The Expression Of TACC3 In Bladder Urothelial Carcinoma And Its Influence To Migration And Invasion In T24 Cells

Objective:To detect the expression of TACC3 protein in urothelial carcinoma of bladder T24 cells and explore the influence of down-regulating TACC3 expression level on migration and invasion through epithelial-mesenchymal transition(EMT) in bladder cancer T24 cells. Methods:Using western blot to detect the expression level of TACC3 protein in bladder urothelial carcinoma T24 cells and SV-HUC-1 cells; In vitro studies,we design and build several shRNA expression plasmid and screen the most effective target by qPCR. Then use TACC3- shRNA related plasmid transfect bladder cancer T24 cell through cytology transfection technique to downgrade TACC3 level. The interference effect is detected by western blotting and qPCR, while detecting TACC3 and EMT related mRNA and protein expression level. Though cell migration test to measure repair rate of 24 h and transwell invasion test to verify its cell invasion. Result:In bladder urothelial carcinoma T24 cells and SV-HUC-1 cells, the result of western blot showed that the relative grey level are 0.22±0.02 ? 0.99±0.05,respectively. The expression level of TACC3 protein in T24 cells was obviously higher than that in SV-HUC-1 cells, the difference was statistically significant(P < 0.05). In vitro, successfully building four TACC3-shRNA interference plasmids,we screened TACC3 mRNA expression by qPCR to select a most efficient TACC3-shRNA. Its interference effect as high as 90.5%. down-regulating TACC3 level, qPCR technique was used to detect TACC3 and EMT related(E-cadherin, Twist,Vimentin, Snail and MMP-9) mRNA changes, the results shown: the relative mRNA expression of EMT markers(E-cadherin, Twist, Vimentin, Snail and MMP-9) were 1.11±0.13?0.26±0.04?0.38±0.06?1.11±0.04?0.41±0.03; Western Blot detected target protein expression level above, the results show that TACC3 protein, EMT related(E-cadherin, Twist, Vimentin, Snail and MMP-9) protein relative grey value were 1.08±0.06?0.17±0.04?0.07±0.01?0.14±0.01?0.35±0.13,respectively. Result of scratch test showed repair rate of T24-TACC3-shRNA was(25.50±0.9609)%(24h), transwell invasion test showed under the average visual field, the average cell numbers of the control group. NC group and shRNA group were 326.8±14.09, 311.3±8.664 and 143.3±10.62 respectively, which were statistically significant. Conclusion:TACC3 is highly expressed in bladder urothelial carcinoma T24 cells. In vitro study, we demenstrated that on T24 bladder cancer cells induced with the occurrence of EMT signaling, down-regulating TACC3 by importing the interference of efficiency for 90.5% of the pYr-LVX-TACC3-shRNA plasmids can inhibit migration and invasion ability of T24 bladder cancer cells. The result shows TACC3 protein must act a crucial role in the migration and invasion of bladder urothelial cancer through adjusting the EMT signaling. TACC3 would be an important biomarker for bladder cancer targeted therapy, and providing a new way for the diagnosis and treatment of bladder cancer.