The oncogenic role of Wnt10a in colorectal cancer through activation of canonical Wnt/β-catenin signaling - PubMed (original) (raw)
The oncogenic role of Wnt10a in colorectal cancer through activation of canonical Wnt/β-catenin signaling
Jinlong Li et al. Oncol Lett. 2019 Apr.
Abstract
Colorectal cancer (CRC) is one of the major causes of cancer-associated mortality worldwide. Wnt family member 10A (Wnt10a) is an oncogene associated with the carcinogenesis and progression of renal cell carcinoma, and is strongly expressed in the CRC cell line SW480. However, the role of Wnt10a in CRC has been rarely reported. In the present study, the expression levels of Wnt10a were higher in 40 tumor tissues compared with in paired control tissues, as determined by RT-qPCR method. In addition, the clinic opathological association analysis indicated that Wnt10a expression was associated with tumor stage (T3+T4, P=0.015). Furthermore, Wnt10a was highly expressed in the SW480, SW620 and HCT116 cell lines. In order to explore the role of Wnt10a in CRC, Wnt10a expression was knocked down by siRNA technology in HCT116 cell line. Cell proliferation was significantly inhibited by 55% in CCK-8 assay following Wnt10a knockdown and cell migration rate was decreased by 50% in Transwell assay. In addition, western blot analysis demonstrated that Wnt10a knockdown decreased the expression levels of β-catenin, cyclin D1, lymphoid enhancer-binding factor 1 and protein kinase B, which was consistent with results obtained with the Wnt/β-catenin specific inhibitor LGK-974. It was thus suggested that Wnt10a downregulation inactivated the Wnt/β-catenin signaling pathway in HCT116 cells. In conclusion, the present study demonstrated that Wnt10a may have an oncogenic role during carcinogenesis of CRC through activation of Wnt/β-catenin signaling.
Keywords: Wnt/β-catenin signaling; Wnt10a; migration; proliferation; siRNA.
Figures
Figure 1.
Expression levels of Wnt10a in colorectal tumor tissues and its clinical significance. (A) Wnt10a expression was significantly higher in the 40 tumor tissues compared with in the paired paratumoral tissues, as determined by reverse transcription-quantitative polymerase chain reaction. (B) Overall survival rate of patients with high Wnt10a expression was lower than in those with low Wnt10a expression. *P<0.05. Wnt10a, Wnt family member 10A.
Figure 2.
Effects of Wnt10a knockdown on HCT116 cell proliferation. (A) Expression levels of Wnt10a in three CRC cell lines: SW480, SW620 and HCT116. (B) Knockdown efficiency of Wnt10a in HCT116 cells assessed by reverse transcription-quantitative polymerase chain reaction. (C) Growth curve analysis of HCT116 cells transfected with Wnt10a-siRNA. *P<0.05. All data were obtained from triplicate experiments. NC, negative control; siRNA, small interfering RNA; Wnt10a, Wnt family member 10A.
Figure 3.
Effects of Wnt10a knockdown on HCT116 cell migration. (A) Migrated distance of HCT116 cells transfected with Wnt10a-siRNA1 within 24 h, as determined using the wound healing assay. Magnification, ×20. (B) Migration rate calculated from the wound healing assay. *P<0.05. All data were obtained from triplicate experiments. NC, negative control; siRNA, small interfering RNA; Wnt10a, Wnt family member 10A.
Figure 4.
Wnt10A knockdown suppresses tumor growth in vivo. (A) Expression levels of Wnt10a were decreased by Wnt10a-shRNA. (B) Tumor growth was delayed in mice treated with Wnt10a-shRNA. *P<0.05. NC, negative control; shRNA, short hairpin RNA; Wnt10a, Wnt family member 10A.
Figure 5.
Wnt10a knockdown inactivates the Wnt/β-catenin signaling pathway. (A and B) Both Wnt10a-siRNA and LGK-974 inhibitor decreased the expression of β-catenin, Akt, cyclin D1 and LEF1 in HCT116 cells. (C) Effect of Wnt10a-siRNA and LGK-974 on HCT116 cell growth at 48 h following treatment with Wnt10a-siRNA or LGK-974. Relative proliferation rate was normalized to the cells at 0 h timepoint. *P<0.05. Akt, protein kinase B; LEF1, lymphoid enhancer-binding factor 1; NC, negative control; siRNA, small interfering RNA; Wnt10a, Wnt family member 10A.
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