Panoply™ Human IL21 Knockdown Stable Cell Line

Previous studies have shown elevated expression levels of interleukin-21 (IL-21) protein in clinical samples from colorectal cancer patients in Sudan, regardless of whether they had concurrent parasitic infections. In the present study, researchers further investigated the relationship between the IL-21 gene and processes such as cell proliferation, migration, and associated gene expression mechanisms in HT29 and HCT116 cells. The results demonstrated that knocking down the IL-21 gene in HCT116 cells increased cytotoxicity levels and the mRNA expression of fibroblast growth factor-4 (FGF4). Furthermore, specific knockdown of the IL-21 gene inhibited the migratory capacity of the cancer cells compared to cells without the knockdown; however, these effects were not observed in IL-21-knockdown HT29 cells. Compared to IL-21 knockdown alone, neutralizing FGF4 in the conditioned medium of IL-21-knockdown HCT116 cells further enhanced cytotoxicity and restored the migratory activity of the HCT116 cells. These findings highlight the significance of the interplay between IL-21 gene knockdown and FGF4 protein expression in HCT116 cells, laying the groundwork for identifying key biomarkers and facilitating the future development of drugs or cost-effective supplements to effectively treat patients suffering from both infectious diseases and colorectal cancer (specifically of the HCT116 cell type).

Experimental results showed that, compared to control HCT116 cells, IL-21-knockdown HCT116 cells failed to completely close the wound-which initially spanned the full width of the scratch (100%)-by the end of the experiment. At 72 hours post-initiation, the residual wound rates for control and IL-21-knockdown HCT116 cells were 38.8% and 76.9%, respectively (Figure 1a). However, when these knockdown colorectal cancer cells were cultured in medium containing an FGF4-neutralizing antibody, the wounds in the IL-21-knockdown HCT116 cells closed, resulting in a residual rate of only 44.2% after 72 hours. In summary, IL-21-knockdown HCT116 cells exhibited a relatively slower invasion rate compared to control HCT116 cells (Figure 1b); real-time PCR analysis confirmed the presence of FGF4 expression in the IL-21-knockdown HCT116 cells. Notably, culturing the IL-21-knockdown HCT116 cells in medium containing the FGF4-neutralizing antibody restored their activity, indicating that neutralizing FGF4 in the conditioned medium rescued the migratory activity of the IL-21-knockdown cells. Therefore, the combination of reduced IL-21 expression and FGF4 expression is essential for the inhibition of HCT116 cell migration.

Figure 1. Migration of IL-21-knockdown HCT116 cells. (Abdalkareem E A, et al., 2018)