Over expression of GPR30, indicating poor prognosis and promoting proliferation, upregulates Beclin-1 expression via p38MAPK signaling in esophageal squamous cell carcinoma progression.

BACKGROUND: Beclin-1 and GPR30, both very important proteins, have been associated with tumor development. In our pre-experiment, the co-expression of GPR30 and Beclin-1 was observed in esophageal squamous cell carcinoma (ESCC), an observation not reported in other studies. The aim of our research was to investigate the relationship of these two proteins in the further progression of ESCC.
METHODS: The over expression of GPR30 and Beclin-1 proteins was observed and confirmed by immunohistochemistry and immunofluorescence arrays. By interfering with GPR30 and p38 MAPK expression in EC-109, KYSE510, and KYSE3 cell lines, MTT and a scratch wound healing assay were used to investigate the impact of the GPR30 protein on the proliferative and migrative abilities of ESCC cells. A co-immunoprecipitation assay was used to observe the interaction between the p38 MAPK and Beclin-1 proteins; meanwhile, at a different time, in each group, the GPR30, MAPK, p ERK1/2, p38 MAPK, and Beclin-1 proteins were analyzed. The correlation between GPR30, Beclin-1 expression levels, and the clinical characteristics were evaluated by Mann-Whitney and chi-square tests. Using Kaplan-Meier plots and the Cox proportional hazard model analysis, we determined overall survival (OS) and progression free survival (PFS).
RESULTS: GPR30 and Beclin-1 were over expressed significantly in ESCC (both p=0.0000) and were distributed into cytoplasms the most (the former p=0.0223, latter p=0.0018). In contrast to the non-agonist group, the abilities of GPR30 in promoting cell proliferation and metastasis were observed in the agonist group, and the effects could be blocked by p38MAPK inhibitors. In further assays, GPR30 agonists, via binding to GPR30, up-regulated Beclin-1, MAPK, and p38 MAPK expression, and Beclin-1 expression was reversed in the p38MAPK inhibitor group. In the GPR30 agonist group, an interaction between p38MARK and Beclin-1 was observed, but no similar results were observed in the non-agonist group. The high expression of both GPR30 and Beclin-1 was observed with p-stage and pT advancing (both p<0.0001). In a Kaplan-Meier analysis, compared to GPR30's negative expression, high expression identified a group of patients with the shortest overall survival (OS, p=0.0072) and progression free survival (PFS, p=0.0074). The Cox proportional hazard models revealed that they predicted a short time to OS (p=0.0125).
CONCLUSION: Over expression of GPR30 up-regulated Beclin-1 expression and indicated a poor prognosis and may promote ESCC development via p38 MAPK in ESCC progression. IJCEP