Fig 2: GOLPH3 overexpression induces metastasis in colon cancer in vivo.A Zebrafish models injected with HCT-116-PMSCV-Vector and HCT-116-PMSCV-GOLPH3 cells were used. Green fluorescence indicates zebrafish, and red fluorescence was used to mark the cell lines. At 1-week post-injection, the disseminated tumor foci count was significantly higher in the GOLPH3-overexpression model (P = 0.0037). B HCT-116-pSuper-Vector and HCT-116-pSuper-GOLPH3 sh1# cells were also used to establish a zebrafish model. In contrast to A, the disseminated tumor foci count was significantly lower in the GOLPH3-silencing model (P = 0.0004).

Fig 3: Kaplan-Meier curves for FAP-a and GOLPH3 state and duration for recurrence among DCIS patients. (Left panel) The FAP-a+GOLPH3+ immunophenotype is correlated with the induction of overall recurrence. (Middle panel) The FAP-a+GOLPH3+ immunophenotype is associated with the increase of DCIS recurrence. (Right panel) The FAP-a+GOLPH3+ immunophenotype is correlated with the induction of DCIS progression to IDC. (FAP-a+GOLPH3+ immunophenotype, score = 1; FAP-a-GOLPH3-, score = 2). p-values (log rank test) are as shown.

Fig 4: GOLPH3 facilitates autophagy via the suppression of phosphorylation of Akt at Ser473.A KEGG (Kyoto Encyclopedia of Genes and Genomes) analyses differentially expressed genes between the HCT-116-PMSCE-Vector and HCT-116-PMSCV-GOLPH3 cells. B Western blot analysis of GOLPH3-overexpressing and control HCT-116 and HT-29 cell lines. Akt Ser473 phosphorylation was decreased in GOLPH3-overexpressing cells. In GOLPH3-silenced colorectal cancer cells, Akt Ser473 phosphorylation was enhanced. C MK-2206 2HCl was used to repress Akt phosphorylation. P62 and E-cadherin were stimulated, and N-cadherin was repressed when GOLPH3 was silenced. When CRC cell lines were cultured in Akt Ser473 phosphorylation inhibitor, P62 and E-cadherin were repressed, and N-cadherin was enhanced. GAPDH was used as an internal reference. D The above results demonstrated that GOLPH3 induces autophagy and EMT to promote metastasis in colon cancer, via the repression of Akt Ser473 phosphorylation.

Fig 5: GOLPH3 regulated 5-FU-induced apoptosis via modulation of PARP cleavage. RKO and LoVo cells were transfected with pCMV-Myc-GOLPH3/pCMV-Myc (GOLPH3 over-expression, A and C) or GOLPH3 siRNA pool (siG-pool)/control siRNA (siControl) (GOLPH3 silencing, B and D) and then treated with indicated concentrations of 5-FU (0, 100, 200 µM) for 48 h. Western blot analysis showed the alteration of cleaved PARP and GOLPH3. Upper panels, one representative of three similar Western blot analysis. Lower panels, relative protein ratio were normalized to ß-actin. All data are mean ± SD, *P < 0.05.