沙棘干乳剂对食管癌前病变的阻断作用和ANO1、EGFR在食管癌变进程中的表达及其意义

发布时间：2018-04-25 18:49

本文选题：食管癌 + 癌前病变　；参考：《山东大学》2017年硕士论文

【摘要】：目的建立化学致癌剂4-硝基喹啉-1-氧化物(4NQO)诱导的KM小鼠食管癌前病变模型;通过HE染色和病理组织学检测方法,观察并分析沙棘干乳剂对食管癌前病变的阻断作用。方法选取145只健康昆明小鼠,雌雄各半,随机分为4组:A组(空白对照组)10只、B组(单纯诱癌组)45只、C组(沙棘干乳剂治疗组)45只、D组(全反式维甲酸阳性对照组)45只。(1)A组:实验开始第1天～24周末,正常饮食、饮蒸馏水,不施加干预;B组:实验开始第1天～14周,自由饮用浓度0.1g/mL的4NQO水溶液,第14周末将4NQO水溶液改为饮用蒸馏水;C组:实验开始第1天～14周,自由饮用浓度O.1g/mL的4NQO水溶液,第14周末将4NQO水溶液改为饮用蒸馏水,并增加沙棘干乳剂(研究药物)灌胃,持续至实验结束;D组:实验开始第1天～14周,自由饮用浓度0.1g/mL的4NQO水溶液,第14周末将4NQO水溶液改为饮用蒸馏水,并增加全反式维甲酸(阳性对照药物)灌胃,持续至实验结束。(2)实验第10周末、第12周末、第14周末,均分别解剖A组小鼠2只、B组小鼠2只,于第14周末时,病理组织学检测结果证实模型建立成功;第19周末时,分别解剖A组小鼠2只,B、C、D组各组20只;24周末时,解剖各组剩余小鼠。通过HE染色和病理组织学检测方法纵向动态观察4NQO诱导的小鼠食管上皮黏膜组织发生癌前病变的变化过程;同时横向比较各组小鼠的食管上皮黏膜组织的增生程度,观察并分析沙棘干乳剂对小鼠食管癌前病变的阻断作用。结果(1)第14周末时:B组5例小鼠出现食管上皮黏膜组织轻度异型增生3例、中度异型增生1例、重度异型增生1例,异型增生率(轻+中+重度异型增生)为100%(5/5);(2)第19周末时:A、B、C、D四组同期相比,患癌率均无显著性差异(fisher P=0.927);B、C、D三组同期相比,轻+中度异型增生率、重度异型增生+癌变率均无显著性差异(χ~2=2.679,P0.05);(3)第24周末时:C组和B组同期相比,C组患癌率明显低于B组(χ~2=6.561,P0.05);D组与B组同期相比,D组患癌率明显低于B组(χ~2=10.506,P0.05);C组与D组同期相比,两组患癌率无显著性差异(χ~2=0.739,P0.05);结论(1)通过连续14周自由饮用浓度为0.1g/mL的4NQO水溶液的方法,可成功建立KM小鼠食管癌前病变模型;(2)沙棘干乳剂对食管癌前病变有一定的阻断作用,从而可以减缓食管癌前病变发展的进程,防止进一步恶变;且其阻断作用程度与全反式维甲酸相当。目的比较钙离子激活氯离子通道蛋白1(anoctamin1,ANO1)、表皮生长因子受体(Epidermal Growth Factor Receptor,EGFR)蛋白在小鼠不同食管上皮组织病变中的表达,探讨ANO1、EGFR蛋白表达与小鼠食管上皮黏膜组织病变的关系,为早期食管癌变诊断提供参考。方法应用化学致癌剂4硝基喹琳1氧化物(4NQO)由饮水法建立小鼠食管癌前病变模型,应用免疫组化方法检测小鼠不同食管上皮病变中EGFR、ANO1蛋白的表达水平。结果(1)ANO1表达定位于细胞浆,其中正常组织、轻度异型增生组织、中度异型增生组织、重度异型增生组织和癌变组织中的ANO1蛋白阳性表达率分别为 0%(0/14)、0%(0/25)、2.56%(1/39)、34.78%(8/23)和 50.00%(22/44);ANO1在癌组织中的阳性率显著高于正常组织(χ~2=11.278,P0.05)、轻度异型增生组织(χ~2=18.351,P0.05)及中度异型增生组织(χ~2=23.224,P0.05);ANO1在重度异型增生组织中的阳性率显著高于正常组织(fisher P=0.015)、轻度异型增生组织(χ~2=8.081,P0.05)及中度异型增生组织(χ~2=9.645,P0.05)。(2)EGFR表达定位于细胞浆,其中正常组织、轻度异型增生组织、中度异型增生组织、重度异型增生组织和癌变组织中的EGFR蛋白阳性表达率分别为7.14%(1/14)、16.00%(4/25)、23.08%(9/39)、52.17%(12/23)和 59.09%(26/44);EGFR在癌组织中的阳性率显著高于正常组织(χ~2=11.519,P0.05)、轻度异型增生组织(χ~2=12.046,P0.05)及中度异型增生组织(χ~2=10.996,P0.05);EGFR在重度异型增生组织中的阳性率显著高于正常组织(fisher P=0.011)、轻度异型增生组织(X2=7.054,P0.05)及中度异型增生组织(χ~2=5.469,P0.05)。67例重度异型增生及癌变组织中,ANO1表达阳性组中EGFR阳性率为93.33%(28/30),ANO1 表达阴性组中 EGFR 阴性率为 72.97%(27/37),ANO1 和 EGFR的表达相关系数r=0.665,相关性具有显著性意义(P0.05)。结论(1)ANO1在重度异型增生及癌变组织中表达特异性高,提示其参与食管癌的发生发展,并有可能成为新的食管癌药物治疗靶点和生物学标记;(2)ANO1、EGFR在食管重度异型增生及癌变组织中的表达呈一定正相关,两者的双向交互作用可能为其机制。
[Abstract]:Objective to establish a precancerous precancerous lesion model in KM mice induced by chemical carcinogen 4- nitroquinoline -1- oxide (4NQO), and to observe and analyze the blocking effect of seabuckthorn dry emulsion on precancerous lesions of the esophagus by HE staining and histopathological detection. Methods 145 healthy Kunming rats were selected and divided into 4 groups randomly: A group (blank control) Group B (group B) 45, group C (Seabuckthorn dry emulsion treatment group) 45, group D (all trans retinoic acid positive control group) 45. (1) group A: the experiment began first days to 24 weekend, normal