Fig. 8
Impact of AKR1C3 Regulation on NF-κB Localization and Function in GC Cells. (a) Overexpression of AKR1C3 decreased phosphorylation of IκBα and IKKα/β, whereas AKR1C3 knockdown led to increased phosphorylation of these proteins. (b) The increase in IKKα/β phosphorylation levels in response to shAKR1C3 can be inhibited by SP600125 (a JNK inhibitor). (c) Cell fractionation experiments showed that upregulation of AKR1C3 led to decreased expression of p-NF-kB in both the nucleus and cytosol of AGS and SNU-216 cells, while downregulation of AKR1C3 enhanced p-NF-kB expression. GAPDH was utilized as a loading control, with LaminB and GAPDH serving as markers for nuclear and cytosolic fractions, respectively. *P < 0.05, unpaired t test, pCDH versus pAKR1C3. *P < 0.05, one-way ANOVA with Dunnett’s post hoc test, pSuper versus psh1AKR1C3 and pSuper versus psh2AKR1C3. (d) Schematic model illustrating how AKR1C3 knockdown promotes malignant phenotypes in GC by activating p-JNK, which in turn activates phosphorylation of IKKα/β, leading to increased phosphorylation and activation of IκBα. This process facilitates the release and nuclear entry of p-NF-κB, which promotes EMT by upregulating transcription factors Snail and Slug, downregulating epithelial markers (E-cadherin and ZO-1), and upregulating mesenchymal markers (vimentin and N-cadherin).
