The Effect Of MAP4K1 Expression Levels On The Proliferation,Invasion And Migration Of T24 Cells In Bladder Urothelial Carcinoma

Objective:To investigate the effect of the expression of MAP4K1 on the proliferation,invasion and migration of T24 cells in bladder urothelial carcinoma,and to provide new ideas for the diagnosis and treatment of bladder cancer.Materials and Methods:(1)T24 cell,the bladder urinary epithelial cancer cell line,was used for the research.The experimental groups were as follows: blank control group(T24 cell group),negative control group(T24 cell + MAP4K1-NC group),MAP4K1 inhibition group(T24 cell + MAP4K1-homo-524),MAP4K1 overexpression group(T24 + MAP4K1 overexpression vector).After inoculation for 24 hours,the cells were transfected with MAP4K1-NC fragment,MAP4K1-homo-524 fragment and MAP4K1 overexpression vector in vitro by Lipofectamine TM 2000 in T24 cells of negative control group,inhibition group and overexpression group respectively.(1)The transfection efficiency of each group was observed under the fluorescence microscope after transfection of 6h.And the transfection efficiency of cells reached more-than 80%,and the follow-up experiments could be carried out.(2)After transfection,the OD value of cells in each group was detected by MTT assay at 24 h,48h,72 h.(3)Transwell experiments were performed after transfection of 24 h,to detect the number of transmembrane cells in each group.(4)Cells were stained after transfection of 24 h,and cultured for 24 h,48h,72 h.The wound healing rate of each group was detected by scratch test.Result:(1)The transfected T24 cells were observed under the fluorescence microscope,and when the transfection efficiency of cells in each transfected group were more than 80%,the follow-up experiments could be carried out.(2)Effect of MAP4K1 expression level on proliferation of T24 cells: the MTT results showed that: the OD values of MAP4K1 overexpression group were 0.601 ± 0.021,0.822 ± 0.029 and 1.012 ± 0.039 respectively at 24 hours,48 hours and 72 hours after transfection.The proliferation rates were 7.5%±0.4%,11.8%±0.3% and 14.1% ±0.9%respectively.There were significant difference among MAP4K1 overexpression group and blank control group and negative control group,(P< 0.05).The OD values of MAP4K1 inhibition group were 0.484 ± 0.026,0.625 ± 0.028 and 0.694 ± 0.042 respectively at 24 hours,48 hours and 72 hours after transfection;The inhibition rates were: 10.5%±0.6%,14.3%±0.8%,16.8±1.0%.There were significant difference among MAP4K1 inhibition group and blank control group and negative control group,(P< 0.05).There was no significant difference between blank control group and negative control group(P> 0.05).(3)Effect of MAP4K1 expression level on invasiveness of T24 cells: The results of Transwell invasion experiment showed that,after transfection for 24 hours,the number of transmembrane cells in MAP4K1 overexpression group was 45±2.5.There were significant difference among MAP4K1 overexpression group and blank control group and negative control group,(P< 0.05).And the number of transmembrane cells in MAP4K1 repressor group was 28± 2.0 and 35±1.8 in blank control group and 38±1.6 in negative control group respectively.There were significant difference among MAP4K1 inhibition group and blank control group and negative control group,(P< 0.05).There was no significant difference between blank control group and negative control group(P> 0.05).(4)Effect of MAP4K1 expression level on T24 cell migration: Scratched 24 hours later,the scratch healing rates were 8.39% ± 0.55% in blank control group and 7.90% ±0.89 % in negative control group and 14.73% ± 0.84% in MAP4K1 overexpression group and 5.16% ± 0.75% in MAP4K1 inhibition group respectively.There were significant difference among MAP4K1 overexpression group and blank control group and negative control group,(P< 0.05)and also among MAP4K1 inhibition group and blank control group and negative control group,(P< 0.05).Scratched 48 hours later,the scratch healing rates were 13.82%±0.85% in blank control group and 13.28%±0.86 % in negative control group and 27.43%±0.80% in MAP4K1 overexpression group and 9.94% ± 0.85% in MAP4K1 inhibition group respectively.There were significant difference among MAP4K1 overexpression group and blank control group and negative control group,(P< 0.05)and also among MAP4K1 inhibition group and blank control group and negative control group,(P< 0.05).Scratched 72 hours later,the scratch healing rates were 17.24%±0.80% in blank control group and 16.34%±0.90% in negative control group and 29.69% ± 0.78% in overexpression group and 11.35%±0.43% in MAP4K1 inhibition group respectively.There were significant difference among MAP4K1 overexpression group and blank control group and negative control group,(P< 0.05)and also among MAP4K1 inhibition group and blank control group,negative control group,(P< 0.05).Conclusion : MAP4K1 can promote proliferation,migration and invasion of T24 cell,suggesting that there is a close relationship between MAP4K1 and bladder urothelial carcinoma.