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. 2021 Sep 17;59(5):86. doi: 10.3892/ijo.2021.5266

Figure 6.

Figure 6

SPHK1 is a direct target of miR-378a-5p in ESCC cells. (A) TargetScan and miRDB both predicted SPHK1 as the downstream target of miR-378a-5p. (B) A dual-luciferase reporter assay was performed to investigate the binding status of SPHK1 and miR-378a-5p in KYSE150 and KYSE30 cells. (C) Ago2-RIP analysis of the enrichment of SPHK1 in RNA-induced silencing complex in KYSE150 and KYSE30 cells. (D) Ago2-RIP analysis of the enrichment of SPHK1 in KYSE150 and KYSE30 cells after transfection with miR-378a-5p mimic. (E) Spearman's rank correlation analysis was used to determine the correlation between miR-378a-5p and SPHK1 expression in 85 ESCC and paired normal samples. (F) miR-378a-5p mimic significantly upregulated the expression of miR-378a-5p in KYSE150 and KYSE30 cells. (G) miR-378a-5p inhibitor markedly downregulated the expression of miR-378a-5p in KYSE150 and KYSE30 cells. (H) miR-378a-5p mimic suppressed the expression of SPHK1 in KYSE150 and KYSE30 cells. (I) miR-378a-5p inhibitor promoted the expression of SPHK1 in KYSE150 and KYSE30 cells. ***P<0.001, ****P<0.0001. SPHK1, sphingosine kinase 1; ESCC, esophageal squamous cell carcinoma; miR, microRNA; Ago2, argonaute 2; RIP, RNA immunoprecipitation.