摘要
目的检测Hedgehog蛋白及基因甲基化水平在正常食管黏膜（N）、食管不典型增生（HGD）及食管癌（EC）中的表达,探讨Hedgehog通路在食管癌发病机制中的作用.方法收集手术及内镜下切除的N、食管轻度不典型增生（LGD）、HGD及EC组织标本,应用免疫组织化学染色、蛋白印迹法（Western-blot）及实时定量PCR（RT-PCR）检测Hedgehog蛋白及基因的表达,并用甲基化特异性PCR检测Hedgehog基因的甲基化水平.结果Sonic Hedgehog（SHh）蛋白在N及LGD、HGD及EC的表达差异均无统计学差异（LGD比N：t=1.96,P=0.67;HGD比EC：t=1.59,P=0.53）;在HGD及EC中的表达均高于N及LGD（HGD比N：t=0.593,P=0.004;HGD比LGD：t=1.308,P=0.325,EC比N：t=0.292,P=0.000;EC比LGD：t=0.734,P=0.004）;Hedgehog基因在EC中的表达高于N、LGD及HGD（EC比N：t=0.909,P=0.019;EC比LGD：t=0.398,P=0.007;EC比HGD：t=0.843,P=0.012）.Hedgehog基因甲基化水平在EC中明显低于N、LGD及HGD（EC比N：t=0.034,P=0.000;EC比LGD：t=0.102,P=0.000;EC比HGD：t=0.367,P=0.018）.结论Hedgehog可能作为癌基因参与了EC的发生发展,而Hedgehog基因去基化可能是其EC中活化的机制之一,有必要进一步研究探讨其作为EC潜在的基因治疗靶点.
Objective To explore the role of the Hedgehog singaling in the pathogenesis of esophageal cancer.Methods Tissue samples of normal esophageal mucosa,low-grade dysplasia,high-grade dysplasia and esophageal cancer were collected.Immunohistochemical stain,Western-blot and RT-PCR were employed to detect theexpression of Hedgehog protein and gene,methylation special PCR was used to detect the methylation status of Hedgehog gene.Results There was no difference in the expression of SHh protein among nornaml esophageal and low-grade dysplasia,high-grade dysplasia and esophageal cancer（LGD vs.N：t=1.96,P=0.67;HGD vs.EC：t=1.59,P=0.53）.However,the expression of SHh protein in high-grade dysplasia and esophageal cancer was higher than that in normal esophageal mucosa and low-grade dysplasia（HGD vs.N：t=0.593,P=0.004;HGD vs.LGD：t=1.308,P=0.325;EC vs.N：t=0.292,P=0.000;EC vs.LGD：t=0.734,P=0.004）.The expression of Hedgehog gene in esophageal cancer was higher than that in normal esophageal mucosa,low-grade dysplasia and high-grade dysplasia（EC vs.N：t=0.909,P=0.019,EC vs.LGD：t=0.398,P=0.007;EC vs.HGD：t=0.843,P=0.012）.The gene methylation in esophageal cancer was lower than that in normal esophageal mucosa,low-grade dysplasia and high-grade dysplasia（EC vs.N：t=0.0340,P=0.000;EC vs.LGD：t=0.102,P=0.000;EC vs.HGD：t=0.367,P=0.018）.Conclusion Hedgehog signaling may play a role in the pathogenesis and development of esophageal cancer,however,demethylation of Hedgehog gene may be one of the mechanism that cause the activity of oncogene in esophageal cancer.
出处
《中国基层医药》
CAS
2017年第16期2506-2509,J0007,共5页
Chinese Journal of Primary Medicine and Pharmacy
