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  • br Conclusions Our study revealed that high expression of GPR


    Conclusions: Our study revealed that high 2-NBDG of GPR110 predicts the poor prognosis of GC patients, and GPTR110 may function as a potential biomarker for the diagnosis of GC.
    1. Introduction
    Gastric cancer (GC) is an etiologically multifactorial disease which represents one of the most common malignant tumors worldwide [1]. Although, the therapeutic tools of GC are varied including surgery, chemotherapy, radiotherapy, gene therapy and other options, the morbidity and mortality rates still remain high globally, especially in China [2,3]. Therefore, there is an urgent need to explore new diag-nostic and prognostic targets for GC patients.
    Adhesion GPCRs are characterized by extremely long N-terminal regions and a GPCR-like-seven-pass transmembrane domain [4]. Recent years, several adhesion GPCR molecules have been identified associated with cancer development in various cancers [5–9]. GPR110, a number of the adhesion G protein-coupled receptor family, has also been identified as an oncogene during the development of several cancers, including lung and prostate adenocarcinomas, breast cancer and he-patocellular carcinoma [10–13]. In 2010, Amy et al. identified that orphan receptor GPR110 was overexpressed in lung and prostate cancer
    Corresponding author.
    [10]. In addition, ma et al. revealed that GPR110 deficiency decelerates carcinogen-induced hepatocarcinogenesis via activation of the IL-6/ STAT3 pathway [11]. Furthermore, a study published recently revealed that GPR110 could function as a potential new target in HER2+ breast cancer through GPCR profiling [12]. However, to the best of our knowledge, the expression of GPR110, as well as its prognostic sig-nificance, has not been investigated in GC.
    In the present study, the mRNA and protein levels of GPR110 were examined in 117 paired GC specimens and adjacent non-tumorous tis-sues. The association between GPR110 expression and the clin-icopathological parameters as well as the 5 year overall and recurrence-free survival of GC patients was analyzed. The results indicate that GPR110 may be a predictive biomarker for the poor prognosis in pa-tients with GC.
    Table 1
    Relationship between GPR110 expression in tumorous tissues and clin-icopathological characteristics in patients with gastric cancer.
    Clinicopathological N GPR110
    Pearson χ2 P-value
    Note: Bold values have statistical significance.
    2. Material and methods
    2.1. Patients and samples
    We obtained 117 paired GC tissues and adjacent non-tumorous tissues from patients who underwent a radical resection from the same surgical team and were histologically and clinically diagnosed with GC. None of the patients had received preoperative treatments, for example chemotherapy, radiotherapy or other related anti-tumor treatments. Surgeries were performed at the Department of Gastrointestinal Surgery, the Affiliated Wenling Hospital of Wenzhou Medical University between June 2010 and November 2012. The clin-icopathological parameters, including age, gender, tumor differentia-tion, tumor size, lymph node metastasis, tumor stage and TNM stage were retrospectively collected. All tissue samples were immediately collected and cryopreserved in liquid nitrogen for further study. A portion of each specimen was fixed with 10% paraformaldehyde and embedded in paraffin blocks. Written informed consent was obtained from the patients prior to participation in this clinical trial and re-search, and the research protocols were approved by the ethics com-mittee of the Affiliated Wenling Hospital of Wenzhou Medical University. The clinicopathological characteristics of the patients are presented in Table 1.
    2.2. RNA isolation and quantitative real-time PCR (qRT-PCR)
    Total RNA was extracted from tissue sample using TRIzol reagent (Invitrogen, Grand Island, NY, USA) as described by the manufacturer and RNAs (500 ng) were reverse transcribed using the PrimeScript RT Master Mix (Takara, Dalian, China). Quantitative real-time PCR was performed to detect the expression levels of GPR110 using the SYBR Premix Ex Taq (Takara, Dalian, China) on the ABI Prism 7900 H T (Applied Biosystems, Foster City, CA, USA). The relative expression levels of GPR110 were normalized to GAPDH. The reactions were in-cubated in a 384-well optical plate at 95 °C for 30 s, followed by 40 cycles of 95 °C for 5 s and 60 °C for 30 s. The primer sequences were as