Panoply™ Human WWP1 Over-expressing Stable Cell Line

Protein tyrosine phosphatase SHP2 activates the RAS signaling pathway, a novel target for colorectal cancer (CRC) treatment. However, SHP2 inhibitor monotherapy is ineffective against metastatic CRC, necessitating combination therapy. Here, researchers demonstrate that WWP1 promotes CRC cell proliferation. Genetic or pharmacological inhibition of WWP1 enhances the tumor-inhibiting effects of SHP2 inhibitors both in vitro and in vivo. WWP1 may mediate feedback reactivation of the AKT signaling pathway following SHP2 inhibition. Furthermore, a nomogram model based on the immunohistochemical expression of WWP1 and SHP2 significantly improved the accuracy of prognostic prediction for CRC patients. These results suggest that the WWP1 inhibitor I3C can synergize with SHP2 inhibitors, potentially representing a novel clinical trial strategy for treating advanced CRC patients.

Here, researchers examined the expression levels of WWP1 in several CRC cell lines with different mutation states (KRAS mutants: SW480, HCT116; BRAF mutants: RKO, HT29; wild-type: Caco-2, CW2) and normal colonic epithelial cell lines (FHC). The results showed that the expression level of endogenous WWP1 in CRC cells was higher than that in FHC cells (Figure 1a, b). Specifically, the WWP1 expression levels in CW2, HCT116, and HT29 cells were relatively low, while those in Caco-2, SW480, and RKO cells were relatively high (Figure 1a, b). The researchers then constructed WWP1-overexpressing cell lines in CW2, HCT116, and HT29 (Figure 1c, d). To avoid off-target effects, the researchers designed two different shRNAs targeting WWP1 and transfected Caco-2, SW480, and RKO cells. The knockdown efficiency of shRNA was verified by comparing the mRNA and protein levels of WWP1 in stably transfected cells and negative control cells (Fig. 1e, f). CCK-8 and colony formation assays showed that, compared to control cells, WWP1-overexpressing CW2, HCT116, and HT29 cells exhibited significantly enhanced cell proliferation and tumorigenicity (Fig. 1g, i). Conversely, WWP1 knockdown in Caco-2, SW480, and RKO cells significantly reduced cell proliferation and tumorigenicity (Fig. 1h, j). These results reveal the crucial role of WWP1 in promoting the in vitro proliferation of CRC cells.

Figure 1. WWP1 promotes CRC cells proliferation in vitro. (Fan H, et al., 2024)