發(fā)布時(shí)間：2018-04-24 14:00

  本文選題：惡性淋巴瘤 + T細(xì)胞淋巴瘤��； 參考：《福建醫(yī)科大學(xué)》2008年碩士論文

【摘要】： 【目的】 通過腹腔注射DNA烷化劑N-甲基-N-亞硝基脲(MNU)誘導(dǎo)小鼠胸腺淋巴瘤,建立相應(yīng)動(dòng)物模型;應(yīng)用組織形態(tài)學(xué)、免疫表型、超微結(jié)構(gòu)觀察及巢式聚合酶鏈反應(yīng)(PCR)方法鑒定腫瘤細(xì)胞來(lái)源并分型。 【方法】 1.第一部分,選用6～8周齡C57BL/6小鼠52只,隨機(jī)分實(shí)驗(yàn)組40只和對(duì)照組12只。實(shí)驗(yàn)組小鼠腹膜腔內(nèi)注射新鮮配制MNU誘導(dǎo)液(50mg/kg體重);對(duì)照組腹膜腔內(nèi)注射等量生理鹽水,共注射2次(分別于第0、8周)。注射后觀察動(dòng)物一般情況,對(duì)瀕死及死亡小鼠進(jìn)行解剖觀察。第22周采用頸椎脫位法處死全部小鼠,解剖檢查胸腺淋巴瘤的發(fā)生及其他臟器情況。 2.第二部分,應(yīng)用光鏡和透射電鏡,對(duì)誘發(fā)的胸腺淋巴瘤及其侵襲臟器進(jìn)行研究,分析其組織學(xué)及超微結(jié)構(gòu)特點(diǎn)。通過免疫組織化學(xué)方法(所選抗體有CD3、CD20、TdT及PCNA)鑒定腫瘤細(xì)胞來(lái)源、分型。 3.第三部分,采集8例實(shí)驗(yàn)組胸腺淋巴瘤組織及4例對(duì)照組胸腺組織,迅速放入液氮罐內(nèi),后置入-70℃冰箱凍存。提取基因組DNA,采用巢式PCR方法,對(duì)采集樣本進(jìn)行T細(xì)胞受體β鏈(TCRβ)和γ鏈(TCRγ)克隆性基因重排分析,并對(duì)TCRγ基因重排的PCR產(chǎn)物直接測(cè)序。 【結(jié)果】 1.實(shí)驗(yàn)后期,實(shí)驗(yàn)組小鼠體重明顯下降。第22周,67.5%(27?40)實(shí)驗(yàn)組小鼠產(chǎn)生胸腺淋巴瘤。不同性別對(duì)成瘤率影響無(wú)明顯差異。77.8%胸腺淋巴瘤合并其他臟器侵犯。 2. 27例胸腺淋巴瘤光鏡下主要表現(xiàn)為:胸腺結(jié)構(gòu)破壞,代之以彌漫分布的中等大小淋巴樣細(xì)胞,細(xì)胞形態(tài)比較一致,胞質(zhì)稀少,核圓形、卵圓形或扭曲核,染色質(zhì)細(xì)膩,核仁顯著,核分裂象多見,可見“星空”現(xiàn)象。免疫組化顯示瘤細(xì)胞表達(dá)CD3和TdT。超微結(jié)構(gòu)觀察,瘤細(xì)胞平均直徑7.02±1.12μm,表面無(wú)突起,核形相對(duì)規(guī)則或伴有扭曲核,核仁靠近核膜,核漿比例高,胞質(zhì)內(nèi)細(xì)胞器極少,凋亡細(xì)胞易見。 3. 8例胸腺淋巴瘤檢測(cè)TCRβ和TCRγ均呈克隆性基因重排,陽(yáng)性率100%;4例對(duì)照組胸腺組織均未檢測(cè)到TCRβ或TCRγ克隆性基因重排。DNA序列測(cè)定證實(shí)TCRγ基因PCR擴(kuò)增產(chǎn)物為基因重排產(chǎn)物。 【結(jié)論】 1.腹腔分次注射MNU可以誘導(dǎo)小鼠產(chǎn)生胸腺淋巴瘤,其生物學(xué)行為與人類腫瘤相似。 2.結(jié)合光鏡、免疫組化和電鏡檢查結(jié)果,證實(shí)所有腫瘤為胸腺T淋巴細(xì)胞起源的前驅(qū)淋巴母細(xì)胞淋巴瘤。 3.巢式PCR TCRβ和TCRγ的基因重排檢測(cè)及DNA序列分析證實(shí),MNU誘導(dǎo)的小鼠胸腺淋巴瘤實(shí)際上是T淋巴細(xì)胞的單克隆性增生,是來(lái)源于T細(xì)胞的腫瘤。 總之,MNU誘導(dǎo)的小鼠胸腺淋巴瘤在生物學(xué)行為、形態(tài)學(xué)、免疫表型和遺傳學(xué)方面均與人類相應(yīng)腫瘤極其相似,可作為人類這一腫瘤實(shí)體的理想動(dòng)物模型。
[Abstract]:[purpose] Mouse thymic lymphoma was induced by intraperitoneal injection of DNA alkylator N-methyl-N-nitroso (MNUN), and the corresponding animal model was established by using histomorphology and immunophenotype. Ultrastructural observation and nested polymerase chain reaction (PCR) were used to identify the origin and type of tumor cells. [methods] 1. In the first part, 52 C57BL/6 mice aged 6 and 8 weeks were randomly divided into experimental group (n = 40) and control group (n = 12). The mice in the experimental group were injected with 50 mg / kg body weight of fresh MNU inducer in peritoneal cavity and the control group were injected with the same amount of normal saline twice (at the 0th week of 8 weeks respectively). The animals were observed after injection, and the near-death and dead mice were dissected and observed. All the mice were killed by cervical dislocation at the 22nd week. The occurrence of thymic lymphoma and other organs were dissected. 2. In the second part, the histological and ultrastructural characteristics of thymic lymphoma and its invasive organs were studied with light microscope and transmission electron microscope. The origin and typing of tumor cells were identified by immunohistochemical method (CD3, CD20, TdT and PCNA). 3. In the third part, 8 cases of thymic lymphoma in experimental group and 4 cases of thymic tissue in control group were collected and put into liquid nitrogen tank quickly, then frozen in -70 鈩，

本文編號(hào)：1796936