4NQO饮水法诱发小鼠口腔癌淋巴道转移模型的改进以及骨髓播散细胞的提取及鉴定

发布时间：2018-05-05 21:01

本文选题：四硝基喹啉-1-氧化物 + 口腔癌　；参考：《广西医科大学》2013年硕士论文

【摘要】：研究目的：(1).通过比较四硝基喹啉-1-氧化物(4-nitroquinoline1-oxide,4-NQO)饮水法在不同给药时间诱发小鼠口腔癌淋巴道转移模型过程中口腔黏膜病变程度的差异,进一步改进小鼠口腔癌淋巴道转移模型。 (2).利用4-硝基喹啉-1-氧化物(4-nitroquinoline1-oxide,4-NQO)饮水法诱发小鼠口腔癌模型,提取不同时期的骨髓播散细胞,比较口腔黏膜癌变过程不同时期骨髓播散细胞的差异。研究方法：(1).70只balb/c小鼠,随机分为2组：实验组60只,给予200mg/L4NQ0饮水喂养8、14、20周(各20只),停药改用普通自来水喂养到45周；对照组10只小鼠,单纯饮用自来水。45周时通过肉眼和组织学观察各组其病变情况并比较小鼠体重的变化情况。 (2).通过第一部分完善构建动物模型的方法构建小鼠口腔癌变动物模型,选择病理检查明确诊断为正常舌粘膜的小鼠10例、舌单纯性增生的小鼠10例、舌轻中度异常增生的小鼠20例、舌重度异常增生的小鼠20例、舌高分化鳞癌的小鼠20例。收集所有小鼠的后腿股骨骨髓,用Ficoll密度梯度离心法提取单个核细胞并制成细胞涂片后,以PAN-CK上皮性特异性抗体为标记物,用免疫细胞化学SP法鉴定骨髓播散上皮细胞,比较不同时期的骨髓播散细胞的数目差异。结果：(1).随着给药时间的延长,实验组小鼠舌背黏膜相继出现白色斑块、溃疡、乳头状增生等改变,小鼠舌背产生轻度异常增生—中度异常增生—重度异常增生—原位癌—鳞癌的典型病理变化。给药8周组70.6%小鼠舌背黏膜表现为单纯性增生,29.4%为轻中重度异常增生,舌癌发生率为0%；给药14周组88.2%为轻中重度异常增生,11.8%为原位癌；给药20周组100%为鳞癌,并且100%转移颈部淋巴结；三组未见远处转移。14、20周组小鼠体重较给药前明显减轻(p0.05),8周组和对照组小鼠体重较给药前无明显变化(p0.05)。 (2).10例正常舌粘膜和10例单纯性增生,20例轻中度异常增生的小鼠骨髓细胞涂片均未能发现播散阳性细胞,而在20例舌重度异常增生的小鼠骨髓细胞涂片中有8例发现播散阳性细胞,占40%；20例鳞癌小鼠骨髓涂片中有13例发现播散阳性细胞,占65%。重度异常增生、鳞癌组两组之间播散细胞阳性率的比较有显著性差异(P0.05)。并发现骨髓播散细胞阳性组之间,随着小鼠舌黏膜逐步恶变,骨髓中的播散细胞数越多。结论：(1).在相同剂量下,随着给药时间延长,口腔黏膜癌前病变以及口腔癌发生率越高,但是体重减轻越明显。 (2).随着小鼠舌黏膜逐步恶变,在舌重度异常增生时,已经出现骨髓播散细胞,并且随着病变程度加重,骨髓播散细胞发生率和骨播散细胞数量增加。
[Abstract]:Objective: to study. In order to improve the lymphatic metastasis model of oral cancer in mice, the difference of oral mucosal lesion in mice induced by tetranitroquinoline 4-nitroquinoline 1-oxidetio (4-NQO) drinking water at different time was compared. There are two pieces of water. The mouse oral cancer model was induced by 4-nitroquinoline-1-oxide-4-NQO (4-nitroquinoline-1-oxide-4-NQO) drinking water. Bone marrow diffusing cells were extracted from different stages of oral mucosa carcinogenesis. The difference of bone marrow spreading cells in different stages of oral mucosa carcinogenesis was compared. Methods one hundred and seventy balb/c mice were randomly divided into two groups: the experimental group (n = 60) was fed with 200mg/L4NQ0 for 14 weeks (20 mice in each group) and the control group (n = 10) was fed with tap water for 45 weeks, and the control group (n = 10). After drinking tap water for 45 weeks, the pathological changes of each group were observed by naked eye and histology, and the changes of body weight of mice were compared. There are two pieces of water. In the first part, the animal model of oral carcinogenesis in mice was established by perfecting the animal model. Ten mice with normal tongue mucosa and 10 mice with simple hyperplasia of tongue were selected by pathological examination. There were 20 mice with mild to moderate dysplasia of tongue, 20 mice with severe dysplasia of tongue and 20 mice with highly differentiated squamous cell carcinoma of tongue. Bone marrow of femur of hind leg of all mice was collected. Mononuclear cells were extracted by Ficoll density gradient centrifugation and made into cell smears. Bone marrow disseminated epithelial cells were identified by immunocytochemistry SP method with PAN-CK epithelial-specific antibody as marker. To compare the number of bone marrow diffusing cells in different periods. The result is that one is one. With the prolongation of administration time, white plaques, ulcers, papillary hyperplasia and other changes appeared in the dorsal tongue mucosa of mice in the experimental group. The typical pathological changes of mouse tongue were mild dysplasia, moderate dysplasia, severe dysplasia, carcinoma in situ and squamous cell carcinoma. In the 8-week group, 70.6% of the mice showed simple hyperplasia of the tongue, 29.4% of the mice showed mild and severe dysplasia, and the incidence of tongue cancer was 0, 88.2% of the 14-week group showed mild and severe dysplasia in situ, and 100% of the 20 weeks of administration showed squamous cell carcinoma. The weight of the mice in the distant metastasis group was significantly lower than that in the control group and the control group at the 8th week after treatment, and there was no significant change in the body weight of the mice in the group of distant metastasis and in the control group compared with the control group before and after administration of the drug, and no significant change was found in the weight of the mice in the group of distant metastasis. Diffusion positive cells were not found in bone marrow cell smears of 10 cases of normal tongue mucosa and 10 cases of simple hyperplasia and 20 cases of mild to moderate dysplasia. Of the 20 mice with severe dysplasia of tongue, 8 cases were diffused positive cells in bone marrow smears, and 13 cases (65%) were positive cells in bone marrow smears of 20 cases of squamous cell carcinoma (SCC). There was a significant difference in the positive rate of disseminated cells between the two groups in severe dysplasia and squamous cell carcinoma group (P 0.05). It was also found that the number of disseminated cells in bone marrow increased with the gradual malignant change of tongue mucosa between the positive group of bone marrow diffusing cells. Conclusion: 1: 1. At the same dose, the oral precancerous lesion and oral cancer rate increased with the prolongation of administration time, but the weight loss was more obvious. There are two pieces of water. With the gradual malignant change of the mouse tongue mucosa, the bone marrow spreading cells appeared in the severe dysplasia of the tongue, and with the severity of the lesion, the incidence of bone marrow spreading cells and the number of bone dissemination cells increased.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R-332;R739.8

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