Hspb1Expression On Tumor Cell Proliferation In The U251Cell Line And Differences In The Stem Cell

ObjectiveHeat shock protein family (HSPs) in U251cell lines and glioma stemcell differences in expression to explore to explore the expression of heatshock protein B1in the U251cell line and its stem cells, to observe theimpact of tumor cell proliferationMethods1To adopt DMEM/DF12serum-free medium to cultivate U251cells,get U251stem cells.2The application of immunofluorescence staining bydetecting CD133and nestin, and thus the identification of glioma stemcells.3The application of iTRAQ technology HSPs difference analysis on theU251cell line and its stem cells.4Choice of different concentrations ofcarmustine on U251cell line, intervention, and screening a suitableconcentration.5U251cell line, U251cell lines+carmustine, U251stem cells,stem cells of U251+carmustine is divided into four experimental groups,MTT assay was used by Western blot and RT-PCR (Real-time PCR) wasobserved by the carmustine after the intervention, tumor cell apoptosis and HSPB1protein, gene expression changes in the relationship, and used forstatistical analysis.Results12-3weeks in serum free medium, the cell ball. Two markers CD133and Nestin U251stem cells were identified, while the expression of CD133and Nestin cells more than95%.2U251stem cells and U251cells, there are four heat shock proteinsshowing differential expression, respectively HSPA5(IPI00003362.2),HSPB1(IPI00025512.2) HSP90B1(IPI00027230.3), HSPE1(IPI00220362.5), the ratios were0.423,0.523,0.522,1.484.3The carmustine inhibitory rate of U251cells, the choice of6.25μmol/L concentration of carmustine experimental conditions.5After6.25μmol/Lcarmustine intervention U251and its stem cells, HSPB1protein contentwere increased by1.4,3.3times HSPB1mRNA were increased by1.5,3.6times the intervention of the same concentration of carmustine,U251cellgrowth was inhibited, while the U251stem cells were not suppressed.ConclusionThe HSPB1stem cell chemotherapy resistance to the process involvedin U251glioma tumor cell proliferation, apoptosis plays an important role inthe regulation.