【摘要】：目的通過建立小鼠炎癥性腸病乳腺癌肺轉(zhuǎn)移模型,觀察炎癥性腸病是否可加速乳腺癌的肺部轉(zhuǎn)移過程,探索其可能機(jī)制,以期為臨床上防治乳腺癌合并炎癥性腸病患者發(fā)生腫瘤遠(yuǎn)處轉(zhuǎn)移提供新的干預(yù)策略。方法1依據(jù)實(shí)驗(yàn)室前期實(shí)驗(yàn)動(dòng)物造模方法,以Balb/c雌鼠為實(shí)驗(yàn)對(duì)象,建立2.5%葡聚糖硫酸鈉(dextran sulfate sodium,DSS)誘導(dǎo)的炎癥性腸病(inflammatory bowel disease,IBD)模型,并在此模型基礎(chǔ)上通過尾靜脈接種小鼠乳腺癌細(xì)胞4T1 100ul(1×106/ml)建立IBD乳腺癌肺轉(zhuǎn)移動(dòng)物模型,動(dòng)物實(shí)驗(yàn)結(jié)果重復(fù)2次,每次實(shí)驗(yàn)動(dòng)物中每組動(dòng)物至少10只(每組分別選取5只動(dòng)物進(jìn)行統(tǒng)計(jì)學(xué)分析)。2通過結(jié)腸組織病理切片評(píng)價(jià)IBD乳腺癌肺轉(zhuǎn)移模型中的結(jié)腸炎癥情況,通過肺組織病理切片觀察IBD乳腺癌肺轉(zhuǎn)移模型中的肺轉(zhuǎn)移情況。3應(yīng)用流式細(xì)胞術(shù)、血細(xì)胞計(jì)數(shù)儀檢測(cè)小鼠外周血中中性粒細(xì)胞情況,應(yīng)用免疫組織化學(xué)染色方法檢測(cè)小鼠肺組織中中性粒細(xì)胞浸潤(rùn)情況。4應(yīng)用脫氧核糖核酸酶(deoxyribonuclease,DNase-I)破壞中性粒細(xì)胞胞外捕獲網(wǎng)結(jié)構(gòu)(neutrophil extracellular trap,NET),肺組織病理切片觀察小鼠肺轉(zhuǎn)移情況。5統(tǒng)計(jì)學(xué)方法:應(yīng)用SAS9.1統(tǒng)計(jì)軟件包對(duì)數(shù)據(jù)進(jìn)行統(tǒng)計(jì)分析,所有計(jì)量資料用(均數(shù)±標(biāo)準(zhǔn)差)來(lái)表示。兩組樣本之間差異的顯著性分析采用t檢驗(yàn),多組樣本間差異的顯著性分析采用One-way ANOVA單因素方差分析,多組之間兩兩比較(事后檢驗(yàn))采用Bonferroni法。在炎性細(xì)胞浸潤(rùn)和組織破壞程度評(píng)估分析中,兩組樣本(多值有序變量的成組設(shè)計(jì)資料)之間采用Wilcoxon秩和檢驗(yàn)。P0.05視為有統(tǒng)計(jì)學(xué)意義。結(jié)果1 2.5%DSS可誘導(dǎo)Balb/c雌鼠明顯的結(jié)腸炎癥反應(yīng),實(shí)驗(yàn)過程中觀察可見經(jīng)DSS誘導(dǎo)的實(shí)驗(yàn)小鼠發(fā)生明顯腹瀉、血便,解剖見結(jié)腸組織明顯充血,長(zhǎng)度明顯縮短(P0.01),病理切片觀察結(jié)腸組織可見大量的炎性細(xì)胞浸潤(rùn)及組織損傷。2在DSS誘導(dǎo)的IBD乳腺癌肺轉(zhuǎn)移模型中,肺組織病理切片可見經(jīng)DSS誘導(dǎo)的小鼠,與正常飲水組相比,其乳腺癌肺轉(zhuǎn)移明顯增加(P0.01)。3血細(xì)胞計(jì)數(shù)、流式細(xì)胞術(shù)結(jié)果顯示,DSS可誘導(dǎo)小鼠外周血中中性粒細(xì)胞的動(dòng)員,其數(shù)量及比例均增加(P0.05),免疫組織化學(xué)染色結(jié)果顯示,轉(zhuǎn)移部位肺組織中浸潤(rùn)的中性粒細(xì)胞也明顯增加(P0.01)。4應(yīng)用DNase-I破壞中性粒細(xì)胞NET結(jié)構(gòu),可逆轉(zhuǎn)DSS誘導(dǎo)的IBD促進(jìn)乳腺癌肺轉(zhuǎn)移增加(P0.05)。結(jié)論DSS誘導(dǎo)的小鼠IBD乳腺癌肺轉(zhuǎn)移模型可見明顯的乳腺癌肺轉(zhuǎn)移,其機(jī)制可能為DSS誘導(dǎo)小鼠外周血中中性粒細(xì)胞動(dòng)員并浸潤(rùn)至肺組織,動(dòng)員的中性粒細(xì)胞可通過形成NET結(jié)構(gòu)促進(jìn)乳腺癌肺轉(zhuǎn)移,應(yīng)用脫氧核糖核酸酶(DNase-I)破壞中性粒細(xì)胞NET結(jié)構(gòu),可逆轉(zhuǎn)DSS誘導(dǎo)的小鼠乳腺癌肺轉(zhuǎn)移增加。
[Abstract]:Objective to investigate whether inflammatory bowel disease can accelerate the process of lung metastasis of breast cancer and explore its possible mechanism by establishing a model of lung metastasis in mice with inflammatory bowel disease. In order to provide a new intervention strategy for the prevention and treatment of distant metastasis in patients with breast cancer complicated with inflammatory bowel disease. Methods 1 the model of inflammatory bowel disease (inflammatory bowel disease,IBD) induced by 2.5% glucosulfate sodium sulfate (dextran sulfate sodium,DSS) was established in Balb/c female rats according to the experimental animal model. On the basis of this model, mice breast cancer cell line 4T1 100ul (1 脳 10 ~ 6 / ml) was injected into the tail vein to establish the animal model of lung metastasis of IBD breast cancer. The results of the animal experiment were repeated twice. At least 10 animals in each group were selected for each experiment (5 animals in each group were selected for statistical analysis). 2 colonic inflammation in IBD breast cancer lung metastasis model was evaluated by histopathological sections of colon. The lung metastasis in the model of IBD breast cancer was observed by pathological sections of lung tissue. 