Sulforaphene inhibits esophageal cancer progression via suppressing SCD and CDH3 expression, and activating the GADD45B-MAP2K3-p38-p53 feedback loop

Abstract

Esophageal cancer is one of the most common cancer with limited therapeutic strategies, thus it is important to develop more effective strategies to against it. Sulforaphene (SFE), an isothiocyanate isolated from radish seeds, was proved to inhibit esophageal cancer progression in the current study. Flow cytometric analysis showed SFE induced cell apoptosis and cycle arrest in G2/M phase. Also, scrape motility and transwell assays presented SFE reduced esophageal cancer cell metastasis. Microarray results showed the influence of SFE on esophageal cancer cells was related with stearoyl-CoA desaturase (SCD), cadherin 3 (CDH3), mitogen-activated protein kinase kinase 3 (MAP2K3) and growth arrest and DNA damage inducible beta (GADD45B). SCD and CDH3 could promote esophageal cancer metastasis via activating the Wnt pathway, while the latter one was involved in a positive feedback loop, GADD45B-MAP2K3-p38-p53, to suppress esophageal cancer growth. GADD45B was known to be the target gene of p53, and we proved in this study, it could increase the phosphorylation level of MAP2K3 in esophageal cancer cells, activating p38 and p53 in turn. SFE treatment elevated MAP2K3 and GADD45B expression and further stimulated this feedback loop to better exert antitumor effect. In summary, these results demonstrated that SFE had the potential for developing as a chemotherapeutic agent because of its inhibitory effects on esophageal cancer metastasis and proliferation.

Identifying the p65-Dependent Effect of Sulforaphene on Esophageal Squamous Cell Carcinoma Progression via Bioinformatics Analysis

“…We next sought to determine whether TNFAIP3, PLAU, and their regulator p65 were involved in SFE-mediated inhibition of ESCC cell progression. We previously confirmed that SFE induced ESCC cell G2/M arrest and apoptosis to inhibit cell proliferation, and reduced the invasive and migratory abilities of ESCC cells [9]. Flow cytometry analyses, scrape motility, and trans-well assays shown in Figure 5 also suggested SFE inhibition of ESCC cell progression.…”

“…All these genes were overexpressed in the TCGA ESCC samples compared with in the normal tissues collected in the TCGA database or the TCGA and the GTEx databases (Supplementary Figure S3, Figure 2C). We then performed qRT-PCR in SFE-treated ESCC cells (Figure 3A) and ground tumor lumps (Figure 3B) from the previous xenograft tumor assay [9], verifying that the expression of CXCL10, TNFAIP3, INHBA, and PLAU was indeed controlled by SFE in ESCC.…”

“…We have verified SCD, CDH3, MAP2K3 and GADD45B as the targets of SFE in ESCC through microarray analyses [9]; however, changing the expression of these genes could not completely reverse the inhibitory effect of SFE on ESCC proliferation and metastasis, which suggests that there must be some other mediators to connect SFE with ESCC characteristics. Since confirming the mechanism by which SFE inhibits ESCC progression contributes to the application of this isothiocyanate as a chemotherapeutic agent, we attempted to investigate SFE regulation of ESCC progression more deeply.…”

“…To shed more light on SFE-induced inhibition of ESCC tumorigenesis and development, we conducted GSEA to reanalyze previously described microarray results (GSE150891) [9] (Supplementary Figure S1A) and set the cutoff criteria as p < 0.05 and false discovery rate (FDR) < 0.05. We found that the significantly differentially expressed genes (DEGs) in ESCC cells treated with dimethyl sulfoxide (DMSO) as negative control were mainly enriched in pathways related to immunity, cell proliferation, and metastasis, such as epithelial mesenchymal transition, up-regulated KRAS proto-oncogene, GTPase (KRAS) signaling; IL6, Jak, and STAT3 signaling and IL2 and STAT5 signaling (Supplementary Figure S1B).…”

“…It inhibited triple-negative breast cancer by activating the tumor suppressor Egr1 [6], induced hepatocellular carcinoma cell death by repressing keratin 8 and promoting anoikis [7], and targeted the PI3K-Akt pathway to cause lung cancer cell apoptosis [8]. We previously emphasized that SFE could also block ESCC progression by suppressing stearoyl-CoA desaturase (SCD) and cadherin 3 (CDH3) expression and boosting the GADD45B-MAP2K3-p38-p53 positive feedback loop [9].…”