小细胞肺癌患者与健康人肠道菌群的差异性研究

发布时间：2018-03-26 11:22

  本文选题：小细胞肺癌　切入点：健康人　出处：《大连医科大学》2017年硕士论文

【摘要】：研究背景和目的:小细胞肺癌(Small Cell Lung Cancer,SCLC)约占全部肺癌的14%,它具有高度侵袭性,且生长迅速、倍增时间短、容易出现远处转移、预后差。SCLC确诊时多为广泛期(Extensive disease,ED),其对放化疗均敏感,近期客观缓解率高,但90%以上患者治疗后会在短期内出现复发和转移。人体的肠道内存在着大量的共生微生物,这些共生微生物种类超过1000种[1-2]。许多研究表明,机体肠道的共生微生物能够直接介入宿主的各种代谢进程,发挥了独特且重要的作用,并产生多种益于机体健康的代谢产物[2]。在正常情况时,肠道菌群与机体及外界环境之间保持一种相对平衡状态,来维持机体的健康,若上述平衡状态被打破,就会出现肠道菌群失调,便引起疾病产生[3]。有研究表明肠道菌群失调与多种疾病的发生有关。国内外目前研究多集中在消化道肿瘤与肠道菌群的关系。本研究通过对比初次确诊为SCLC的患者与健康人肠道菌群之间的差异,意从微生态学角度分析SCLC的发病因素,为SCLC的诊断、预防和治疗提供新策略。方法:1、临床粪便标本的收集:筛选大连医科大学附属第二医院肿瘤科14例经病理初次确诊为小细胞肺癌患者,于第一周期化疗前收集新鲜粪便标本一次并标记为1-14号,筛选年龄、性别、吸烟指数、生活地域等因素相匹配的14例健康人,同时收集新鲜粪便标本一次并标记为15-28号,将所收集的标本尽快放置在-80℃冰箱。第一组为1-7号和15-21号标本,第二组为8-14号和22-28号标本。2、肠道细菌DNA的提取:利用粪便DNA提取试剂盒提取小细胞肺癌患者及健康人肠道细菌DNA。3、采用聚合酶链式反应-变性梯度凝胶电泳法(PCR-DGGE)分析肠道菌群的变化:对肠道细菌进行PCR,PCR扩增成功后行DGGE,分析DGGE指纹图谱,切取差异明显条带测序,判断菌属类别。4、结果的分析:运用Quantity One软件及SPSS19.0软件分析所得结果。结果:1、肠道细菌DNA提取及PCR扩增情况:14例初次确诊SCLC患者第一周期化疗前和健康人粪便均提取出肠道细菌DNA,不同粪便标本DNA浓度不同。肠道细菌DNA行PCR扩增后均出现目的条带。2、SCLC组和健康对照组肠道菌群分析:(1)第一组肠道菌群分析:健康对照组的多样性指数及丰富度均高于SCLC组,但是差异不显著(P0.05)。采用非加权法(UPGAMA)进行相似性聚类分析,结果显示,SCLC组和健康对照组未见分成两簇,两组间差异不显著。(2)第二组肠道菌群分析:健康对照组的多样性指数及丰富度均高于SCLC组,但是差异不显著(P0.05)。采用非加权法(UPGAMA)进行相似性聚类分析,结果显示,SCLC组和健康对照组未见分成两簇,两组间差异不显著。(3)SCLC组(1-14号)与健康对照组(15-28号)的肠道菌群分析:健康对照组的多样性指数及丰富度均高于SCLC组,多样性指数有统计学差异(P0.05),丰富度差异不显著(P0.05)。(4)切胶、测序结果分析:切取DGGE图谱上变化明显的条带,1、3、7、8条带在SCLC组中亮度比健康对照组高,2、4、5、6、9条带在健康对照组亮度比SCLC组高。结果显示,条带1、3与嗜木聚糖真杆菌(Eubacteriumxylanophilum)的同源性分别是98%和97%;条带2、6与普氏菌(Prevotella copri)的同源性分别是91%和97%;条带4与布劳特氏菌(Blautia sp.)的同源性是93%;条带5与Dialister succinatiphilus的同源性是92%;条带7与挑剔真杆菌(Eubacterium eligens)的同源性是100%;条带8与梭菌属(Clostridium sp.)的同源性是99%;条带9与瘤胃假丁酸弧菌(Pseudobutyrivibrio ruminis)的同源性是100%。3、SCLC组肠道菌群分析:将14例SCLC患者分别按照分期、年龄、吸烟指数分组并进行统计学处理,结果表明,局限期组的多样性指数及丰富度均低于广泛期组,差异不明显(P0.05);年龄60岁组的多样性指数及丰富度均高于年龄≤60岁组,差异不明显(P0.05);吸烟指数692.857组的多样性指数及丰富度均高于吸烟指数≤692.857组,差异不明显(P0.05)。结论:1.SCLC患者肠道菌群与健康人之间存在差异,多样性差异显著,但丰富度差异不明显。2.嗜木聚糖真杆菌、挑剔真杆菌及梭菌属的增多与SCLC的发生可能存在正相关;普氏菌、瘤胃假丁酸弧菌的减少与SCLC的发生可能存在正相关。3.SCLC患者肠道菌群的结构与其分期、年龄、吸烟指数无显著相关性。
[Abstract]:BACKGROUND & OBJECTIVE : Small Cell Lung Cancer ( NSCLC ) accounts for about 14 % of all lung cancer . It has a high degree of invasion , rapid growth , short doubling time , easy distant metastasis and poor prognosis . There is a large number of commensal microorganisms in the intestinal tract of human body . There are a lot of symbiotic microorganisms in the intestinal tract of human body . Many studies show that the symbiotic microorganisms in the body of the body can directly intervene in various metabolic processes of the host , play a unique and important role , and produce a variety of metabolic products beneficial to the health of the organism . in that normal condition , the intestinal flora and the organism and the external environment maintain a relatively balanced state to maintain the health of the organism . The results were as follows : 1 . The results were analyzed by PCR - DGGE . The results were as follows : 1 . The results were as follows : 1 - 7 and 15 - 21 . The results showed that 1 - 7 and 15 - 21 samples were used for the diagnosis , diagnosis , prevention and treatment . The results showed that the homology of bands 1 , 3 and Prevotella copri was 91 % and 97 % , and bands 2 , 6 and Prevotella copri were 91 % and 97 % , respectively . The homology was 93 % ; the homologies of bands 5 and dial were 92 % ; the homology between bands 7 and Eubacterium eligens was 100 % ; bands 8 and Clostridium sp . The results showed that the diversity index and the richness of the group were significantly higher than those in the age 鈮，

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