Figure 6.

SAHA treatment exerts anticancer activity through mediating Parkin acetylation. (A) HeLa cells with WT or mutant Parkin expression were subjected to SAHA treatment (5 μmol/L, 24 h) and cell morphology was captured using a Phase contrast microscopy. Scale bar = 100 μm. (B) as in (A), cells were seeded into 96-well plate and treated with SAHA. CCK-8 solution was added into each well of the plate and the absorbance was measured at 450 nm using microplate spectrophotometer. (C) as in (A), under SAHA treatment, cell proliferation was determined by colony formation assay. After crystal violet staining, the formed colonies were photographed. (D) as in (A), cells were seeded into 6-well plate and cultured for consecutive 5 days. Each day, cells were trpsined and counted and cell growth curve was drawn accordingly. (E) 2 × 10⁶ HeLa cells with empty vector, wild-type or mutant Parkin expression, was subcutaneously injected into the right flanks of the nude mice. Tumor-bearing mice were intraperitoneally injected with SAHA (50 mg/kg) or saline, respectively. Xenograft tumor volumes were measured twice per week and then quantified and statistically analyzed. (F) When mice execution, xenograft tumors from different groups were removed and typical images were captured. (G) Representative histopathological images of tumor sections were from hematoxylin and eosin staining. The expressions of Ki67, cleave-caspase3 and Parkin in tumor xenograft were detected using immunohistochemistry. The images were taken at 400 × magnification. (H) A schematic model of the acetylation of Parkin in mitophagy and tumor suppression by HDACis in human cervical cancer. ^#P > 0.05, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.