Panoply™ Human MAPK14 Knockdown Stable Cell Line

Mitogen-activated protein kinase 14 (MAPK14) plays a crucial role in DNA damage and repair and can be activated by various environmental stresses and pro-inflammatory cytokines. It is highly active in many tumors, acting as both a tumor promoter and a tumor suppressor, but its role in clear cell renal cell carcinoma (ccRCC) remains unclear. Cell cycle phosphatase 25B (CDC25B) is involved in cell cycle regulation and is highly expressed in many malignant tumors. Here, researchers analyzed the transcriptional levels of MAPK14 and CDC25B in 72 pairs of ccRCC and adjacent normal tissue samples from The Cancer Genome Atlas database, as well as the protein expression levels of MAPK14 and CDC25B in 66 pairs of clinical samples. Knockdown of MAPK14 reduced the protein levels of phosphorylated MAPK14 (P-MAPK14) and CDC25B. Subsequent Western blot and co-immunoprecipitation experiments showed that P-MAPK14 can bind to CDC25B, potentially maintaining its stability. Knockdown of MAPK14 inhibited the proliferation and migration of ccRCC cell lines, but this inhibition was partially reversed by overexpression of CDC25B. These results suggest that downregulation of MAPK14 and P-MAPK14 may inhibit ccRCC proliferation and migration by downregulating CDC25B.

To further investigate the biological function of MAPK14 in clear cell renal cell carcinoma (ccRCC), researchers used the CCK-8 assay to detect the activity of MAPK14 knockdown cells. The results showed that the activity of MAPK14-knockdown ACHN and CAKI-1 cells was significantly reduced compared to the control group (Figure 1A). EdU assay results indicated that cell proliferation capacity was significantly decreased after MAPK14 knockdown (Figure 1B). These results suggest that knocking down MAPK14 can inhibit the activity and proliferation of ccRCC cells in vitro. Furthermore, the migration ability of MAPK14-knockdown ACHN and CAKI-1 cells was significantly reduced (Figure 1C). Since the upregulation of E-cadherin and downregulation of N-cadherin have been confirmed to be associated with decreased cell migration ability, the researchers used Western blot to detect the expression of E-cadherin and N-cadherin after MAPK14 knockdown to further verify this observation. In MAPK14-knockdown cells, the expression of E-cadherin was significantly upregulated compared to the control group, while the expression of N-cadherin was significantly downregulated (Figure 1D). These results suggest that MAPK14 may play an important role in cell activity, proliferation, and migration.

Figure 1. Downregulation of MAPK14 inhibited cell activity, proliferation, and migration. (Liu J, et al., 2020)