Effect of IFN-γ on the expression of IL-33 in colon cancer cells via PKA-CREB pathway / 临床检验杂志

ABSTRACT

Objective@#To investigate the effect of IFN-γ on the expression of IL-33 in colon cancer CT26 cells. @*Methods@#CT26 cells were treated with IFN-γ and IFN-γ combined with PKA inhibitor H89, respectively, and a negative control group (NC, untreated) was set up at the same time. The mRNA expression levels of PKA, CREB and IL-33 in CT26 cells were detected by qRT-PCR. The expression levels of PKA, CREB, p-CREB and IL-33 proteins in CT26 cells were determined by western blot. The localization of CREB protein in CT26 cells was analyzed by the immunofluorescence confocal microscopy. @*Results@#The relative expression levels of PKA, CREB and IL-33 mRNA in the IFN-γ-treated group were 2.50±0.11, 3.10±0.08 and 2.80±0.22, respectively, which were significantly higher than those in the NC group (P＜0.05). The relative expression levels of PKA, CREB and IL-33 mRNA in the IFN-γ combined with H89 treatment group were 0.21±0.02, 0.59±0.05 and 0.35±0.04, respectively, which were significantly lower than those in the NC group and IFN-γ-treated group (P＜0.05). The expression levels of PKA, CREB and IL-33 proteins detected by western blot were consistent with that of mRNA. Immunofluorescence confocal results showed that the expression level of CREB in the IFN-γ-treated group was significantly higher than that in the NC group, and that the expression level of CREB in the IFN-γ combined with H89 treatment group was significantly lower than that in the IFN-γ-treated group. @*Conclusion@#IFN-γ may induce the expression of IL-33 in colon cancer CT26 cells via the PKA-CREB pathway.