Then, an RNase protection assay was used to verify the presence of the overlapping and non‐overlapping regions in expressed mRNA using RT‐qPCR; the data indicated that the overlapping region was partially protected from RNase degradation in ESCC cells with SLC25A21‐AS1 overexpression (Figure 6E). This evidence concerns the gene SLC25A21 and esophageal squamous cell carcinoma.