The YAP1 Proteins Confers Cancer Stem Cell-Related Traits On Prostate Cancer Cells

Objective: To detection the expression of Yes-associated protein(YAP)in the human prostate cancer stem cells.Using the Screening of cancer stem cells and non-stem cells that transfected by YAP1,observe their self-renewal and proliferation in vivo and in vitro phase changes.Then investigate the role of YAP1 in tumor progression and its’ effects in regulating cells’ polarity.Methods: Using of prostate cancer stem cell marker CD133 antibodies marked tumor cells.In the paper,we sort the prostate cancer cell line LNCa P cells and C42 cells using magnetic separator(MACS),filter out cancer stem cells(CD133 positive)and non-tumor stem cells(negative CD133).Next we using Western blot,to detecte the expression of YAP1 in tumor(non-stem cells,cancer stem cells,General).CD133 bead filter cells are conducted in further experiments,we stably transfected with the plasmid,in cancer stem cells knocking out YAP1,in non-tumor stem cells overexpression of YAP1,then we conduct colony forming experiments,to observe the changes of gene-modified cells’ self-renewal and proliferation capacity.Then we conducted the experiments in vivo,injecting the cells subcutaneously in nude mice,measure the diameter of the tumor every 7 days,then harvesting the tumor.we treat the sample in immunohistochemistry,to observe and validate the expression of CD133,YAP1,Ki67,SOX2,OCT4.Results: 1.The expression of YAP1 is higher in CD133(+)cells than in CD133(-)cells.2.The CD133(-)cells,that is overexpression of YAP1,have a higher colony formation rates than CD133(+)group,that is knocked out YAP1.3.In vivo,the CD133(-)cells’ tumor grow fastest than others in nude mice subcutaneous.4.In the immunohistochemistry,the expression of the CD133,YAP1,Ki67,Sox2,Oct4 are higer in the CD133(-)cells’ tumor,that is knocked out YAP1.Conclusions: Activation of the YAP1 will increase the number of non-stem cells transfering into cancer stem cells,but the mechanism is not yet understood,it will serve as an important therapeutic target in the clinical.