| Objective:This study aims to investigate expressions of SATB1and EMT markers in bladder transitional cell carcinoma (BTCC) tissue as well as the relationships between SATB1and EMT markers and further evaluate the regulating mechanisms of SATB1-inducing EMT. Methods:Immunohistochemisty, Western blotting and qRT-PCR analysis were used to detect the expression of SATB1, E-cadherin (epithelial markers), vimentin (mesenchymal markers) in BTCC tissues and adjacent noncancerous tissues. For each specimen, the cancerous tissue and its remote normal mucosa were analyzed and compared. Pearson’s chi-squared (χ2) test or Fisher’s exact test was used to analyze the significance of correlations between SATB1expression and clinicopathological features of bladder cancer. Kaplan-Meier plots were used to estimate the prognostic relevance of SATB1in a univariate analysis. Multivariate analysis was performed using a COX proportional hazards test. Cell invasion and migration, cell cycle, cell proliferation and apoptosis were evaluated in SATB1knockdown and overexpressed cell lines.Results:Our results showed that the expression of SATB1, Snail, Slug and Vimentin ware remarkably up-regulated,but E-cadherin was down-regulated in BTCC tissues. Immunohistochemical staining results showed that the positive rate of SATB1, Snail, Slug, Vimentin and E-cadher were64.4%,7.2%,2.8%,71.2%,12.1%in BTCC cases, respectively. In addition, Pearson’s chi-squared test showed that SATB1overexpression was significantly associated with age (P=0.034), depth of invasion (P<0.001), TNM stage (P=0.015), lymph node status (P=0.013) and distant metastasis status (P=0.012). Further more, Spearman analysis indicated that the high level of SATB1was positively correlated with Snail, Slug expression, while significantly negative correlations with E-cadherin mRNA. In addition, patients with high SATB1expression had a shorter5-year survival compared with those with low SATB1 expression, SATB1was an independent risk factor for patients with bladder cancer.Conclusion:Our results suggest that SATB1is significantly associated with EMT-related markers and may play a crucial role in regulating EMT processes involved in carcinogenesis and progress of BTCC. Further, it may be a novel therapeutic target for aggressive bladder cancers. Objective:To observe the expression of SATB1and EMT regulated genes in different cancer cell lines, and to explore the regulation mechanism of SATB1-mediating EMT and its effect on biologic behavior of bladder cancer cell biology in proliferation, invasion ability.Methods:Real-time quantitative PCR, western blotting and immunofluorescence staining were used to examine the mRNA and protein levels of SATB1and EMT markers in bladder cancer cell lines of different invasiveness. To discuss the relationships between SATB1and EMT markers SATB1in bladder cancer cell lines of different invasiveness. The SATB1overexpression and SATB1gene silencing cell model were established and the differential expression of SATB1and EMT regulated genes, such as Snail, Slug, E-cadherin and Vimentin were tested and the cell morphological changes were observated in SATB1overexpression and silencing cell.We next performed the CCK-8and Transwell assays to investigate the effects of down-or up-regulation SATB1expression on biologic behavior of bladder cancer cell in proliferation, invasion ability in vitr, based on constructing SATB1-specific shRNA pGenesil2-SATBl and SATB1expression vector pcDNA3.1-SATB1.Results:the expression of SATB1, Snail, Slug and Vimentin levels in highly invasive bladder cancer cell T24were significantly higher than those in low invasive bladder cancer cell BIU-87(P<0.001), while the expression of E-cadherin was significantly lower than that in BIU-87cells; After the successful construction of pcDNA3.1-SATB1 overexpression and pGenesil2-S ATB1-shRNA interference plasmid, the SATB1overexpression cell model in low invasive bladder cancer cell BIU-87was established. In addition, BIU-87cells treated with pcDNA3.1-SATB1expression plasmids exhibited a marked change in morphology, from a cobblestone-shaped morphology to a spindle-shaped morphology and T24cells treated with pGenesil2-SATB1-shRNA expression plasmids exhibited major cell morphological changes, from a spindle-like fibroblastic morphology to a cobble-stone-like morphology by immunofluorescence test. Furthermore, the E-cadherin was found to be increased in the established pcDNA3.1-SATB1BIU-87cells, but increased on the plasma membrane in the established SATB1-shRNA T24cells. Western blotting and real-time quantitative PCR results showed that Snail, Slug and Vimentin expression were significantly increased in pcDNA3.1-SATB1group, while the E-cadherin was down regulated, and cell proliferation, invasive ability were enhanced obviously. In addition, the ratio of cells in G0/G1phase decreased, and the ratio of cells entering S phase increased significantly compared to the control group (P<0.05). There was no significant difference in apoptosis rate between groups (P>0.05); However, the levels of Snail, Slug and Vimentin expression were significantly down regulated in pGenesil2-SATB1-shRNA group cells, but the E-cadherin was upregulated significantly, and cell proliferation, invasion ability decreased significantly. The cell cycle progression was arrested in GO/G1phase, and the ratio of cells entering S phase was reduced significantly in T24cells treated with pGenesil2-SATB1-shRNA (P<0.05). Nevertheless, the apoptotic rate didn’t increase in SATB1-knockdown cells (P>0.05); Spearman rank analysis showed that there was a significant positive correlation between SATB1expression and Snail, Slug and Vimentin, but there was significantly negative correlation between SATB1and E-cadherin.Conclusion:SATB1promotes the proliferation, invasion and cell cycle of human bladder cancer cells, but no obvious effect on cell apoptosis. SATB1may promote the expression of Snail, Slug and inhibit the E-cadherin expression and further induce EMT occurs. SATB1and plays a key role in the progress of bladder cancer. |
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