非小细胞肺癌Hedgehog信号通路关键调节因子HHIP基因甲基化状态检测的研究  

Detection of HHIP gene methylation state of Hedgehog signal pathway in non-small cell lung cancer

作　　者：许靖[1] 张舒[1] 肖中[1] 王海涛[1] 蔡峰[1] Xu Jing;Zhang Shu;Xiao Zhong;Wang Haitao;Cai Feng(Department of Respiratory Medicine, the Seventh People′s Hospital affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200137, China)

机构地区：[1]上海中医药大学附属第七人民医院呼吸内科,200137

出　　处：《国际呼吸杂志》2019年第3期167-171,共5页International Journal of Respiration

基　　金：上海市浦东新区科技发展基金项目(PKJ2015-Y12).

摘　　要：目的采用甲基化基因检测技术对肺癌组织HHIP基因启动子区CpG岛甲基化状态进行分析。方法采用甲基化特异性聚合酶链式反应和亚硫酸盐测序聚合酶链式反应技术,对在上海中医药大学附属第七人民医院接受手术治疗的非小细胞肺癌患者的肿瘤组织及距离肿瘤5cm的癌旁组织进行HHIP启动子区甲基化分析,并使用实时荧光定量聚合酶链式反应检测HHIP基因及Hedgehog信号通路PTCH、GLI1、SHH基因的mRNA表达。结果肿瘤组织HHIP基因启动子区CpG位点部分甲基化,甲基化程度为(3.36±2.35)%,而癌旁组织HHIP基因启动子区CpG位点少数甲基化,甲基化程度为(0.85±0.27)%,差异有统计学意义(t=-3.349,P=0.008)。肿瘤组织的HHIPmRNA相对表达量明显低于癌旁组织(Z=-4.038,P<0.001),其下游PTCH、GLI1、SHH基因的mRNA相对表达量明显高于癌旁组织(Z值均为-4.038,P值均<0.001)。结论在非小细胞肺癌中,抑癌基因HHIP启动子呈甲基化状态并出现表达沉默或下调,可作为肺癌的早期预测指标。Objective To detect the methylation state of CpG island in HHIP gene promoter region of lung cancer tissue by methylation detection technique. Methods By methylation-specific polymerase chain reaction and sulfite sequencing polymerase chain reaction techniques, HHIP methylation status was analyzed in tumor and adjacent tissues of non-small cell lung cancer patients treated by surgery in the Seventh People′s Hospital affiliated to Shanghai University of Traditional Chinese Medicine.The mRNA expressions of HHIP gene and Hedgehog signal pathway PTCH gene, GLI1 gene and SHH gene were detected by real-time fluorescence quantitative polymerase chain reaction. Results The methylation of HHIP gene promoter region CpG loci in the tumor tissues was (3.36±2.35)%, while in the adjacent tissues was (0.85±0.27)% (t=-3.349, P=0.008). The expression of HHIP gene in the tumor tissues was significantly lower than that in the adjacent tissues (Z=-4.038, P<0.001). The relative mRNA expressions of PTCH, GLI1 and SHH genes in tumor tissues were significantly higher than those in adjacent tissues (all Z=-4.038, all P<0.001). Conclusions In non-small cell lung cancer, the promoter of HHIP is methylated and expression is silenced or down-regulated, which can be used as an early predictor of lung cancer.

关 键 词：癌 非小细胞肺 甲基化 HEDGEHOG通路 HHIP基因 

分 类 号：R734.2[医药卫生—肿瘤]