diet, drink distilled water, no intervention; B group: the experiment began first to 14 weeks, free drinking concentration of 0.1g/mL 4NQO solution, fourteenth weekend 4NQO water solution, fourteenth weekend solution 4NQO water solution. The liquid was changed to distilled water for drinking. Group C: the experiment started first to 14 weeks, the free drinking concentration of 4NQO solution of O.1g/mL was free, the 4NQO solution was changed into drinking distilled water at the end of the fourteenth week, and the Seabuckthorn dry emulsion was added to the stomach and continued to the end of the experiment; group D: the experiment began first to 14 weeks, the free drinking concentration of 0.1g/mL 4NQO solution, fourteenth weeks At the end, the 4NQO water solution was changed into drinking distilled water, and all trans retinoic acid (positive control drugs) was added to the stomach and continued to the end of the experiment. (2) 2 mice in group A and 2 mice in group B were dissected respectively at the weekend of the experiment, twelfth weekend and fourteenth weekend. At the end of the fourteenth week, the pathological histological examination confirmed the success of the model; nineteenth weekend, respectively, respectively. 2 mice in group A, 20 rats in group B, C and D were dissected. At the 24 weekend, the remaining mice were dissected. The changes in the precancerous lesion of the esophageal epithelium induced by 4NQO were observed longitudinally by HE staining and histopathological detection, and the degree of proliferation of esophageal epithelial tissue in each group was compared and observed and observed. The blocking effect of seabuckthorn dry emulsion on precancerous lesions of the esophagus in mice was analyzed. Results (1) at the end of fourteenth weeks, 5 mice in group B showed mild dysplasia in esophageal epithelium, 1 cases of moderate dysplasia, 1 severe dysplasia, 100% (5/5) with the rate of dysplasia (mild + severe atypical growth), and (2) nineteenth weekend: A, B, C, and D four group of four phase There was no significant difference in the rate of cancer (Fisher P=0.927), and there was no significant difference in the rate of light + moderate dysplasia, severe dysplasia and cancer (P0.05) in the three groups of B, C and D. (3) at the twenty-fourth weekend, the cancer rate in the C and B groups was significantly lower than that in the B group (x ~2=6.561,). Compared with group B (x ~2=10.506, P0.05), there was no significant difference between the two groups in the C group and the D group (x ~2=0.739, P0.05), and (1) the esophageal precancerous lesion model of KM mice could be successfully established by the method of 4NQO aqueous solution of 0.1g/mL for 14 weeks of free drinking, and (2) the Seabuckthorn dry emulsion had a certain blocking effect on the precancerous lesions of the esophagus. It can be used to slow down the progression of precancerous lesions of the esophagus