Fig 2: TCTP is secreted under low serum and hypoxic conditionsA. Extracellular TCTP secreted from different medium-conditioned CRC cell lines was measured by ELISA. *P<0.05, **P<0.01, ***P<0.001 by One-Way ANOVA. B. Western blot analysis of the expression levels of extracellular TCTP, intracellular TCTP, and TSAP6 in LoVo cells under serum free plus hypoxic conditions. C. Expression of intracellular TCTP under serum free and hypoxic conditions at the indicated time points was observed under a confocal microscope. Representative photographs are shown. TCTP, FITC dye (green); F-actin, Rhodamine B dye (red); nuclear, DAPI dye (blue). Scale bar: 40 μm. D. Expression and secretion of TCTP mediated by silencing of TSAP6 under serum free plus hypoxic conditions. E. HIF-1α (Δ401-603) promotes TCTP expression under normal conditions. F. 2-MeOE2 inhibits TCTP expression and secretion under serum free plus hypoxic conditions. G. Relative luciferase activity was measured by transfection of either pENTER, pGL6-pTCTP-luc and pRLTK or pENTER-HIF-1α (Δ401-603), pGL6-pTCTP-luc and pRLTK for 24h under normal conditions. F-luc: Firefly luciferase. R-luc: Renilla luciferase. NC: negative control.

Fig 3: Increased abundance of TPT1 protein in the tissue and sera of cervical cancer compared to cervicitis, CINIII, and controls. (A) Tissue IHC staining of TPT1 protein in the control and different cervical lesions including cervicitis, CINIII, and cervical cancer. Scale bars: x200, 50 μm; x400: 10 μm. (B) Serum TPT1 concentration in CINIII and cervical cancer patients, compared to normal healthy controls, measured by ELISA. IHC images are representative of all samples tested. *p < 0.05; NS: statistically not significant.

Fig 4: GSEA enrichment analysis of TCGA data on gene sets and pathways associated with low (A) or high (B) TPT1 expression in cervical cancer. The horizontal bar, gradient-filled from red to blue, represents where genes in each gene set appear in the ranked list of genes. Genes on the left side (red) correlate most strongly with the phenotype. The vertical black lines represent the running enrichment scores as the projection of individual genes onto the horizontal ranked gene list. The bottom ranking matric in gray, moving from above zero (positively correlated) to below zero (negatively correlated), measures a gene’s correlation with the phenotype profile. Statistical variables of GSEA analysis are displayed in each image. ES, enrichment score for the gene set, reflecting the degree to which this gene set is over-represented at the peak (furthest from 0.0) of the entire ranked list of genes; p-value, statistical assessment of the significance against null distribution; NES, normalized enrichment score calculated by adjusting ES for gene set size or multiple hypothesis testing across analyzed gene sets; FDR (false discovery rate q-value), the estimated probability that a given NES represents a false positive finding. p < 0.05 and FDR < 0.25 were considered statistically significant.

Fig 5: Schematic of ischemia and hypoxia-induced TCTP secretion and the role of extracellular TCTP in promoting CRC cell invasiveness and metastasis