Knockdown of SIRT1 Suppresses Bladder Cancer Cell Proliferation and Migration and Induces Cell Cycle Arrest and Antioxidant Response through FOXO3a-Mediated Pathways

Figure 5

Decreased SIRT1 could not lead to BCa cell apoptosis. (a) The human bladder cancer cells EJ (contaminated by T24 as per “http://iclac.org/databases/cross-contaminations/”) and T24 were transfected with SIRT1-target-specific-siRNA (B, D) and control-siRNA (A, C) for 48 h. BCa cells were stained with Annexin V/PI and apoptosis was measured by flow cytometry. (b) Statistical analysis of TUNEL-test revealed no significant (n.s.) increase of apoptotic cell rate in either EJ (contaminated by T24 as per “http://iclac.org/databases/cross-contaminations/”) or T24 cells after SIRT1-target-specific-siRNA (SIRT1 KD) treatment. (c) TUNEL-test to detect apoptotic cells (green) in control-siRNA-treated BCa cells (A, C) and SIRT1-target-specific-siRNA-treated BCa cells (B, D). Nuclei (blue) were stained by DAPI. (d, e, f) Representative immunofluorescence images of cleaved-Caspases 3, 7, and 9 (green) in the T24 and EJ (contaminated by T24 as per “http://iclac.org/databases/cross-contaminations/”) cells after SIRT1-target-specific-siRNA treatment (KD) (B, D), compared with control-siRNA treatment (NC) (A, C). Nuclei (blue) were stained by DAPI. The scale bars for (c–f) are 40 μm.

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