The Interaction Of DEK Protein With Telomerase

PURPOSE:The DEK protein is a chromatin architectural protein, in cervical cancer the level of DEK protein is significantly higher than that of normal cervical cells. And its specific overexpression is observed in a variety of tumors, so it has drawn a great deal of attention in recent years. With a detailed study of the DEK protein, there is progress step by step, but till now its mechanism is still not very clear. We hope to reveal the relationship between the DEK protein and telomerase, which also takes a lot of attention in tumor. So the mechanism of the DEK protein in tumors could be further studied.METHODS: We selected the Hela cell line as a study sample, and used the eukaryotic expression vector pcDNA3.1to construct pcDNA3.1-His-DEK. For transfection, we chose FuGENE HD transfection reagent from Roche. The shRNA vectors are synthesized by a company, respectively, which can introduce the intracellular expression of siRNA of DEK oncogene, and they could silence DEK oncogene.Based on the determination of experimental conditions for the transfection, we used Western Blot and immunohistochemistry to detect the DEK protein levels in the treated cells, and TRAP-PCR kit was used to detect telomerase activity. Quantitative PCR method was used to detect the mRNA levels of DEK gene and hTERT gene, so we can try to determine the relationship between DEK and telomerase. Also the expression level of the p53gene and MCL-1gene was detected. We wish that we could find out the linkage of these two tumor related genes with DEK and telomerase.RESULTS: We successful detected overexpression of DEK protein in Hela cells by Western Blot and immunohistochemistry, which were transfected with DEK expression vector, and detected the reduction of DEK protein in Hela cells transfected with shRNA vector. Detection of telomerase activity failed to see the telomerase activity in Hela cells. Using qPCR, we could observe some changes in DEK expression, hTRET and p53showed decrease in cells transfected with shRNA2#plasmid. It showed that large number of MCL-1was expressed in the cells, and it decreased when DEK expression is decreased.