Fig. 5: The effect of CsA on PPIL2 and EMT in breast cancer cells | Cell Death & Disease

Fig. 5: The effect of CsA on PPIL2 and EMT in breast cancer cells

From: U-box ubiquitin ligase PPIL2 suppresses breast cancer invasion and metastasis by altering cell morphology and promoting SNAI1 ubiquitination and degradation

Fig. 5

a Concentrations of 2 and 20 μg/ml CsA were added into the medium 16 h after the ZR-75-30 cells were plated. DMSO was taken as negative control. The cells were collected 12 h later and prepared for western blot. The expression of PPIL2, SNAI1, and SNAI2 was detected. b–d A concentration of 20 μg/ml CsA was added 16 h after the ZR-75-30 cells were plated. The cell lysate was collected at 4, 8, 12, and 16 h, respectively. The result of western blot showed that PPIL2 level increased in a time-dependent manner with CsA treatment within 16 h. The bar graphs showed the relative level of PPIL2 and SNAI1 in three independent experiments. *P < 0.05, **P < 0.01 e CsA raised PPIL2 abundance both in the cytoplasm and the nucleus in ZR-75-30 cells. f CsA prolonged the half-period of PPIL2 in ZR-75-30 cells. g The expression of EMT markers was examined in ZR-75-30 cells treated with 2 and 20 μg/ml CsA. h The level of SNAI1 ubiquitination was upregulated with CsA treatment, but PPIL2 silence attenuated the effect of CsA. i CsA induced F-actin arrangement in ZR-75-30 cells. Segments of F-actin fiber evenly distributed throughout the cell. The fibrous construction was reconstructed and prolonged after PPIL2 was silenced (40×). j, k the migratory and invasive ability of ZR-75-30 cells treated with 20 μg/ml was dampened compared to cells treated with DMSO (20×). The effect of CsA on cell motility was negated with siPPIL2 transfection. The bar graphs show the number of migrating and invading cells for each category of cells. Each bar represents the mean ± S.D. from three independent experiments. **P < 0.01

Back to article page