血红素加氧酶1基因启动子多态性与食管鳞状细胞癌的关系  

The risk of esophageal squamous cell carcinoma associated with microsatellite polymorphism in promoter of heme oxygenase-1

作　　者：胡炳川 张健[1] 王佶[1] 朱成楚[1] 陈保富[1] 孔敏[1] 张波[1] 朱敏[2] 

机构地区：[1]温州医科大学附属浙江省台州医院心胸外科, 浙江省临海市317000 [2]温州医科大学附属浙江省台州医院公共实验平台, 浙江省临海市317000

出　　处：《中华外科杂志》2015年第3期206-210,共5页Chinese Journal of Surgery

基　　金：浙江省重大科技专项资助项目（2011C13039-2）

摘　　要：目的 分析人血红素加氧酶1（HO-1）基因启动子区（GT）n微卫星多态性与汉族人群患食管鳞状细胞癌的风险及相关临床病理学特征的关联性,探讨（GT）n多态性与食管癌的关系.方法 选取2011年1月至2013年12月年龄为40～ 79岁的126例食管鳞状细胞癌患者为病例组,其中男性83例,女性43例,平均年龄（6l＆#177;8）岁;选择同期健康体检人群134名为对照组.对两组的血液标本应用PCR技术扩增（GT）n特定片段,并进行基因测序,根据（GT）n的重复次数把其分为S（n＜25）、M（25≤n＜30）、L（n≥30）3类等位基因,统计其频率分布情况,x2检验评价多态性与食管鳞状细胞癌及临床病理特征的关系.构建相应的启动子荧光素酶报告质粒,转染食管鳞状细胞癌细胞株Ecal09,检测相对荧光素酶活性比值,方差分析评价不同长度的（GT）n其启动子活性情况.结果 病例组L型等位基因频率（25.8％比14.9％,x2=9.520,P=0.002）、L型等位基因携带率高于对照组（41.3％比27.6％,x2=5.381,P=0.020）.L型等位基因携带患者较非L型等位基因携带患者有较高的淋巴结转移率（63.5％比41.8％,x2=5.685,P=0.017）和低分化鳞状细胞癌细胞检出率（53.8％比28.4％,x2=8.335,P=0.004）.H202处理组中,转染携带（GT）16重组报告质粒的Ecal09细胞株相对荧光素酶活性较双蒸水处理组增加（F=23.615,P=0.008）;在双蒸水处理组中,转染携带（GT）16重组报告质粒的细胞相对荧光素酶活性较（GT）26和（GT）36增加（F =41.376,P=0.003：F=50.761,P=0.002）.结论 携带HO-1基因启动子长（GT）n重复序列（n≥30）可能增加食管鳞状细胞癌的易感性并加大淋巴结转移的风险.Objective To investigate the relationship between （GT） n polymorphism and esophageal cancer by analyzing the connection between microsatellite polymorphisms in the promoter of heme oxygenase1 and the clinicopathological characteristics of esophageal squamous celt carcinoma （ESCC） in Han chinese population.Methods The （GT） n repeats in HO-1 gene in 83 male and 43 female hospital-based patients with ESCC （aged between 40 and 79 years with a mean of （61 ＆#177; 8） years） and 134 healthy control individuals were obtained by DNA sequencing.Polymorphisms of the （GT） n repeats were generally grouped into three classes based on allele frequencies：class S alleles （〈 25 repeats）,class M alleles （25 to 29 repeats）,and class L alleles （≥ 30 repeats）.The correlation between susceptibility and clinicopathological characteristics of ESCC were analyzed by x2 test.For in vitro experiments,the transient-transfection assay was performed to explore the correlation between different lengths of （GT） n repeats and promoter activity by assessing the promoter activities of HO-1 gene in cultured Ecal09 cells treated with H2O2 by analysis of cariance.Results Higher frequencies of L-allele （25.8% vs.14.9%,x2 =9.520,P =0.002）,L-allele carrier （41.3% vs.27.6%,x2 =5.381,P =0.020） were found in patients with ESCC.Furthermore,the lymphatic metastasis rate （63.5% vs.41.8%,x2 =5.685,P =0.017） and the detection rate of poorly differentiated ESCC cell （53.8% vs.28.4%,x2 =8.335,P =0.004） was significantly higher in L-allele carriers compared to non-L-allele carriers.In transfection experiments,promoter activities of 5＇-flanking regions of the HO-1 gene in Eca109 cells transfected with the recombinant gene carrying （GT） 16 repeat after treatment with H2 O2 increased （F =23.615,P =0.008）.In H2 O treated control group,compared to （GT） 26 ant （GT）36,the basal promoter activities of HO-l gene carrying （GT） 16 repeat increased （F =41.376,P =0.003 ; F =50.761,P =0.002）.

关 键 词：食管肿瘤 血红素加氧酶-1 多态性 限制性片段长度 疾病遗传易感性 

分 类 号：R735.1[医药卫生—肿瘤]