【摘要】 目的研究S1P5对食管鳞癌Eca109细胞黏附能力的影响。方法从人正常食管黏膜上皮克隆S1P5基因,构建表达载体S1P5-EGFP,转染Eca109细胞,经G418筛选,获得S1P5-EGFP/Eca109稳定细胞株。以对照载体Control-EGFP转染的Eca109细胞作为对照,荧光显微镜下观察细胞的形态,用黏附试验评价其黏附能力。结果成功构建S1P5-EGFP和Control-EGFP载体并转染Eca109细胞获得稳定细胞株。S1P5表达于细胞膜并使细胞略呈梭形改变。S1P5过表达致Eca109细胞黏附能力明显低于对照(P<0.05),并且不受其配体1-磷酸鞘氨醇(S1P)的影响。结论 S1P5组成性抑制Eca109细胞的黏附能力,食管癌细胞可能通过下调S1P5的表达增强其黏附。更多还原

【Abstract】 Objective To investigate the effect of S1P5 on adhesion of human esophageal squamous cell carcinoma(ESCC) cell line Eca109.Methods Human S1P5 gene was cloned from normal esophageal mucosal epithelium.S1P5-EGFP vector was constructed and transfected into Eca109 cells.The S1P5 expressing Eca109 cell was designated S1P5-EGFP/Eca109.With Control-EGFP vector transfecting Eca109 cell as control,cell morphology and adhesion were analyzed by fluorescence microscopy and cell adhesion assay,respectively.Results S1P5-EGFP and Control-EGFP vectors,and their corresponding stably-transfected Eca109 cells were successfully constructed.S1P5 overexpressing Eca109 cells displayed spindle cell morphology and S1P5 was expressed on the plasma membrane.The cell adhesion of S1P5-transfected Eca109 cells was lower than control vector-transfected cells with or without sphingosine 1-phosphate(P<0.05).Conclusion The data demonstrate that S1P5 constitutively inhibits Eca109 cell adhesion.Esophageal cancer cells may down-regulate the expression of S1P5 to promote adhesion.更多还原