3The neutrophils in peripheral blood of mice were detected by flow cytometry and blood cell counter. Immunohistochemical staining was used to detect neutrophil infiltration in the lung tissue of mice. (4) the extracellular capture net structure (neutrophil extracellular trap,NET) of neutrophils was destroyed by deoxyribonuclease (deoxyribonuclease,DNase-I). The pathological sections of lung tissue were used to observe the lung metastasis in mice. 5 Statistical methods: the data were statistically analyzed by SAS9.1 software package, and all the data were expressed by the mean 鹵standard deviation (mean 鹵standard deviation). T-test was used to analyze the significance of the difference between the two groups, One-way ANOVA single factor analysis of variance was used to analyze the significance of the difference between multiple samples, and Bonferroni method was used to compare the differences between the two groups (post-test). In the evaluation analysis of inflammatory cell infiltration and tissue destruction, Wilcoxon rank sum test was used between the two groups of samples (group design data of multi-valued ordered variables). P0.05 was regarded as statistically significant. Results 1 2.5%DSS could induce obvious colonic inflammation in Balb/c female rats. In the course of the experiment, the mice induced by DSS had obvious diarrhea, blood stool, and the colon tissue was obviously hyperemia in anatomy, and in the course of the experiment, there were obvious diarrhea and blood stool in the experimental mice induced by DSS. The length was significantly shortened (P0.01), and a large number of inflammatory cell infiltration and tissue injury were observed in colonic tissue by pathological section. 2 in the lung metastasis model of IBD breast cancer induced by DSS, the mice induced by DSS could be seen in the pathological section of lung tissue. Compared with the normal drinking water group, the lung metastasis of breast cancer was significantly increased (P0.01). 3Blood cell count and flow cytometry showed that DSS could induce the mobilization of neutrophils in peripheral blood of mice, and the number and proportion of neutrophils were increased (P0.05). Immunohistochemical staining showed that the infiltration of neutrophils in the metastatic lung tissue was also significantly increased (P0.01). (4) the NET structure of neutrophils was destroyed by DNase-I. Reversible DSS-induced IBD enhanced lung metastasis in breast cancer (P0.05). Conclusion the lung metastasis of breast cancer in IBD mice induced by DSS may be related to the mobilization of neutrophils in peripheral blood and infiltration of neutrophils into lung tissue induced by DSS, and the mechanism may be that DSS can induce the mobilization of neutrophils in peripheral blood of mice. The mobilized neutrophils could promote the lung metastasis of breast cancer by forming NET structure and destroy the NET structure of neutrophils by using deoxyribonuclease (DNase-I) to reverse the increase of DSS-induced lung metastasis of breast cancer in mice.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R574;R737.9

