87MG and U251cells. Then, the variability of the survival capabilitiesof the cells was modulated by CCK-8assay. Then, glioma cells were subjected to a dose of50J per m2of ultraviolet C irradiation. Westernblot assay was used to detect the expressionof p53and bcl2. All of the data was analysised by using SPSS10.0software.Results Ultraviolet C could harm the ability of glioma cell proliferation as well asinvasion. When the doses of ultraviolet C irradiation were between0to5J per m2, the cellproliferation of U251and U87MG had no difference; while between25and100J per m2,U251glioma cells were injuried more seriously by UVC than U87MG cells. Seventy-twohours later after being treated by a dose of50J/m2UVC, the proliferation of U87MG cellswere dropt to an extra lever: the proliferation rate of U87MG at the72hours time pointafter irradiation was（80.02±5.46）%, and the one of U251was（36.13±1.13）%. A dose of50J/m2UVC could regulate the expressions of p53and bcl2. In both of the two glioma celllines, the protein levels of p53were up-regulated within24hours, in parallel, bcl2levelsreveal the same trend. The Wip1protein levels of U251were increased in24hours, whilethe levels of U87MG were decreased at the first8hours, and had a sudden increase at24 hours after being irradiated by UVC.Conclusions The UVC-induced cell injury was more likely in U251, while U87MGcould resist it. This phenomenon may due to the function of p53protein. At the same time，Wip1may play a role in the UVC-induced glioma cell injury. Part ⅡWip1inhibitor contributes to the inhibition of glioma malignantinduced by UVC in vitroObjectives One of the purposes of this study is to determine the survival inhibit effectsof CCT007093, a Wip1inhibitor, to U251(mutant type p53) and U87MG (wild type p53).Another purpose is to qualify the variation of the expression of p53, Wip1, p38, p-p38,Wip1mRNA after glioma cell being subjected to UVC and CCT007093, and to study theinteraction between these proteins and cell proliferation, migration and invasion as well asthe proper mechanism.Methods U87MG and U251cells were treated by doses various0,5,25,50,100,200μM of CCT007093. Then, the cell survival capabilities were modulated by CCK-8assay.The cells which had been irradiated by50J/m2were then exposed to50μM CCT007093.Westernblot assay was used to detect the expression of p53, Wip1, p38and p-p38. Theexpression levels of Wip1mRNA were detected by real-time quality PCR. A Transwellassay and a Scracth migration assay were carried to study cell invasion and migration. Alldata was analysised by using SPSS10.0software.Results CCT007093could down regulate glioma cell proliferation, migration as well asinvasion, especially, in U87MG cells. When the cells were treated by a dose of50J/m2ultraviolet C irradiation and50μM CCT007093, the expression levels of p53in U251andU87MG were induced in a up-regulation within8h, but a down trend after that. p38andp-p38expressions were down-regulated in a time-dependent manner within24hours. Theexpression of Wip1protein in U251was induced constantly, but an opposite direction ofWip1mRNA. Respectively, the one was reduced in U87MG, and mRNA was induced,though within8hours only. When treated by UVC or CCT007093, U87MG had a lowerWip1mRNA expression at the24h after being treated than the8h. But when both of them, the expression of Wip1mRNA was up-regulated constantly within24hours in U87MG, to adose of (383.11±39.63)%of the expression of the control. The scratch migration assay andTranswell invasion assay revealed that Wip1inhibitor could contribute to the inhibition ofglioma cell migration and invasion induced by UVC in vitro.Conclusions CCT007093could down-regulate UVC-induced Wip1protein expression inthe wild type p53glioma cell, suppress cell migration, invasion as well as proliferation.