Fig. 4

ASPA suppresses the tumorigenic behavior of PCa cells in vivo. A Representative images of subcutaneous xenograft tumors derived from PC-3 cells transfected with control or ASPA overexpression vector. b The volume of xenograft tumors in the control group and ASPA overexpression group (n = 8). c The weight of xenograft tumors in the control group and ASPA overexpression group (n = 8). d Representative images of IHC showed the expression of Ki67 in xenograft tumors in the control group and ASPA overexpression group (scale bar = 100 μm). e RT-qPCR results of proliferation-related genes and epithelial–mesenchymal transition genes in xenograft tumors in the control group and ASPA overexpression group. The mRNA expression levels were normalized to ACTB levels. f Western blotting results (left) and quantification (right) results for ASPA, proliferation-related proteins, and epithelial–mesenchymal transition proteins in xenograft tumors in the control group and ASPA overexpression group. Protein expression levels were normalized to β-actin levels. g Representative images of subcutaneous xenograft tumors derived from PC-3 cells transfected with shRNA and shASPA. h The volume of xenograft tumors in the shRNA group and shASPA group (n = 8). i The weight of xenograft tumors in the shRNA group and shASPA group (n = 8). j Representative images of IHC showed the expression of Ki67 in xenograft tumors in the shRNA group and shASPA group (scale bar = 100 μm). k RT-qPCR results of proliferation-related genes and epithelial–mesenchymal transition genes in xenograft tumors in the shRNA group and shASPA group. The mRNA expression levels were normalized to ACTB levels. l Western blotting results (left) and quantification (right) results for ASPA, proliferation-related proteins, and epithelial–mesenchymal transition proteins in xenograft tumors in the shRNA group and shASPA group. Protein expression levels were normalized to β-actin levels. m Representative images of orthotopic xenograft tumors derived from PC-3 cells transfected with control or ASPA overexpression vector. n The weight of orthotopic xenograft tumors in the control group and ASPA overexpression group (n = 8). o Western blotting results (left) and quantification (right) results for ASPA, proliferation-related proteins, and epithelial–mesenchymal transition proteins in orthotopic xenograft tumors in the control group and ASPA overexpression group. Protein expression levels were normalized to β-actin levels. p Representative images of orthotopic xenograft tumors derived from PC-3 cells transfected with shRNA or shASPA. q The weight of orthotopic xenograft tumors in the shRNA group and shASPA group (n = 8). r Western blotting results (left) and quantification (right) results for ASPA, proliferation-related proteins, and epithelial–mesenchymal transition proteins in orthotopic xenograft tumors in the shRNA group and shASPA group. Protein expression levels were normalized to β-actin levels. The data are presented as the mean ± standard deviation (SD). ASPA aspartoacylase, CCND1 cyclin D1, CDH1 Cadherin 1, CDH2 Cadherin 2, IHC immunohistochemistry, PCa prostate cancer, MYC v-Myc myelocytomatosis viral oncogene homolog, MMP9 matrix metallopeptidase 9, PCNA proliferating cell nuclear antigen, RT-qPCR real-time quantitative PCR, shRNA small hairpin RNA. *P < 0.05, **P < 0.01