Fig 2: The effect of PRAME on tumor growth via Wnt/ß-catenin pathway in vivo. (A) The image of xenografts injected with PRAME-knockdown SiHa cells and control SiHa cells. (B) The weight of the xenografts in PRAME-knockdown group and control group. (C) The growth curves of xenograft tumors of PRAME-knockdown group and control group. (D) The immunofluorescence staining of Ki67 in tumor slices. (E) The immunofluorescence staining of TUNEL assay in tumor slices. (F) The image of xenografts injected with PRAME-overexpressed C33A cells and control C33A cells after the MSAB therapy. (Tumor groups as G1: Control cDNA+DMSO, G2: PRAME cDNA+DMSO, G3: Control cDNA+MSAB, and G4: PRAME cDNA+MSAB). (G) The weight of the xenografts in PRAME-overexpressed group and control group with or without MSAB therapy. (H) The growth curves of the xenografts in PRAME-overexpressed group and control group with or without MSAB therapy. (I) The TUNEL and IHC staining of tumor tissues. * p < 0.05, ** p < 0.01, *** p < 0.001.

Fig 3: Regulation of EMT-related proteins and Wnt/β-catenin pathway by PRAME expression in cervical cancer cells. (A) The change in E-cadherin and N-cadherin expression in PRAME-knockdown SiHa and PRAME-overexpressed C33A cells. (B) Western blotting analysis of Wnt/β-catenin signaling pathway-related protein expression after PRAME knockdown in SiHa cells and PRAME overexpression in C33A cells. (C) Western blotting analysis of Wnt3a and β-catenin expression in PRAME-overexpressed C33A and SiHa cells after MSAB treatment. The results of gray value quantification are displayed below the corresponding bands. (D) The cell viability in PRAME-overexpressed and control C33A and SiHa cells with MSAB treatment. (E) Left panel: The cell apoptosis in PRAME-overexpressed and control C33A and SiHa cells with MSAB treatment. Right panel: The quantitative results. *** p < 0.001.

Fig 4: Differential expression of PRAME in cervical cancer and normal cervix, and the role of PRAME expression in cell viability and proliferation in cervical cancer cells. (A) The expression of PRAME protein in adjacent normal tissues and cervical cancer tissues as shown by IHC staining. (B) The IHC score of PRAME protein. (Adjacent normal tissues: 22 cases, Cervical cancer tissues: 22 cases). (C) The expression of PRAME mRNA in normal cervical epithelium cells and cervical cancer cells as shown by qRT-PCR. (D) The expression of PRAME protein in normal cervical epithelium cells and cervical cancer cells as shown by Western blotting. (E) The quantitative analysis of (D). (F) PRAME-silenced C33A and SiHa cells were constructed via the two shPRAME sequences and confirmed by Western blotting. (G) The effect of PRAME knockdown on cell viability in C33A and SiHa cells was detected via CCK-8 assay. (H) The effect of PRAME knockdown on cell proliferation in C33A and SiHa cells was measured via BrdU assay. (I) The quantitative results of (H). (J) PRAME-overexpressed C33A and SiHa cells were constructed and confirmed via Western blotting. (K) The role of PRAME overexpression in cell viability in C33A and SiHa cells was detected via CCK-8 assay. (L) The role of PRAME overexpression on cell proliferation in C33A and SiHa cells was measured via BrdU assay. (M) The quantitative results for (L). * p < 0.05, ** p < 0.01, *** p < 0.001.

Fig 5: The role of PRAME expression on cell migration and invasion (A,B). The wound-healing rate in PRAME-silenced C33A and SiHa cells. (C,D) The quantitative results for (A,B). (E,F) The wound-healing rate in PRAME-overexpressed C33A and SiHa cells. (G,H) The quantitative results for (E,F). (I) The number of migratory and invasive cells in PRAME-knockdown SiHa cells. (J) The quantitative results for (I). (K) The number of migratory and invasive cells in PRAME-overexpressed SiHa cells. (L) The quantitative results for (K). * p < 0.05, *** p < 0.001.