小鼠喉粘膜和喉肿瘤的标记滞留细胞鉴定

发布时间：2018-03-19 03:01

本文选题：标记滞留细胞　切入点：干细胞　出处：《广州医学院》2011年硕士论文　论文类型：学位论文

【摘要】：【目的】:利用5-溴脱氧尿嘧啶核苷(BrdU)标记滞留细胞的方法研究小鼠喉粘膜和喉癌实体瘤内可能存在的干细胞及其分布特点。【方法】出生3天的昆明小鼠36只,随机分为实验组和对照组,每组18只。实验组小鼠皮下注射BrdU 50μg/g,对照组皮下注射生理盐水100μml,每天2次,共注射3d。于最后一次注射后2、4、8周分别处死6只小鼠,并取喉粘膜固定,石蜡包埋用于免疫组化检测,计算标记滞留细胞的阳性率。被BrdU标记的喉癌Hep细胞注入6只裸鼠皮下成瘤,8周后处死并取肿瘤固定,石蜡包埋用于HE染色及免疫组化检测。不同时间点标记滞留细胞阳性率的比较采用单因素方差分析,不同部位标记滞留细胞阳性率的比较采用最小显著差数法。【结果】实验组标记后2、4、8周,小鼠鳞状上皮和腺体上皮均有标记滞留细胞表达,对照组标记后无标记滞留细胞表达。不同时间点标记滞留细胞阳性率差异具有显著性(F=140.319,208.382 ;P=0.000,0.000)。鳞状上皮基底层中标记滞留细胞阳性率显著高于腺体细胞(t=6.376,10.944,15.376; P=0.000,0.000,0.000)。【结论】小鼠喉粘膜和喉癌实体瘤中存在标记滞留细胞,并随着观察时间的延长而逐渐减少,这些经过长时间观察而标记仍然存在的细胞被认为是喉粘膜成体干细胞及喉癌肿瘤干细胞。
[Abstract]:Objective: to study the possible existence and distribution of stem cells in mouse laryngeal mucosa and laryngeal carcinoma solid tumor by using 5-bromodeoxyuridine BrdU labeling method. [methods]. Thirty-six Kunming mice were randomly divided into experimental group (n = 18) and control group (n = 18). Mice in the experimental group were subcutaneously injected with BrdU 50 渭 g / g, and the control group were subcutaneously injected with 100 渭 ml of normal saline twice a day. Six mice were killed at 2 and 4 weeks after the last injection, and the laryngeal mucosa was fixed with paraffin embedded for immunohistochemical examination. BrdU labeled laryngeal carcinoma Hep cells were injected into 6 nude mice subcutaneously to form tumor cells for 8 weeks, and the tumor was fixed. Paraffin embedding was used for HE staining and immunohistochemical detection. The positive rate of labeled retained cells at different time points was compared by univariate ANOVA and the least significant difference method was used to compare the positive rates of labeled retained cells in different sites. [results]. In the experimental group, the labeled residual cells were expressed in the squamous epithelium and glandular epithelium of the mice at 2 and 4 weeks after labeling. In the control group, the positive rate of labeled retained cells was significantly different at different time points. The positive rate of labeled retained cells in the basal layer of squamous epithelium was significantly higher than that in the basal layer of squamous epithelium, and the positive rate of labeled cells in the basal layer of squamous epithelium was significantly higher than that in the glandular cells. [conclusion]. In mice laryngeal mucous membrane and laryngeal carcinoma solid tumor, the number of labeled residual cells decreased gradually with the prolongation of observation time. These cells, which have been observed for a long time and still exist, are believed to be laryngeal mucosal adult stem cells and laryngeal cancer tumor stem cells.
【学位授予单位】：广州医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.65

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