胃泌素对胃癌细胞SGC7901 Reg I 基因转录因子的效应.docVIP

  胃泌素对胃癌细胞 SGC7901 Reg I 基因转 录因子的效应# 丁一1，吴艳芳2，张钦宪1** 5 10 15 20 25 30 35 40 45 （1. 郑州大学基础医学院，郑州 450001； 2. 新乡医学院人体解剖学教研室，河南 新乡 453003） 摘要：目的 研究胃泌素对胃癌细胞 SGC7901 Reg I（regenerating gene I，Reg I）基因 转录因子的效应。材料与方法 应用巢式 PCR 技术从胃癌细胞 SGC7901 基因组 DNA 克隆 Reg I 基因启动子 1414bp 片段，序列分析鉴定。应用随机引物法以地高辛分别标记 1414bp 及其 -7 -8 理胃癌细胞 SGC7901 48h，提取核蛋白。应用 Southwestern 印迹技术，分别以地高辛标记 的 1414bp、800bp 和 614bp 片段为探针检测胃泌素对胃癌细胞 SGC7901 Reg I 基因转录因 子的效应。结果 1414bp 探针可检测到 20 条蛋白主带,胃泌素处理后，带 9、12、13、14、 15 和 16 的灰度值明显降低(P＜0.05)。614bp 探针可检测到灰度值变化 6 条主带的带 9、12 和 13，胃泌素处理后，此 3 条主带的灰度值明显降低(P＜0.05)。800bp 探针可检测到灰度 值变化 6 条主带的带 9、12 和 14，胃泌素处理后，仅带 14 的灰度值明显降低(P＜0.05)。 结论 胃癌细胞 SGC7901 Reg I 基因表达由多个转录因子协同调控。降低几个转录因子的结 合活性可能是胃泌素上调胃癌细胞 SGC7901 Reg I 基因表达的途径之一。 关键词：Reg基因；胃泌素；转录因子；胃癌细胞；Southwestern 印迹 中图分类号：R329.1 Effects of gastrin on Reg I gene transcription factors in SGC7901 cell DING Yi1, WU Yanfang2, ZHANG Qinxian1 (1. Histology Embryology, College of Basic Medicine, Zhengzhou University, ZhengZhou 450001; 2. Department of Anatoma, Xinxiang Medical College, HeNan XinXiang 453003) Abstract: IObjective To study the effects of gastrin on Reg I (regenerating gene I） gene transcription factors (TFs) in gastric cancer cell SGC7901. Materials and methods Reg I gene promoter 1414bp fragment was cloned from gastric cancer cell SGC7901 genomic DNA by nest-PCR and identified by sequencing. The 1414bp fragment and its Hind Ⅲ digestion 800bp and 614bp fragments were Digaoxin-labeled by random primer assay and used as probes. Gastric cancer cell SGC7901 was incubated with gastrin-G17 for 48h at final concentrations of 10-7mol/L and 10-8mol/L, and the nuclear proteins were extracted, respectively. Effects of gastrin on Reg I gene TFs were detected by Southwestern blot using Digaoxin-labeled 1414bp, 800bp and 614bp as probes, respectively. Results 20 major bands were detected with 1414bp probe, after gastrin incubation, the densities of band 9, 12, 13, 14, 15 and 16 were decreased significantly (P＜0.05). Of the 6