Knocking down GALNT6 promotes pyroptosis of pancreatic ductal adenocarcinoma cells through NF-κB/NLRP3/GSDMD and GSDME signaling pathway

Abstract

BackgroundPancreatic ductal adenocarcinoma (PDAC), the most prevalent type of pancreatic cancer, is a highly lethal malignancy with poor prognosis. Polypeptide N-acetylgalactosaminyltransferase-6 (GALNT6) is frequently overexpressed in PDAC. However, the role of GALNT6 in the PDAC remains unclear.MethodsThe expression of GALNT6 in pancreatic cancer and normal tissues were analyzed by bioinformatic analyses and immunohistochemistry. CCK8 and colony formation were used to detect cell proliferation. Flow cytometry was applied to detect cell cycle.The pyroptosis was detected by scanning electron microscopy. The mRNA expression was detected by qRT-PCR. The protein expression and localization were detected by western blot and immunofluorescence assay. ELISA was used to detect the levels of inflammatory factors.ResultsThe expression of GALNT6 was associated with advanced tumor stage, and had an area under curve (AUC) value of 0.919 in pancreatic cancer based on the cancer genome atlas (TCGA) dataset. Knockdown of GALNT6 inhibited cell proliferation, migration, invasion and cell cycle arrest of PDAC cells. Meanwhile, knockdown of GALNT6 increased the expression levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-18 (IL-18), the release of inflammasome and an increasing of Gasdermin D (GSDMD), N-terminal of GSDMD (GSDMD-N), Gasdermin E (GSDME) and N-terminal of GSDME (GSDME-N) in PDAC cells. GALNT6 suppressed the expression of NOD-like receptor thermal protein domain associated protein 3 (NLRP3) and GSDMD by glycosylation of NF-κB and inhibiting the nucleus localization of NF-κB. Additionally, GALNT6 promotes the degradation of GSDME by O-glycosylation.ConclusionWe found that GALNT6 is highly expressed in pancreatic cancer and plays a carcinogenic role. The results suggested that GALNT6 regulates the pyroptosis of PDAC cells through NF-κB/NLRP3/GSDMD and GSDME signaling. Our study might provides novel insights into the roles of GALNT6 in PDAC progression.

KIF23 promotes cervical cancer progression via inhibiting NLRP3‐mediated pyroptosis

“…37 For example, GALNT6 serves as a carcinogenic role in pancreatic ductal adenocarcinoma cells through regulation pyroptosis, which is mediated via NF-κB/NLRP3/GSDMD and GSDME signaling. 38 FABP4 is involved in inducing macrophage pyroptosis via NLRP3/IL-1β axis, thereby promoting the migration, invasion, and metastasis in pancreatic cancer cells. 39 Hydrogen inhibits endometrial cancer progression, which is linked to pyroptotic pathway via ROS/ NLRP3/caspase-1/GSDMD.…”

“…Increasing evidence has demonstrated that pyroptosis contributes to the onset and progression of cancers, in which the NLRP3 inflammasome‐mediated pathway is particularly highlighted 37 . For example, GALNT6 serves as a carcinogenic role in pancreatic ductal adenocarcinoma cells through regulation pyroptosis, which is mediated via NF‐κB/NLRP3/GSDMD and GSDME signaling 38 . FABP4 is involved in inducing macrophage pyroptosis via NLRP3/IL‐1β axis, thereby promoting the migration, invasion, and metastasis in pancreatic cancer cells 39 .…”

Low expression of GALNT6 gene in gliomas promotes immune infiltration and improves tumor prognosis using bioinformatics analysis

“…Up until now, GALNT6 has been discovered to be extremely conserved in the human genome, playing a role in both cell proliferation and differentiation, as well as contributing to the development of tumors. We reviewed the literature and found that GALNT6 is aberrantly expressed in many tumour cells, such as breast invasive carcinoma (BRCA) 34 , Ovarian serous cystadenocarcinoma (OV) 35 , pancreatic ductal adenocarcinoma 36 , gastric cancer 37 , Lung adenocarcinoma (LUAD) 38 , colon adenocarcinoma 39 Various types of lymphocytes have been connected to the dissemination and metastasis of tumors, and they have also been correlated with the immune Studies have demonstrated that various elements released by macrophages play a role in enhancing the growth and/or movement of glioma cells. In vitro and in vivo, stress-responsive protein 1 (STI1), an intracellular PrP protein ligand, synthesized and discharged by macrophages, stimulates the proliferation and migration of glioma.…”

FUT2 promotes colorectal cancer metastasis by reprogramming fatty acid metabolism via YAP/TAZ signaling and SREBP-1

“…Western blotting was performed as previously described [50]. The primary antibodies were used in Tab.S3.…”

“…Quantitative real-time PCR was performed as previously described [50]. The primer sequences were used in Tab.S2.…”