Effect Of TET2 Regulation Of MAP4K1 Expression On Migration And Apoptosis Of T24 Cells

Object:To investigate TET2 regulation expression of MAP4K1and the migration and apoptosis of T24 cells in bladder urothelial cell carcinoma T24,and to provide experiment basis for the diagnosis and treatment of bladder cancer.Materials and methods:(1)Materials:Bladder urothelial cancer cell line(T24)was selected as the research object.(2)Methods:TET2overexpression and suppression expression vectors and corresponding negative control vectors into were transfeted into bladder urothelial cancer cell line(T24),the expression levels of TET2 and MAP4K1proteins were detected by Western Blot,The migration ability of T24cells was detected by cell scratch test,The apoptosis of T24 cells was detected by flow cytomety.The experiment groups were divided into the overexpression group(T24 cells+overexpression TET2 plasmid group)and the negative control group(T24 cells+empty plasmid group)and the inhibition group(T24 cells+si-TET2 group)and the negative control group(T24 cells+si-TET2-NC group)and the blank control group(T24 cells group).The TET2 overexpression plasmid,empty plasmid,si-TET2fragment and TET2-NC fragment synthesized in vitro were transfected into T24 cells of overexpression group and negative control group,inhibition group and negative control group respectively by lipofectaminet~TMM 2000.After 6 hours of incubation,the transfection efficiency of the cells reached more than 80%by comparing the fluorescence microscope field of view with the light microscope field of view,and subsequent experiments can be carried out.The protein expression of TET2 and MAP4K1 was detected by Western Blot,the cell migration ability of each group was detected by cell scratch test,and the apoptosis rate of each group was detected by flow cytology.(2)Statistical methods:the data were processed and analyzed by SPSS23.0 statistical software and EXCEL software.the measurement data were expressed as mean standard deviation(?±s).the comparison of measurement mean between the two groups was conducted by t test,the data have statistical significance.with p<0.05.Results:(1)Cell transfection:TET2 plasmid and TET2-siRNA were transfected into T24 cells successfully;(2)WB experiment:Compared with TET2 overexpression control group,in TET2 overexpression group,the expression level of TET2 protein was significantly increased,while the expression level of MAP4K1 protein was significantly decreased;Compared with the control group of TET2 inhibition group,the TET2 expression content in TET2inhibition group was obviously decreased,while the protein expression content of MAP4K1 was obviously increased;Compared with TET2 overexpression control group and TET2 inhibition control group,the blank group showed no significant difference;(3)Cell scratch test:Compared with the control group of TET2 inhibition group,the TET2 expression content in TET2 inhibition group was obviously decreased,while the protein expression content of MAP4K1 was obviously increased;Compared with the control group of TET2 inhibition group,the healing rate of TET2 inhibition group was significantly higher;Compared with TET2 overexpression control group and TET2 inhibition control group,the blank group showed no significant difference;(4)Apoptosis experiment:Compared with TET2 overexpression control group,the apoptosis rate of TET2 overexpression group was lower;Compared with the control group of TET2 inhibition group,the apoptosis rate of TET2 inhibition group was significantly higher;Compared with TET2 overexpression control group and TET2inhibition control group,the blank group showed no significant difference.Conclusion:TET2 down-regulate the expression of MAP4K1 and inhibit the migration of T24 cells.