交通医学

目的：探讨circ-USP14在胃癌中的表达及其对胃癌进展的影响。方法：100例胃癌组织及其邻近的癌旁组织标本，采用PCR方法检测胃癌组织中circ-USP14的表达水平，Kaplan-Meier生存曲线分析circ-USP14高表达组与低表达组患者的生存时间，分析circ-USP14表达与胃癌临床病理特征的关系。培养胃癌细胞株AGS，构建circUSP14过表达质粒，设Vector对照组和circ-USP14过表达组，采用EDU实验检测胃癌细胞增殖活力，Transwell实验检测胃癌细胞迁移能力。从CircInteractome数据库预测可能与circ-USP14结合的miRNA。构建野生型（circ-USP14-wt）与突变型（circ-USP14-mut）的pGL3报告载体，分别与miR-NC和miR-1307共转染至细胞中，采用双荧光素酶检测系统检测circ-USP14与miR-1307的相互作用。Pearson相关性分析胃癌组织中miR-1307与circ-USP14表达的关系。结果：胃癌组织中circ-USP14表达水平较癌旁组织显著下调。Circ-USP14表达水平与肿瘤直径（P=0.003）、TNM分级（P=0.014）、淋巴结转移（P=0.002）密切相关。与Vector组比较，circ-USP14过表达组胃癌细胞增殖活力显著下降，细胞迁移能力明显减弱。双荧光素酶报告实验表明，circ-USP14与miR-1307相互作用，Pearson相关性分析显示胃癌组织中miR-1307与circ-USP14表达呈显著负相关。EDU实验显示转染miR-1307 mimics促进细胞增殖，而circ-USP14过表达抑制miR-1307 mimics的作用。Transwell实验显示miR-1307 mimics促进胃癌细胞迁移，而circUSP14过表达抑制miR-1307 mimics的促进作用。结论：胃癌组织中circ-USP14低表达，与胃癌的恶性生物学行为及患者预后不良有关，circ-USP14通过调控miR-1307表达而抑制胃癌细胞的增殖、迁移能力。

Objective: To explore the expression of circ-USP14 in gastric cancer, and its impact on the development of gastric cancer. Methods: One hundred gastric cancer tissues and their adjacent para-carcinoma tissues were selected.The expression level of circ-USP14 in gastric cancer tissues was assessed by PCR method. Kaplan-Meier survival curve was used to analyze the survival time of patients in the high-expression group and low-expression group of circ-USP14,and the relationship between the expression of circ-USP14 and the clinicopathological characteristics of gastric cancer was analyzed. The gastric cancer cell line AGS was cultivated, and the circ-USP14 overexpression plasmid was constructed.The cells were set as the Vector control group and the circ-USP14 overexpression group. Then, the proliferation ability of gastric cancer cells was detected by the EDU assay, and the migration ability of gastric cancer cells was detected by the Transwell assay. Use the CircInteractome database to predict miRNAs that may interact with circ-USP14. The pGL3 reporter vectors of wild-type(circ-USP14-wt) and mutant(circ-USP14-mut) were constructed and co-transfected with miRNC and miR-1307 into the cells, then the dual luciferase detection system was used to determine the interaction between circ-USP14 and miR-1307. Pearson correlation analysis was used to investigate the relationship between circ-USP14 and miR-1307. Results: The expression level of circ-USP14 in gastric cancer tissues was significantly lower than that in paracarcinoma tissues. The expression level of circ-USP14 was closely related to tumor diameter(P=0.003), TNM grade(P=0.014), and lymph node metastasis(P=0.002). Compared with the Vector group, the proliferation ability of gastric cancer cells in the circ-USP14 overexpression group significantly decreased, and the migration ability was significantly weakened.The dual luciferase reporter assay demonstrated that circ-USP14 interacted with miR-1307. Pearson correlation analysis revealed a significant negative correlation between miR-1307 and circ-USP14 expression in gastric cancer tissues. EDU experiment indicated that transfection of miR-1307 mimics promoted cell proliferation, while overexpression of circ-USP14inhibited the effect of miR-1307 mimics. Transwell assay showed that miR-1307 mimics promoted the migration of gastric cancer cells, but overexpression of circ-USP14 inhibited the promoting effect of miR-1307 mimics. Conclusion: The expression of circ-USP14 in gastric cancer is low, which is associated with the malignant biological behavior of gastric cancer and poor prognosis of patients. Circ-USP14 inhibits the proliferation and migration abilities of gastric cancer cells by regulating miR-1307.