Fig. 6: PTEN-K260 deacetylation by SIRT7 promotes endometrial cancer metastasis.

A Wound healing assay in HEC-1B-rescue cells. For HEC-1B-rescue cells, PTEN was knocked down and empty vector, rPTEN-WT, rPTEN-K260Q or rPTEN-K260R was expressed. Relative area of migration was measured and normalized to the ‘EV’ group, n = 6, cell cultures from three independent experiments, scale bar = 500 μm. B Transwell and invasion assay in HEC-1B-rescue cells. Relative cell number was quantified, n = 8, cell cultures from four independent experiments, scale bar = 200 μm. C Wound healing assay in HEC-1B cells. SIRT7 was overexpressed and then PTEN-WT, PTEN-K260Q or empty vector was then expressed, n = 6, cell cultures from three independent experiments, scale bar = 500 μm. D Transwell and invasion assay in HEC-1B cells. SIRT7 was overexpressed and then PTEN-WT, PTEN-K260Q or empty vector was then expressed, n = 8, cell cultures from three independent experiments, scale bar = 200 μm. E Representative images of the mice liver, spleen, and H&E-stained liver after the injection of HEC-1B-rescue cells into the spleen. Tumor burden was calculated, EV group n = 12, rPTEN-K260Q/rPTEN-K260R group n = 10, scale bar = 100 μm, metastasis nodules marked with asterisks. F Representative images of the mice liver, uterus, and H&E-stained liver, n = 6, scale bar = 100 μm. The incidence of liver metastasis nodules was shown and tumor burden was calculated. G AcK260-PTEN level analysis of human EC tissue by IHC. Primary tumor and paired metastasis tissue (in ovary/fallopian, n = 13) or metastatic lymph nodes (n = 15) were stained with AcK260-PTEN and quantified by IHC score. Scale bar = 50 μm H AcK260-PTEN level analysis of human EC tissue by IHC. Tumor (n = 21) and paired para-tumor tissue (n = 18) were stained with AcK260-PTEN, and quantified by IHC score. Scale bar = 50 μm. The P value was calculated with two tailed unpaired t test for (A–F, H), and was calculated with two tailed paired t test for (G). I Kaplan–Meier overall survival analysis of EC patients with or without PTEN mutation, data from the TCGA, n = 175 for PTEN-WT (SIRT7 high = 117; SIRT7 low = 58), n = 332 for PTEN-mut (SIRT7 high = 222; SIRT7 low = 110). Cut-off = 0.33. Data are presented as mean values ± s.d. for (A–D), and ± s.e.m. for (E–H).