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 關(guān)會(huì)會(huì);李彥珊;苗麗霞;袁海蓮;孫巖峰;王軍;趙揚(yáng);劉秋玲;;兒童腎母細(xì)胞瘤肺轉(zhuǎn)移臨床特點(diǎn)及其相關(guān)因素[J];武警醫(yī)學(xué);2017年11期

2 金利娜;吳鳴;楊玉雯;馬良坤;;卵巢黏液性囊腺癌空洞性肺轉(zhuǎn)移病例分析[J];中國(guó)實(shí)用婦科與產(chǎn)科雜志;2006年03期

3 馬游;隆凸性皮膚纖維肉瘤反復(fù)肺轉(zhuǎn)移1例[J];中國(guó)皮膚性病學(xué)雜志;1997年04期

4 艾文鈺;綜合治療原發(fā)性肝細(xì)胞肝癌伴肺轉(zhuǎn)移術(shù)后存活12年一例[J];桂林醫(yī)學(xué)院學(xué)報(bào);1995年03期

5 楊天恩;任傳富;潘慧泉;;微波熱療對(duì)小鼠Lewis腫瘤自發(fā)性肺轉(zhuǎn)移的影響和作用[J];天津醫(yī)藥;1987年04期

6 于開今;胃癌肺轉(zhuǎn)移誤為血行播散型肺結(jié)核1例[J];陜西醫(yī)學(xué)雜志;1988年06期

7 唐建武;張眾;謝豐培;曹輝;錢振超;;615系小鼠可移植性乳腺癌肺轉(zhuǎn)移的組織形態(tài)學(xué)觀察[J];大連醫(yī)學(xué)院學(xué)報(bào);1988年01期

8 曹雪濤;IFN-γ與TNF、IL-2聯(lián)合應(yīng)用能有效地阻止小鼠NK抵抗性腫瘤的肺轉(zhuǎn)移[J];國(guó)外醫(yī)學(xué)(免疫學(xué)分冊(cè));1989年02期

9 稅開森;許其瓊;;腎癌術(shù)后肺轉(zhuǎn)移晚期癌癥生存12年1例[J];昆明醫(yī)學(xué)院學(xué)報(bào);1989年04期

10 田培林;俞炎平;王美英;;大腸癌肺轉(zhuǎn)移X線診斷[J];浙江腫瘤通訊;1989年01期

相關(guān)會(huì)議論文 前10條

1 牟達(dá);匡安仁;馬曉娟;李素平;;~(131)I對(duì)甲癌肺轉(zhuǎn)移的循證療效[A];第四屆全國(guó)中青年核醫(yī)學(xué)學(xué)術(shù)會(huì)議論文匯編[C];2008年

2 楊宏宇;王宇帆;蘇文;李博文;楊輝俊;沈時(shí)岳;王鋒;林博;洪霞;;腺樣囊性癌伴肺轉(zhuǎn)移治療方案考量[A];2017全國(guó)口腔頜面——頭頸腫瘤外科學(xué)術(shù)研討會(huì)論文集[C];2017年

3 袁超;劉恒超;胡永全;陶健;李衛(wèi)鵬;;~(131)I治療分化型甲狀腺癌術(shù)后肺轉(zhuǎn)移的臨床研究[A];促進(jìn)科技經(jīng)濟(jì)結(jié)合，服務(wù)創(chuàng)新驅(qū)動(dòng)發(fā)展——蚌埠市科協(xié)2012年度學(xué)術(shù)年會(huì)論文集[C];2012年

4 李環(huán)宇;景士偉;;中子管中子對(duì)小鼠宮頸癌肺轉(zhuǎn)移治療作用的初步研究[A];中國(guó)核科學(xué)技術(shù)進(jìn)展報(bào)告（第四卷）——中國(guó)核學(xué)會(huì)2015年學(xué)術(shù)年會(huì)論文集第8冊(cè)（同位素分卷、輻射研究與應(yīng)用分卷、核技術(shù)工業(yè)應(yīng)用分卷、核農(nóng)學(xué)分卷、核醫(yī)學(xué)分卷、核情報(bào)分卷）[C];2015年

5 張媛;馮利;;乳腺癌骨轉(zhuǎn)移的術(shù)后輔助化學(xué)治療之中西進(jìn)展[A];2009年首屆全國(guó)中西醫(yī)腫瘤博士及中青年醫(yī)師論壇論文集[C];2009年

6 程晶;;乳腺癌骨轉(zhuǎn)移和骨相關(guān)疾病臨床診療專家共識(shí)(2010版)[A];湖北省第21屆腫瘤學(xué)術(shù)大會(huì)論文匯編[C];2011年

7 郭保鳳;劉勝;;乳腺癌骨轉(zhuǎn)移動(dòng)物模型的建立及中藥對(duì)其研究相關(guān)進(jìn)展[A];2011年中醫(yī)外科學(xué)術(shù)年會(huì)論文集[C];2011年

8 孫，

本文編號(hào)：2435680