本文選題：小細(xì)胞肺癌　切入點(diǎn)：健康人　出處：《大連醫(yī)科大學(xué)》2017年碩士論文

【摘要】：研究背景和目的:小細(xì)胞肺癌(Small Cell Lung Cancer,SCLC)約占全部肺癌的14%,它具有高度侵襲性,且生長(zhǎng)迅速、倍增時(shí)間短、容易出現(xiàn)遠(yuǎn)處轉(zhuǎn)移、預(yù)后差。SCLC確診時(shí)多為廣泛期(Extensive disease,ED),其對(duì)放化療均敏感,近期客觀緩解率高,但90%以上患者治療后會(huì)在短期內(nèi)出現(xiàn)復(fù)發(fā)和轉(zhuǎn)移。人體的腸道內(nèi)存在著大量的共生微生物,這些共生微生物種類超過(guò)1000種[1-2]。許多研究表明,機(jī)體腸道的共生微生物能夠直接介入宿主的各種代謝進(jìn)程,發(fā)揮了獨(dú)特且重要的作用,并產(chǎn)生多種益于機(jī)體健康的代謝產(chǎn)物[2]。在正常情況時(shí),腸道菌群與機(jī)體及外界環(huán)境之間保持一種相對(duì)平衡狀態(tài),來(lái)維持機(jī)體的健康,若上述平衡狀態(tài)被打破,就會(huì)出現(xiàn)腸道菌群失調(diào),便引起疾病產(chǎn)生[3]。有研究表明腸道菌群失調(diào)與多種疾病的發(fā)生有關(guān)。國(guó)內(nèi)外目前研究多集中在消化道腫瘤與腸道菌群的關(guān)系。本研究通過(guò)對(duì)比初次確診為SCLC的患者與健康人腸道菌群之間的差異,意從微生態(tài)學(xué)角度分析SCLC的發(fā)病因素,為SCLC的診斷、預(yù)防和治療提供新策略。方法:1、臨床糞便標(biāo)本的收集:篩選大連醫(yī)科大學(xué)附屬第二醫(yī)院腫瘤科14例經(jīng)病理初次確診為小細(xì)胞肺癌患者,于第一周期化療前收集新鮮糞便標(biāo)本一次并標(biāo)記為1-14號(hào),篩選年齡、性別、吸煙指數(shù)、生活地域等因素相匹配的14例健康人,同時(shí)收集新鮮糞便標(biāo)本一次并標(biāo)記為15-28號(hào),將所收集的標(biāo)本盡快放置在-80℃冰箱。第一組為1-7號(hào)和15-21號(hào)標(biāo)本,第二組為8-14號(hào)和22-28號(hào)標(biāo)本。2、腸道細(xì)菌DNA的提取:利用糞便DNA提取試劑盒提取小細(xì)胞肺癌患者及健康人腸道細(xì)菌DNA。3、采用聚合酶鏈?zhǔn)椒磻?yīng)-變性梯度凝膠電泳法(PCR-DGGE)分析腸道菌群的變化:對(duì)腸道細(xì)菌進(jìn)行PCR,PCR擴(kuò)增成功后行DGGE,分析DGGE指紋圖譜,切取差異明顯條帶測(cè)序,判斷菌屬類別。4、結(jié)果的分析:運(yùn)用Quantity One軟件及SPSS19.0軟件分析所得結(jié)果。結(jié)果:1、腸道細(xì)菌DNA提取及PCR擴(kuò)增情況:14例初次確診SCLC患者第一周期化療前和健康人糞便均提取出腸道細(xì)菌DNA,不同糞便標(biāo)本DNA濃度不同。腸道細(xì)菌DNA行PCR擴(kuò)增后均出現(xiàn)目的條帶。2、SCLC組和健康對(duì)照組腸道菌群分析:(1)第一組腸道菌群分析:健康對(duì)照組的多樣性指數(shù)及豐富度均高于SCLC組,但是差異不顯著(P0.05)。采用非加權(quán)法(UPGAMA)進(jìn)行相似性聚類分析,結(jié)果顯示,SCLC組和健康對(duì)照組未見(jiàn)分成兩簇,兩組間差異不顯著。(2)第二組腸道菌群分析:健康對(duì)照組的多樣性指數(shù)及豐富度均高于SCLC組,但是差異不顯著(P0.05)。采用非加權(quán)法(UPGAMA)進(jìn)行相似性聚類分析,結(jié)果顯示,SCLC組和健康對(duì)照組未見(jiàn)分成兩簇,兩組間差異不顯著。(3)SCLC組(1-14號(hào))與健康對(duì)照組(15-28號(hào))的腸道菌群分析:健康對(duì)照組的多樣性指數(shù)及豐富度均高于SCLC組,多樣性指數(shù)有統(tǒng)計(jì)學(xué)差異(P0.05),豐富度差異不顯著(P0.05)。(4)切膠、測(cè)序結(jié)果分析:切取DGGE圖譜上變化明顯的條帶,1、3、7、8條帶在SCLC組中亮度比健康對(duì)照組高,2、4、5、6、9條帶在健康對(duì)照組亮度比SCLC組高。結(jié)果顯示,條帶1、3與嗜木聚糖真桿菌(Eubacteriumxylanophilum)的同源性分別是98%和97%;條帶2、6與普氏菌(Prevotella copri)的同源性分別是91%和97%;條帶4與布勞特氏菌(Blautia sp.)的同源性是93%;條帶5與Dialister succinatiphilus的同源性是92%;條帶7與挑剔真桿菌(Eubacterium eligens)的同源性是100%;條帶8與梭菌屬(Clostridium sp.)的同源性是99%;條帶9與瘤胃假丁酸弧菌(Pseudobutyrivibrio ruminis)的同源性是100%。3、SCLC組腸道菌群分析:將14例SCLC患者分別按照分期、年齡、吸煙指數(shù)分組并進(jìn)行統(tǒng)計(jì)學(xué)處理,結(jié)果表明,局限期組的多樣性指數(shù)及豐富度均低于廣泛期組,差異不明顯(P0.05);年齡60歲組的多樣性指數(shù)及豐富度均高于年齡≤60歲組,差異不明顯(P0.05);吸煙指數(shù)692.857組的多樣性指數(shù)及豐富度均高于吸煙指數(shù)≤692.857組,差異不明顯(P0.05)。結(jié)論:1.SCLC患者腸道菌群與健康人之間存在差異,多樣性差異顯著,但豐富度差異不明顯。2.嗜木聚糖真桿菌、挑剔真桿菌及梭菌屬的增多與SCLC的發(fā)生可能存在正相關(guān);普氏菌、瘤胃假丁酸弧菌的減少與SCLC的發(fā)生可能存在正相關(guān)。3.SCLC患者腸道菌群的結(jié)構(gòu)與其分期、年齡、吸煙指數(shù)無(wú)顯著相關(guān)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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