囊型包虫病患者包囊钙化相关基因研究  

Study of the genes correlated with cyst calcification in patients with cystic echinococcosis

作　　者：尹媛婕 张强[2] 杨银学[3] 杨书鲲[4] 王海丰 施锦绣 王志敏 杨延辉[1] 林源[1] 李昭宇[3] 杨玉荣[1] 

机构地区：[1]宁夏医科大学基础医学院,银川750004 [2]山东菏泽市立医院胃肠外科 [3]宁夏医科大学总医院普外科 [4]宁夏医科大学第二附属医院影像科 [5]上海南方科技有限公司 [6]上海人类基因组研究中心遗传部

出　　处：《中华预防医学杂志》2016年第5期434-438,共5页Chinese Journal of Preventive Medicine

基　　金：国家自然科学基金（30960339、81460311）

摘　　要：目的探寻与囊型包虫病患者包囊钙化相关的基因，从分子生物学水平提供判断患者预后情况的依据。方法于2013年6月至2014年12月，收集前往宁夏3家医院就诊的32例囊型包虫病患者（10例进行mRNA基因芯片分析，22例进行Real—timePCR分析）和11例肝囊腺瘤患者的肝组织。采用基因表达差异显著性分析方法对10例囊型包虫病患者（钙化组与非钙化组各5例）的基因表达差异进行分析和筛选，用倍数变化衡量同一基因在两组之间表达量的变化情况。采用Real-timePCR在22例囊型包虫病患者（钙化组与非钙化组各11例）之间进行验证，以GAPDH作为内参基因，并用11例肝囊腺瘤患者进行标化，计算钙化组和非钙化组之间半乳糖凝集素4（LGALS4）和酸性神经酰胺酶（ASAHl）的相对表达量，并采用r检验比较两组的差异。结果经基因表达谱芯片筛选出5个表达有显著性差异的基因，且都在钙化组中表达下调，其中LGALS4和ASAHl下调程度较明显，倍数变化分别为0．0088和0．0203。Real．timePCR验证结果显示，LGALS4在钙化组的相对表达量为0．49±0．27，非钙化组为2．70±2．61，差异有统计学意义（t=-2．59，P=0．026）；ASAHl在钙化组的相对表达量为1．36±0．33，非钙化组为1．68±0．67，差异无统计学意义（t=-1．44，P=0．167）。在非钙化组中，LGALS4和ASAHl的表达量波动范围较大，且两者表达量变化的趋势呈正相关（r=O．91，P=0．001），即LGALS4表达量较低的患者其ASAHl表达量也相对较低。结论LGALS4和ASAHl在发生钙化的囊型包虫病患者中低表达，可能成为判断囊型包虫病自愈倾向的标志基因。Objective To explore the genes （biomarkers） correlated with cyst calcification in patients with cystic eehinoeoeeosis （CE）, and to provide the evidence for the judgment on the patients＇ prognosis at molecular level. Methods The liver tissues from 32 patients with liver CE （10 cases for mRNA microarray and 22 cases for real-time PCR analysis） and 11 patients with hepatic cystadenoma were collected from three hospitals in Ningxia from June, 2013 to December, 2014. A comparison of the different gene-expressions between five patients with calcified lesions and five cases with no calcification was carried out using Significant Analysis of Microarrays （SAM） to select a subset of differentially expressed genes （DEGs） . Fold-change analysis was used to assess the changes of the expression quantity in the same genes between two groups. The verification was conducted among the liver tissues from 22 patients with liver CE （11 in the group of calcified or 11 in that of non-calcified） by real-time quantitative PCR （RT-qPCR）. With GAPDH as a reference-gene and the liver tissues from 11 cases with hepatic cystadenoma as standardized control groups, the relative expressions of galecitin-4 （LGALS4） and acid eeramidase （ASAH1） in patients with calcified and non-calcified were calculated, respectively. The differences between two groups were compared using t＇-test. Results Five screened genes presented siginifieantly different expressions all had showed the low-regulated expressions in the calcified group, with the most distinct low-regulation of LGALS4 and ASAH1 whose fold changes were 0.008 8, and 0.020 3, respectively. The verification by RT-qPCR illustrated that the relative expression of LGALS4 was showed at level of 0.49 ＋ 0.27 amongst patients with calcified, and at level of 2.70 - 2.61 amongst non-calcified individuals,indicating significant differences between two groups （t=-2.59, P=0.026）; while the ASAH1 was relatively expressed at levels of 1.36 ＋ 0.33 and of 1.68 ± 0.67

关 键 词：包虫病 基因表达谱 半乳糖凝集素 酸性神经酰胺酶 

分 类 号：R532.32[医药卫生—内科学]