Fig. 6: The OGT-POM121-c-MYC-ECM regulatory axis is conserved in H1299 NSCLC cell model. | Oncogene

Fig. 6: The OGT-POM121-c-MYC-ECM regulatory axis is conserved in H1299 NSCLC cell model.

From: POM121 O-GlcNAcylation facilitates bone metastasis in non-small cell lung cancer through enhanced c-MYC nuclear import and ECM reprogramming

Fig. 6

A Analysis of POM121 and OGT expression between parental H1299 and bone-metastatic H1299-BM sublines. Data representative of three experiments. B, C O-GlcNAcylation stabilizes POM121 in H1299-BM metastatic cells. B sWGA pulldown of O-GlcNAcylated proteins from H1299 and H1299-BM lysates, immunoblotted for POM121. C Analysis POM121 stability in H1299-BM transfected with OGT siRNAs and H1299 transfected with 3×Flag-OGT plasmid. Data representative of three experiments. D O-GlcNAcylated POM121 exhibited significantly attenuated binding affinity to TRIM21 in metastatic lineage. H1299 and H1299-BM cells treated with MG132 (25 nM, 24 h). Cell lysates IP with anti-POM121 and immunoblotted for TRIM21 and ubiquitin. Data representative of three experiments. E Enhanced nuclear c-MYC accumulation in metastatic cells. Immunoblots of c-MYC levels in the nucleoplasm and cytoplasm of H1299 cells and H1299-BM cells. HSP90 and UAP56 served as fractionation controls. Data representative of three experiments. F Left, ECM-related genes mRNA levels were analyzed by RT-qPCR between H1299 and H1299-BM cells. (mean ± SEM; n = 3; unpaired t-test). Right, Analysis of PI3K-AKT-mTOR pathway activation (p-PI3K/PI3K, p-AKT/AKT, p-mTOR/mTOR) between H1299 and H1299-BM cells. Data representative of three experiments. G Pan-cancer analysis showed that POM121 was highly expressed in NSCLC. (TCGA data; *p < 0.05, **p < 0.01, ***p < 0.001). H High POM121 and c-MYC co-expression correlates with poor NSCLC prognosis. Kaplan-Meier overall survival analysis of 2166 NSCLC patients from TCGA, stratified by high/low mRNA expression of POM121 (log-rank test, p < 0.05). I Significantly elevated POM121 and c-MYC expression in bone metastatic lesions versus primary lung lesions of LUAD patients with bone metastasis. Data derived from the GSE225208 dataset (bulk RNA-seq; n = 20 primary lung lesions vs. n = 7 bone metastatic lesions, unpaired t-test, ***p < 0.001, ****p < 0.0001).

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