Expression And Role Of ZEB1 In The Human Bladder Cells And Mouse Embryonic Stem Cell Differentiation

?Purpose?ZEB1 is EMT(epithelial-interstitial transformation)important nuclear transcription factor,the role of EMT in tumor metastasis and embryonic development is more and more attention.Incidence of bladder cancer ranks first in the urinary tract cancer,and bladder cancer mortality showed a trend of increased year by year in recent years,but its mechanism is not clear.And process embryonic of stem cells differentiation has two ending,a directional differentiation into various cells in normal differentiation program,another differentiation program errors and cause cancer.Tumor cells and embryonic stem cells have certain similarity,development and embryonic stem cell differentiation and tumorigenesis mechanism also has the similarity.LIN28 A for RNA binding-protein and plays an important role in bladder cancer and embryonic stem cells differentiation,and the early stage of the experimental results show that LIN28 A plays an important role in bladder cancer cell invasion and migration,proliferation.This study selected ZEB1 as the research object,and to investigate its role in the bladder cancer as well as the relationship with LIN28 in embryonic stem cells.?Method?This research take the DNA electrophoresis,q RT-PCR and immunofluorescence technique to detect expression and location of ZEB1 in human bladder cancer cell line(UM-UC3,5637),the mouse embryonic stem cells(m ES)mouse embryoid body(m EB)GC1cells and mouse embryonal carcinoma cells(P19).Expression of ETM related proteins(E-cadhern?Vimentin)and LIN28 were conformed by q RT-PCR and Western Blot after ZEB1 knockdown.The effect of ZEB1 on tumor cell invasion was observed by Transwell invasion assays in the human bladder cancer cells and mouse embryonal carcinoma cells.?Results?ZEB1 in human bladder cell line(UM-UC3,5637),m ES,m EB,GC1,P19 are expressed and expressed in malignant degree higher UM-UC3 quantity higher than that of low degree of malignant cell line 5637,And differentiation of mouse embryonic stem cells in the normal way,ZEB1 reduced gradually,when abnormal way of embryonal carcinoma,P19 cells expressing quantity is GC1 eight times.ZEB1 proteins were localized in the nucleus.After ZEB1 was knocked down in the bladder as the mouse embryonal carcinoma cell invasive rate reduced.?Conclusions?ZEB1 expressed in bladder cancer cell line(UM-UC3 and 5637)and ZEB1 promotecancer cells,E-cadherin expression increased,Vimentin expression.UM-UC3 cell line : the ratio of groups of cells invade in the group of ZEB1 knock down compared with the wild group WT and the negative control group NC was reduced by 72% and 68%(* P<0.05).5637 cells line : the ratio of groups of cells invade in the group of ZEB1 knock down compared with the wild group WT and the negative control group NC was reduced by 50% and 40%(* P<0.05).After ZEB1 was knocked down in m ES?m EB?GC1?P19 cells line,E-cadhern expression increased,Vimentin expression reduced.LIN28 Am RNA express fell by 44%,51%,37%,51%(* P < 0.05)and LIN28 A protein expression quantity decreased.The ratio of groups of cells invade in the group of ZEB1 knock down compared with the wild group WT and the negative control group NC was reduced by 49.8% and 50%(* P<0.05)in mouse embryonal carcinoma cells(P19)?Conclusions?ZEB1 expressed in bladder cancer cell line(UM-UC3 and 5637)and ZEB1 promote the invasion of bladder cancer cells by EMT.ZEB1 expressed in normal or abnormal differentiation of embryonic stem cells,Expressed in normal directional differentiation route gradually reduce,in the abnormal differentiation approach for embryonal carcinoma P19,ZEB1 levels higher than GC1 expression.ZEB1 through regulating LIN28 and EMT process involved in differentiation of embryonic stem cells.