and prevent further malignant change; and its blocking effect is equivalent to all trans retinoic acid. Objective to compare calcium ion activated chloride channel protein 1 (anoctamin1, ANO1), Epidermal Growth Factor Receptor, EGFR protein in different esophageal epithelial tissues in mice The relationship between the expression of ANO1, EGFR protein and the pathological changes of esophageal epithelial mucosa in mice was discussed in order to provide reference for the diagnosis of early esophageal carcinogenesis. Methods the chemical carcinogen 4 nitroquinine 1 oxide (4NQO) was used to establish the precancerous precancerous lesion model of mice by drinking water method, and the different esophageal epithelial diseases in mice should be detected by immunohistochemical method. The expression level of EGFR, ANO1 protein. Results (1) the expression of ANO1 was located in the cytoplasm. The positive expression rates of ANO1 protein in normal tissue, mild dysplasia, severe dysplasia, severe dysplasia and cancerous tissues were 0% (0/14), 0% (0/25), 2.56% (1/39), 34.78% (8/23) and 50% (22/44); ANO1 in the cancer group. The positive rate in the fabric was significantly higher than that of normal tissue (x ~2=11.278, P0.05), mild dysplasia (x ~2=18.351, P0.05) and moderate dysplasia (x ~2=23.224, P0.05). The positive rate of ANO1 in severe dysplasia was significantly higher than that of normal tissue (Fisher P =0.015), and the mild dysplasia (chi ~2=8.081, P0.05) and moderate atypia were increased. (x ~2=9.645, P0.05). (2) the expression of EGFR was located in the cytoplasm. The positive rates of EGFR protein expression in normal tissue, mild dysplasia, severe dysplasia, severe dysplasia and cancerous tissues were 7.14% (1/14), 16% (4/ 25), 23.08% (9/39), 52.17% (12/23) and 59.09% (26/44); EGFR was in the cancer tissue. The positive rate was significantly higher than that of normal tissue (x ~2=11.519, P0.05), mild dysplasia (x ~2=12.046, P0.05) and moderate dysplasia (x ~2=10.996, P0.05). The positive rate of EGFR in severe dysplasia was significantly higher than that of normal tissue (Fisher P=0.011), mild dysplasia (X2=7.054, P0.05) and moderate dysplasia (P0.05). The positive rate of EGFR in ANO1 expression positive group was 93.33% (28/30), EGFR negative rate in ANO1 negative group was 72.97% (27/37) in ANO1 negative group, and the correlation coefficient of ANO1 and EGFR was significant (1) in severe dysplasia and cancerous tissue in.67 negative group.67. It has high expression specificity, suggesting that it participates in the development of esophageal cancer and may become a new target for the treatment of esophageal cancer and biological markers. (2) ANO1, EGFR is positively correlated with the expression of severe dysplasia and cancerous tissues of the esophagus, and the two-way interaction between them may be the mechanism.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.1

【相似文献】