Fig 1: SKA3 regulates epithelial-mesenchymal transition (EMT), MAPK/ERK pathway in vitro and regulates the aggressive behaviors in vivo. (A) Silenced for SKA3 inhibits proliferation, EMT and MAPK/ERK related proteins in MGC-803 and HGC-27. (B) Tumor growth assessment in subcutaneous tumor model of gastric cancer. (C) Tumor weight was measured in shSKA3 and control groups. (D) Tumor volume was measured in shSKA3 and control groups. (E) Intraperitoneal metastasis model showed that the metastatic nodules derived from shSKA3 and control groups. (F) Intraperitoneal metastasis tumor weight was measured in shSKA3 and control groups. (G) Microarray heatmap showed differentially expressed genes. (H) Differentially expressed genes were screened by qRT-PCR in MGC-803. (I) Differentially expressed genes were screened by qRT-PCR in HGC-27. (J) Western blot of DUSP2 levels in the sh-SKA3 cells. Data are shown as mean ± SD for triplicate experiments. The two-tailed Student’s unpaired t-test was used. *p < 0.05, **p < 0.01, ***p < 0.001.
Fig 2: SKA3 promotes proliferation in gastric cancer. (A) mRNA and protein levels of SKA3 after transfection with shSKA3 lentivirus in MGC-803 and HGC-27. (B) CCK-8 cell proliferation assay detects cell viability by silenced for SKA3 in MGC-803 and HGC-27. (C) Cloning experiment detects cell growth by silenced for SKA3 in MGC-803 and HGC-27. (D) Flow cytometry detects cell cycle n by silenced for SKA3 in MGC-803 and HGC-27. Data are shown as mean ± SD for triplicate experiments. The two-tailed Student’s unpaired t-test was used. *p < 0.05, **p < 0.01, ***p < 0.001.
Fig 3: SKA3 promotes migration, invasion and adhesion in gastric cancer. (A) Wound-healing experiments detects cell migration by silenced for SKA3 in MGC-803 and HGC-27. (B) Transwell experiment detects cell migration by silenced for SKA3 in MGC-803 and HGC-27. (C) Transwell experiment detects cell invasion by silenced for SKA3 in MGC-803 and HGC-27. (D) Adhesion experiment detects cell adhesion by silenced for SKA3 in MGC-803 and HGC-27. (E) Relative healing rate in (A) were quantified and shown. (F) Relative migration numbers in (B) were quantified and shown. (G) Relative invasion numbers in (C) were quantified and shown. (H) Relative adhesion numbers in (D) were quantified and shown. Data are shown as mean ± SD for triplicate experiments. The two-tailed Student’s unpaired t-test was used. **p < 0.01, ***p < 0.001.
Fig 4: SKA3 is upregulated in gastric cancer. (A) SKA3 mRNA expression in TCGA database. (B) SKA3 mRNA expression in GEO database. (C) SKA3 mRNA expression in gastric cancer tissues. (D) SKA3 protein levels in gastric cancer tissues. N, normal tissue. T, tumor tissue. (E) SKA3 mRNA expression in gastric cancer cell lines. (F) SKA3 protein levels in gastric cancer cell lines. (G) SKA3 and DUSP2 expression in gastric cancer tissues, normal tissues, peritoneal tissues by immunohistochemistry (IHC). (H) The correlation between SKA3 and DUSP2 IHC score by scatter plots. (I) Kaplan-Meier survival curves about SKA3. (J) Kaplan-Meier survival curves about DUSP2. (K) The prognostic significance of SKA3 high and low expression in Kaplan-Meier plotter database.*p < 0.05, **p < 0.01, ***p < 0.001.
Fig 5: DUSP2 inhibits proliferation, migration, invasion and adhesion in gastric cancer. (A) CCK-8 cell proliferation assay detects cell viability by overexpression for DUSP2 in MGC-803 and HGC-27. (B) Cloning experiment detects cell growth by overexpression for DUSP2 in MGC-803 and HGC-27. (C) Flow cytometry detects cell cycle by overexpression for DUSP2 in MGC-803 and HGC-27. (D) Wound-healing experiments detects cell migration by overexpression for DUSP2 in MGC-803 and HGC-27. (E) Transwell experiment detects cell migration by overexpression for SKA3 in MGC-803 and HGC-27. (F) Transwell experiment detects cell invasion by overexpression for SKA3 in MGC-803 and HGC-27. (G) Adhesion experiment detects cell adhesion by overexpression for SKA3 in MGC-803 and HGC-27. Data are shown as mean ± SD for triplicate experiments. The two-tailed Student’s unpaired t-test was used. *p < 0.05, **p < 0.01, ***p < 0.001.
Supplier Page from Abcam for Anti-SKA3 antibody