The Research Of Tumor Specific Activated Super Enhancer PLAU-SE

The super enhancer(SE)is a large cluster with powerful activity enhancers.It usually contains several component enhancer regions,which are larger and more powerful than ordinary enhancers and can drive important genes such as cell identity genes.SEs evolutionarily tend to exist in the vicinity of important genes related to development,usually within 20 kbp,and CTCF regions form a circular structure to limit their enhanced function on specific target genes,starting during the development of diseases such as development and tumors To the important role.Through bioinformatics methods,we selected high-level histone modifications of H3K27 ac and H3K4me1 as the primary markers in a variety of tumor cell lines,and assisted with CTCF cyclization prediction to identify multiple potential tumor-specific super enhancers..Subsequently,we knocked out the super enhancer fragment upstream of the PLAU gene by CRISPR/CAS9 gene editing technology and immediately found that knocking out this fragment specifically reduced the expression of PLAU(u-pa)and caused a series of physiological changes in He La cells.,Including lower proliferation rate,decreased migration and invasiveness,increased affinity for extracellular matrix,and increased sensitivity to cisplatin.Through the CRISPR-CAS9 technology and the fluorescence reporting system,we further defined the position of the super enhancer and the activity intensity of each component.After obtaining the cell line with the highest transcriptional activity knocked out,subsequent tumors in nude mice and Tumor staining showed that the absence of this region will reduce the degree of malignancy of Hela cells in vivo and increase the sensitivity of tumor cells to cisplatin,suggesting that this area is a potential anti-tumor targeting area.Finally,we detected the activity of the super enhancer in hypoxia and cisplatin treatment by means of chromatin immunoprecipitation(CHIP)and found that this super enhancement was induced by hypoxia(5%O2,36h).The activity of the daughter was increased,and the activity of this region was decreased by cisplatin treatment(4 ?g/ml,48 h),suggesting that the hypoxia condition may partially increase the malignant degree of the tumor cells by activating the super enhancer,and cisplatin may partially reduce the super enhancer.