LRFN2 binding to NMDAR inhibits the progress of ESCC via regulating the Wnt/ß-Catenin and NF-κB signaling pathway.

As a neuronal transmembrane protein, leucine-rich repeat and fibronectin type-III domain-containing protein 2 (LRFN2) can recruit and combine with N-methyl-d-aspartate receptors (NMDARs) to promote nerve growth. Genetic studies suggest that mutations in LRFN2 are associated with various cancers. However, the role and mechanism of LRFN2 in the progression of ESCC have not been elucidated. In this study, we demonstrated that LRFN2 was significantly downregulated in ESCC tissues by qRT-PCR and immunohistochemistry. Low LRFN2 expression was an adverse prognostic factor in patients with ESCC. Overexpression of LRFN2 effectively suppressed the proliferation, migration, invasion, and epithelial-to-mesenchymal transition in vitro and tumor growth in vivo. Bioinformatics analysis indicated that Wnt/ß-catenin signaling regulation was one of the most potential mechanisms and studies confirmed that overexpression of LFRN2 obviously downregulated the expression of ß-catenin, c-Myc, and cyclin D1 in ESCC cells and tumor tissues. Further studies revealed that LRFN2 plays an anti-ESCC role by binding with NMDAR-GRIN2B and this effect can be weakened by NR2B-selective NMDA antagonist-NMDA-IN-1. Moreover, the bioinformatics analysis showed that the interaction of GRIN2B and GSK3ß affects the NF-κB pathway, which was demonstrated by western blot experiments. Collectively, our results indicate that LRFN2 binding to NMDARs inhibits the progression of ESCC by regulating the Wnt/ß-catenin and NF-κB pathway, which provides a new therapeutic target for improving the prognosis of patients with ESCC.

Nano-pearl powder/chitosan-hyaluronic acid porous composite scaffold and preliminary study of its osteogenesis mechanism.

Bone tissue engineering using osteoinductive scaffolds has evolved into a promising approach for bone regeneration. Pearl powder has recently gained interest in bone regeneration due to their bioactive characteristics and favorable mechanical properties. In order to mimic bone tissue structurally and compositionally, chitosan-hyaluronic acid (C-HA) scaffolds containing nano-pearl powder (NPP) were fabricated in current study by using freeze-drying method. The microstructure, porosity, hydrophilia, and mechanical property of scaffolds were studied by scanning electron microscopy (SEM), mercury porosimeter, contact angles, and universal material experiment machine respectively. The crystal form of NPP, which was packed in scaffolds, was tested by X-ray diffraction (XRD), and the interaction between three constituents was studied by Fourier transformed infrared (FTIR). In the other hand, the biocompatibility, osteogenic characteristics, and osteogenic related gene expression of scaffolds were examined by cell counting kit-8 (CCK-8), alkaline phosphatase activity (ALP), RT-qPCR, and Western blotting. The results revealed that the NPP was successfully incorporated in the C-HA scaffolds. The hydrophilia and mechanical properties of scaffolds increased with NPP content, whereas the pore morphology was disturbed, especially for 25% scaffold. The scaffolds had a high porosity between 89% and 93%, and decreased slightly with the increase of NPP. Cell culture tests indicated that scaffolds with higher NPP proportion were beneficial for the proliferation and differentiation of MC3T3-E1 cells, and scaffolds with 10 wt% and 25 wt% nano-pearl powder were most effective. The differentiation might be promoted by upregulating the expression of Col αI, OCN, OPN and Runx2 genes. Therefore, the nano-pearl powder/chitosan-hyaluronic acid (NPP/C-HA) scaffolds may serve as a promising biomaterial for bone tissue engineering.

PRAF2 overexpression predicts poor prognosis and promotes tumorigenesis in esophageal squamous cell carcinoma.

BACKGROUND: Prenylated Rab acceptor 1 domain family, member 2 (PRAF2) is involved in the occurrence and progression of several malignant tumors. However, its potential role in esophageal squamous cell carcinoma (ESCC) is still unknown. METHODS: PRAF2 mRNA expression was determined in 77 frozen ESCC samples by quantitative reverse transcription-polymerase chain reaction (qPCR) and its association with clinical features and overall survival were evaluated. The roles of PRAF2 in ESCC cells were investigated by proliferation, cell cycle, invasion and apoptosis assays in vitro. RESULTS: The PRAF2 mRNA expression was significantly increased in ESCC tissues compared with matched surrounding non-tumor tissues. Survival analysis showed that high PRAF2 mRNA expression was associated with worse overall survival in ESCC patients. Multivariate analysis revealed that PRAF2 (hazard ratio 2.05, 95% CI 1.10-3.85, P = 0.025) emerged as the independent predictor for poor overall survival in ESCC. The in vitro experiments revealed that knockdown of PRAF2 expression blocked cell proliferation, cell cycle progression and cell invasion and induced cell apoptosis in ESCC cells. CONCLUSION: Taken together, our data demonstrate that PRAF2 could be used as a potential prognostic biomarker and represent a potential therapeutic target for ESCC.

Intronic polymorphisms in genes LRFN2 (rs2494938) and DNAH11 (rs2285947) are prognostic indicators of esophageal squamous cell carcinoma.

BACKGROUND: Genome wide association study (GWAS) has become the major means to screen for the genetic variants associated with risk and prognosis of different diseases. A recent GWAS has discovered three novel intronic single nucleotide polymorphisms in genes LRFN2 (rs2494938), DNAH11 (rs2285947) and PLCXD2 (rs2399395) that are associated with altered risk of esophageal squamous cell carcinoma (ESCC) among Han Chinese populations. However, the prognostic significance of these variations in ESCC remains unclear. METHODS: To investigate the association of three novel single nucleotide polymorphisms (rs2494938, rs2285947, rs2399395) with the prognosis of ESCC patients, we recruited 287 ESCC patients treated with surgical resection and evaluated the potential significance of the three polymorphisms through Kaplan-Meier survival analysis, log-rank test, and Cox proportional hazards regression models. RESULTS: The ESCC patients carrying genotype AA at rs2494938 had worse survival and genotype GG at 2285947 had better prognosis (Log-rank P = 0.003 and Log-rank P = 0.037, respectively). In addition, rs2494938 at 6p21.1 was independently associated with overall survival of ESCC patients in recessive model [AA vs. GG/GA, HR = 3.12, 95% CI = 1.43-6.83, P = 0.004], rs2285947 at 7p15.3 was independently associated with overall survival of ESCC patients in both dominant model [AA/GA vs. GG, HR = 1.59, 95% CI = 1.02-2.49, P = 0.042] and additive model [AA vs. GA vs. GG, HR = 1.45, 95% CI = 1.05-2.01, P = 0.025]. CONCLUSIONS: This study demonstrated that the polymorphisms rs2494938 at 6p21.1 and rs2285947 at 7p15.3 may serve as independent prognostic biomarkers for ESCC, implying the potential biological role of their related genes (LRFN2 and DNAH11) in the process of ESCC development.