| Objective:To explore the expression of SATB1in human renal cell carcinoma (RCC) cell lines and the effects of SATB1expression on cell proliferation and invasion of RCC cells.Methods:The mRNA and protein levels of SATB1in RCC cell lines were examined by real-time quantitative PCR, western blotting and immunofluorescence staining, respectively. We next performed the CCK-8and Transwell assays to investigate the effects of down-or up-regulation SATB1expression on cell proliferation and invasion of RCC cells in vitro, respectively, based on constructing SATB1-specific shRNA pGenesil2-SATB1and SATB1expression vector pcDNA3.1-SATB1.Results:SATB1mRNA and protein levels were dramatically increased in human RCC cell lines as compared with the immortalized normal human proximal tubule epithelial cell line HK-2(P<0.001for both), and levels of SATB1expression increased sequentially in RCC cell line ACHN, A498and786-O. As compared with control groups, both mRNA and protein levels of SATB1were significantly decreased in786-O cells transfected with pGenesil2-SATB1-shRNA (P<0.001), whereas SATB1expression levels remarkably increased in ACHN cells transfected with pcDNA3.1-SATB1(P<0.001). Evaluations of cell proliferation by CCK-8assays showed that down-regulation of SATB1dramatically inhibited the cell proliferation rates of786-O cells, while overexpression of SATB1significantly promoted the growth of ACHN cells (P<0.05for both). Furthermore, Transwell assays showed that down-regulation of SATB1in786-0cells transfected with SATB1shRNA resulted in a significant inhibition of cell invasion as compared with that in the control groups (P<0.001), whereas SATB1over-expression significantly promoted the invasion of ACHN cells stably transfected with pcDNA3.1-SATB1(P<0.001). Conclusion:SATB1expression was significantly up-regulated in RCC cells. Over-expression of SATB1may play critical roles in the acquisition of an aggressive phenotype for RCC cells and the development of invasion and metastasis of RCC, suggesting that SATB1is a promising biomarker and therapeutic target for patients with RCC. Objective: To investigate expressions of SATB1and EMT markers in human clear cell renal cell carcinoma (ccRCC) tissue as well as the relationships between SATB1and EMT markers.Methods:Immunohistochemical staining, western blotting and real-time quantitative PCR were employed to examine SATB1expression in specimens from89cases of ccRCC. Expressions of SATB2and EMT markers (including E-cadherin, ZEB2, vimentin and fibronectin) in ccRCC tissue were studied by immunohistochemistry, and relationships between SATB1and expressions of SATB2and EMT markers were statistically analyzed by Spearman rank correlation analysis.Results:Immunohistochemical staining showed that SATB1, SATB2, E-cadherin, ZEB2, vimentin and fibronectin were positively expressed in46.1%,42.7%,44.9%,51.7%,37.1%and49.4%of ccRCC cases, respectively. As compared with paired normal tissues, the levels of SATB2and E-cadherin were significantly decreased in ccRCC tissue, whereas expressions of SATB1and ZEB2were dramatically up-regulated. In addition, over-expression of SATB1was significantly associated with depth of invasion (P<0.001), TNM stage (P=0.009) and lymph node status (P=0.001). Further Spearman analysis indicated that the high level of SATB1was positively correlated with ZEB2expression (R=0.262, P=0.013), while significantly inverse correlations were observed between SATB1level and expressions of SATB2and E-cadherin proteins (R=-0.296, P=0.005; R=-0.427, P<0.001), respectively. Conclusion:Our results indicated that SATB1expression was remarkably correlated with levels of EMT markers in ccRCC tissue. Combined analysis of expressions of SATB1and EMT markers in ccRCC tissue might have significant values to further investigate roles of SATB1and EMT which played in determining invasion and metastasis of renal cell carcinoma (RCC), and assessing prognosis of patients with RCC. |
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