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Objective: To investigate the role of human umbilical cord mesenchymal stem cell (hucMSC)-derived exosomes in the Wnt signaling pathway and their effects on fracture healing in rats., Materials and Methods: A total of 24 healthy male Sprague-Dawley (SD) rats were randomly divided into 3 groups, of which the experimental groups were injected with Phosphate-Buffered Saline (PBS) and hUCMSC-derived exosomes, respectively, at the fracture site, and a blank control group was set. At 2 and 3 w after treatment, respectively, the healing condition at the fracture site in the rats was detected by micro-computed tomography (CT). The protein expressions of β-catenin and Wnt3a of the Wnt signaling pathway in the bone tissue were measured via Western blotting (WB) assay. Quantitative Real Time-fluorescence Polymerase Chain Reaction (qRT-PCR) was performed to determine the expressions of osteogenic marker genes [collagen type I (COL-1), osteopontin (OPN) and runt-related transcription factor 2 (RUNX2)]., Results: The results of the micro-CT scan showed that the rats treated with exosomes had better apposition of the fracture site, and the appearance of cortical bone was continuous. The fracture sites in the blank control group and PBS injection group were not healed, and the appearance of cortical bone was discontinuous, with significant fracture lines. According to the WB results, the protein expression levels of β-catenin and Wnt3a in exosome treatment group were significantly higher than those in the blank control group and PBS injection group (p<0.01). The qRT-PCR results indicated that the expression levels of COL-1, OPN and RUNX2 in exosome treatment group were increased evidently compared with those in the other two groups (p<0.01)., Conclusions: HucMSC-derived exosomes are probably involved in the repair of fracture in rats through the Wnt signaling pathway.

Objective: To explore the expression of extracellular vesicle-derived lncZEB1-AS1 in esophageal cancer and its role in esophageal cancer progression., Patients and Methods: The extracellular vesicles (EVs) from esophageal cancer patients (n = 26) and normal subjects (n = 26) were isolated by differential centrifugation. The expression of lncZEB1-AS1 in EVs was detected by Real-time PCR (polymerase chain reaction). The clinical data of normal subjects and patients were analyzed. In addition, the concentration of EVs and lncZEB1-AS1 in blood samples from normal subjects and esophageal cancer patients were assessed. After co-culture of esophageal cancer cell line EC109 and EVs with or without lncZEB1-AS1 knockdown, cell proliferation was detected by CCK-8 assay. The possible target microRNAs of lncZEB1-AS1 in cytoplasm were predicted with miRcode, followed by correlation analysis of lncZEB1-AS1 and miR-214. Through literature review, lncZEB1-AS1 positively regulates ZEB1 expression, which was consistent with our result., Results: Quantitative Real-time PCR showed that the serum levels of EVs and the content of lncZEB1-AS1 in EVs from esophageal cancer patients were significantly higher than those in normal controls. LncZEB1-AS1 was overexpressed in esophageal cancer cells co-cultured with EVs of esophageal cancer patients. CCK-8 results indicated that EC109 cells co-cultured with EVs of esophageal cancer patients had stronger proliferative capacity. miRcode showed that miR-214 ranked the first of microRNAs that lncZEB1-AS1 might target, and miR-214 expression was significantly increased after lncZEB1-AS1 knockdown in EC109. After overexpressing lncZEB1-AS1 in EC109 or co-culturing EVs of esophageal cancer patients with EC109 cells, we found that lncZEB1-AS1 positively regulates ZEB1. In contrast, interfering with the expression of lncZEB1-AS1 in esophageal cancer cell lines can effectively reduce the expression of ZEB1., Conclusions: EVs in the peripheral blood from esophageal cancer patients promote esophageal cancer progression by delivering lncZEB1-AS1 to esophageal cancer cells and targeting miR-214.