Epigenetic silencing of JAM3 promotes esophageal cancer development by activating Wnt signaling

Abstract

Background The role of JAM3 in different tumors is controversial. The epigenetic regulation and the mechanism of JAM3 remain to be elucidated in human esophageal cancer (EC). Methods Eleven EC cell lines, 49 cases of esophageal intraepithelial neoplasia (EIN) and 760 cases of primary EC samples were employed. Methylation-specific polymerase chain reaction, immunohistochemistry, MTT, western blot and xenograft mouse models were applied in this study. Results The inverse association between RNA expression and promoter region methylation of JAM3 was found by analyzing 185 cases of EC samples extracted from the TCGA database (p < 0.05). JAM3 was highly expressed in KYSE450, KYSE520, TE1 and YES2 cells, low level expressed in KYSE70 cells and unexpressed in KYSE30, KYSE150, KYSE410, KYSE510, TE13 and BIC1 cells. JAM3 was unmethylated in KYSE450, KYSE520, TE1 and YES2 cells, partial methylated in KYSE70 cells and completely methylated in KYSE30, KYSE150, KYSE410, KYSE510, TE13 and BIC1 cells. The expression of JAM3 is correlated with methylation status. The levels of JAM3 were unchanged in KYSE450, KYSE520, TE1 and YES2 cells, increased in KYSE70 cells and restored expression in KYSE30, KYSE150, KYSE410, KYSE510, TE13 and BIC1 cells after 5-aza-2′-deoxycytidine treatment, suggesting that the expression of JAM3 is regulated by promoter region methylation. JAM3 was methylated in 26.5% (13/49) of EIN and 51.1% (388/760) of primary EC, and methylation of JAM3 was associated significantly with tumor differentiation and family history (all p < 0.05). Methylation of JAM3 is an independent prognostic factor of poor 5-year overall survival (p < 0.05). JAM3 suppresses cell proliferation, colony formation, migration and invasion and induces G1/S arrest and apoptosis in EC. Further study demonstrated that JAM3 suppressed EC cells and xenograft tumor growth by inhibiting Wnt/β-catenin signaling. Conclusion JAM3 is frequently methylated in human EC, and the expression of JAM3 is regulated by promoter region methylation. JAM3 methylation is an early detection and prognostic marker of EC. JAM3 suppresses EC growth both in vitro and in vivo by inhibiting Wnt signaling.

Identification of a tissue resident memory CD8 T cell-related risk score signature for colorectal cancer, the association with TME landscapes and therapeutic responses

“…By contrast, Trm cluster B was characterized by enhanced tumor purity and a paucity of immune and stromal contents in TME. Notably, Wnt_β-catenin pathway, a canonical carcinogenic pathway for malignancies in digestive system ( Xue et al, 2021 ; Wen et al, 2022 ; Yang et al, 2022 ; Zhang et al, 2022 ; Zhu et al, 2022 ), was the Hallmark item with the highest enrichment level in Trm cluster B ( Figure 3B ), which reflected the high tumor purity of this cluster.…”

Downregulation of JAM3 occurs in cholangiocarcinoma by hypermethylation: A potential molecular marker for diagnosis and prognosis

“…The methylation status of the CpG island in the region related to the JAM3 promoter (Figure 2A) in RBE, HCCC‐9810 and HuCCT1 cells was checked by bisulfite‐treated DNA direct sequencing (BSP) 23 . The hypermethylated CpG island in the region related to the promoter of JAM3 was detected in HCCC‐9810 and HuCCT1 cells but not in RBE cells (Figure 2B,C,D).…”

“…Leukaemia‐initiating cells, which present high JAM3 expression, are important for the initiation, development, and relapse of leukaemia, while knockdown of JAM3 has been shown to dramatically decrease the proliferation of leukaemia cell lines and primary leukaemia‐initiating cells 25 . Junctional adhesion molecular 3 has been shown to be downregulated by hypermethylation in cervical cancer, colorectal cancer, and oesophageal cancer 17,22,23 . Moreover, JAM3 methylation status in cervical tissues and scraping demonstrated a significant difference between low‐ and high‐grade cervical intraepithelial neoplasia (CIN) and normal cervical tissue and CIN, suggesting that JAM3 methylation could be a biomarker for the diagnosis of cervical cancer and for screening 22 .…”

“…Junctional adhesion molecular 3 also suppresses cell proliferation, colony formation, migration and invasion and induces cell cycle arrest and apoptosis via Wnt/β‐catenin signalling. The methylation of JAM3 is an independent prognostic factor of overall survival for oesophageal cancer 23 …”

“…The methylation of JAM3 is an independent prognostic factor of overall survival for oesophageal cancer. 23 Although the expression of JAM3 has been reported previously in different human cancers, its biological role remains controversial. To the best of our knowledge, the expression profile, methylation status and biological role of JAM3 in CCA are unknown.…”