Kruppel样因子4在胃癌细胞中的表达及KLF4基因启动子区的生物信息学分析

摘要

Objective To explore the expression of KLF 4 gene in human gastric cancer cell lines MKN-45 and analyze the gene se-quence of KLF4 promoter region in gastric cancer .Methods Real-time PCR was used to detect KLF4 mRNA expression in MKN-45 and GES-1.A variety of online software were used to analyze the sequences of KLF 4 gene promoter and predict the binding sites of transcriptional factor and relevant transcription factors in gastric cancer .Results Real-time PCR showed that the KLF 4 mRNA level in MKN-45 was signif-icantly lower than that of GES-1(4.24±0.15 vs 6.66±0.23) with significant differences (P<0.01).The promoter region of KLF4 gene was 4 915 bp long,containing 183 transcriptional factors .Among them,17 transcriptional factors were abnormally expressed in gastric cancer ,in-cluding AP-1,AP-2,HIF-1,SP3,etc.Conclusions KLF4 mRNA expression is down-regulated in gastric cancer cell line MKN-45.Bioinfor-matics analysis shows that transcriptional factors AP-1,AP-2,HIF-1 and SP3 may be involved in regulation of KLF4 expression in the devel-opment of gastric cancer ,which provides theoretical basis for further research .%目的：检测Kruppel样因子KLF4基因在人胃癌细胞株MKN-45及人正常胃黏膜上皮细胞株GES-1的表达情况，并应用生物信息学方法分析KLF4基因启动子区序列，预测胃癌发生中其涉及的转录调控因子。方法采用实时荧光定量 PCR 方法检测 MKN-45及 GES-1中 KLF4 mRNA的表达；利用多种在线软件获取并分析人KLF4基因启动子序列，预测其转录因子结合位点和胃癌组织中相关的转录因子。结果实时定量PCR检测结果显示：KLF4 mRNA在人胃癌细胞株MKN-45中的表达低于在人正常胃黏膜细胞株 GES-1中的表达（4.24±0.15 vs 6.66±0.23）（ P＜0.01）；KLF4基因启动子序列全长为4915 bp，共涉及183个转录因子结合位点，在胃癌中异常表达的转录因子有17个，包括AP-1、AP-2、HIF-1、SP3等。结论 KLF4 mRNA在胃癌细胞株MKN-45中表达下调，生物信息学分析提示在胃癌发生中 AP-1、AP-2、HIF-1、SP3转录因子等可能参与调控KLF4的表达。