Fig 1: TPX2 expression and function in prostate tumor cell lines. (A) Western blotting detected TPX2 protein expression in PCa cell lines. TPX2 was highly expressed in the DU145 cell line, while expressed at low levels in the PC3 and LNCaP cell lines. The CCK8 assay indicated that the overexpression of TPX2 in (B) LNCaP and (C) PC3 cells promoted the proliferation rate of PCa cells compared with the control group. The flow cytometry assay conducted in (D) LNCaP-NC and (E) LNCaP-TPX2 cells suggested that TPX2 inhibited cell cycle transit from G0/G1 to S/M phase in LNCaP-TPX2 cells. (F) The average proportion of cells in S+G2 phase was 40.607±2.172 in the LNCaP-TPX2 group, while it was 47.09±1.804 in the LNCaP-NC group (P=0.016). (G) PC3-NC and (H) PC3-TPX2 cells were also examined using flow cytometry. (I) No significant difference in PC3 cells was observed (P>0.05). TPX2, Targeting protein for Xenopus kinesin-like protein 2; PCa, prostate cancer; NC, negative control; NS, not significant. *P<0.05.
Fig 2: Overexpression of TPX2 inhibits apoptosis in LNCaP and PC3 cells. Cells overexpressing TPX2 were stained using Annexin V, and it was demonstrated that TPX2 decreased the rate of apoptosis in (A) LNCaP-NC and (B) LNCaP-TPX2 cells. (C) The apoptotic rate in the LNCaP-TPX2 group was 2.83±0.42, while it was 4.25±0.13 in the LNCaP-NC group (P=0.005). No difference in the apoptotic rate was observed between (D) PC3-NC and (E) PC3-TPX2 cells treated with docetaxel. (F) The apoptosis rates in the PC3-TPX2 and PC3-NC groups were similar (36.92±1.49 vs. 38.49±2.36; P=0.383). TPX2, Targeting protein for Xenopus kinesin-like protein 2; PCa, prostate cancer; NC, negative control; NS, not significant. *P<0.05.
Fig 3: Immunohistochemical staining for TPX2 protein expression in PCa and adjacent benign tissues. (A) The whole scanned image of the tissue array (B) Statistical analyses indicated an increased immunoreactivity score in cancer tissues compared with benign tissues. (C) Enlarged image of BPH tissue indicated that TPX2 was only weakly expressed in benign tissues. Red box demonstrates the blue box at a magnification. (D) Enlarged image of PCa tissue demonstrated that TPX2 was highly abundant in the nucleus and cytoplasm of tumor cells. Red box demonstrates the blue box at a magnification. TPX2, Targeting protein for Xenopus kinesin-like protein 2; PCa, prostate cancer; BPH, benign prostatic hypertrophy. *P<0.05.
Fig 4: High TPX2 expression is associated with poor prognosis in patients with PCa. (A) Kaplan-Meier analysis of the BCR time of patients with PCa with high TPX2 expression levels was shorter compared with those with low TPX2 expression levels (P=0.007). (B) The overall survival time of patients with PCa was not associated with TPX2 expression levels (P=0.120). TPX2, Targeting protein for Xenopus kinesin-like protein 2; PCa, prostate cancer; BCR, biochemical recurrence-free time.
Fig 5: Overexpression of TPX2 enhanced wound healing of LNCaP and PC3 cells. Wound healing assay analysis was performed at 0, 24 and 48 h with or without TPX2 overexpression. The migratory ability was increased in (A) LNCaP-TPX2 and (B) PC3-TPX2 cells at 24 and 48 h. TPX2, Targeting protein for Xenopus kinesin-like protein 2; NC, negative control. **P<0.01.
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