Regulation of TMEM100 expression by epigenetic modification, effects on proliferation and invasion of esophageal squamous carcinoma

Figure 1 Relationship between low TMEM100 expression in esophageal cancer and patient survival and the effect of 5-AZA on TMEM100 expression in esophageal squamous cell carcinoma lines. A: Expression profile of TMEM100 in EC samples compared with normal samples, showing reduced expression of TMEM100 in EC tissues; B: Overall survival of patients with high vs low TMEM100 expression levels. Survival was poorer for those with low TMEM100 expression (P = 0.041); C: 5-AZA induced a dose-dependent expression of TMEM100 in K-150 cells. Real-time PCR and western blotting results showed that after 24 h of treatment, TMEM100 expression increased with increasing 5-AZA concentration. ^aP < 0.05, ^bP < 0.01. DMSO: Dimethyl sulfoxide; EC: Esophageal cancer; ESCA: Esophageal cancer; HR: Hazard Ratio.

Figure 2 Overexpression effect of TMEM100 in esophageal squamous cell carcinoma lines and the inhibitory effect of TMEM100 overexpression on proliferation, migration, and invasion of esophageal squamous cell carcinoma cells in vitro. A and B: K-150/K-450 cells transfected with TMEM100-oe were assayed using real-time PCR and western blotting, and the results showed that the expression of TMEM100 was significantly upregulated in the transfected cells compared to that in the control group; C: Colony formation viability of K-150/K-450 cells after transient transfection treatment for 14 d was analysed by staining with 1% crystal violet; D: Cell counting kit-8 assay results show that overexpression of TMEM100 inhibits the proliferation of K-150/K-450 cells. ^cP < 0.0001.

Figure 3 Identification of differentially expressed genes and functional enrichment analysis. A: In the volcano plot, upregulated genes are indicated by red dots, and downregulated genes are indicated by green dots; B: The heatmap represents the expression levels of the genes, with the blue to red spectrum indicating low to high expression; C and D: The top 69 enriched Kyoto Encyclopedia of Genes and Genomes pathways.

Figure 4 Effect of TMEM100 overexpression on mitogen-activated protein kinase pathway activation in KYSE-150/KYSE-450 cells. A: K-150 cells were harvested 24 h after transfection with TMEM100-oe, and total proteins were extracted for western blotting analysis. Phosphorylated-extracellular regulated kinase (p-ERK) and ERK, phosphorylated-c-Jun N-terminal kinase (p-JNK) and JNK, and p-p38 and p38 were analysed. The result demonstrated a reduction in the expression of p-ERK, p-p38, and p-JNK in K-150/K-450 cells transfected with TMEM100-oe; B: The experiment was repeated again with K-450 cells. p-ERK: Phosphorylated-extracellular regulated kinase; p-JNK: Phosphorylated-c-Jun N-terminal kinase.