Expression Of MAP4K1 In Bladder Urothelial Carcinoma And Its Relationship With JNK Signaling Pathway

Objective:To study the mechanism of MAP4K1 and the relationship with its downstream JNK signaling pathway in bladder urothelial carcinoma the expression of the taget gene MAP4K1 of miR-96 was detected and in order to provide theoretical results for the clinical diagnosis,prognosis and gene therapy of bladder urothelial carcinoma.Materials and methods:?1?The expression of the target gene MAP4K1 in bladder urothelial carcinoma tissue was detected and measured the optical density by immunohistochemical staining method through the tissue microarray.?2?The mRNA expression levels of MAP4K1?JNK?C-jun in T24cells transfected with mi R-96 inhibits was detected by using real-time fluorescence quantitative PCR method.The experimental group was:blank control group?T24 cytome??negative control group?T24+transfected with mi R-96 negative control?and experimental group?T24+transfected miR-96 inhibits agent group?,cultured T24 cells,using Lipofectamine^TM 2000 to synthetic miR-96 inhibitors in T24 cells transfected with in vitro.After 48 hours of cell culture,RNA was extracted and was detected.?3?The protein expression levels of MAP4K1?JNK and C-jun in T24cells transfected with miR-96 inhibits was detected by using western blot method.The experimental group was:blank control group?T24 cytome??negative control group?T24+transfected with miR-96 negative control?and experimental group?T24+transfected mi R-96 inhibits agent group?,cultured T24 cells,using Lipofectamine^TM 2000 to synthetic miR-96inhibitors in T24 cells transfected with in vitro.After 48 hours of cell culture,total protein was extracted and detected by western blot.Results:?1?The results of immunohistochemistry showed that MAP4K1 was mainly expressed in cytoplasm and nucleus.The optical density in bladder cancer tissue was 0.24±0.04,and the optical density in normal bladder tissue was 0.16±0.02,the difference between the two groups was significant?p<0.05?.?2?RT-PCR results showed that the mRNA relative expression of MAP4K1 in blank group,control and experimental group was 2.51±0.362.50±0.48,1.03±0.11,The statistical differences was significantly in the experimental group while compared with the blank group and the negative control group?p<0.05?.The mRNA relative expression of JNK was 1.88±0.14,1.90±0.36 and 1.05±0.18,The statistical differences was significantly in the experimental group while compared with the blank group and the negative control group?p<0.05?.The mRNA relative expression of C-jun was 1.17±0.19,1.12±0.17and 1.03±0.12,The statistical differenceswas significantly in the experimental group while compared with the blank group and the negative control group?p<0.05?.?3?The results showed that the expression of MAP4K1 protein in the experimental group was significantly lower than the control group and blank group,and the expression of JNK?C-jun in the downstream of JNK signal pathway was significantly lower than the control group and blank group?p<0.05?.?4?The experimental data are expressed with x±s and use graphPad prism and SPSS19.0 statistical analysis software to analyze.The comparison of the experimental group and control group will employ T test,and more than two groups of data comparison use analysis of variance.Conclusion:1.MAP4K1 was overexpressed in the bladder urothelial carcinoma tissue,and which was positive correlation with mi R-96.2.When the expression of mi R-96 was inhibited,the expression of MAP4K1 and JNK and C-jun mRNA and protein in the downstream of the signal pathway was also inhibited.