| In veterinary clinics, canine breast tumor was a common tumor disease in female dogs.It shared many similarities with the breast tumor in humans, such as the etiology, the characteristics of tumor tissue, the therapeutic in medicine, the mammectomy and the prognosis.Therefore, canine of spontaneous breast tumor had become the most excellent models for human studying breast tumor disease.The pathohistology method、the immunohistochemistry techniques and molecular biology technology were employed in the experiment to carry out the investigations on the spontaneous canine breast tumor, in order to provide the diagnosises and the treatment basises to canine breast tumor.The purpose of this experiment is to study the expressions and significances of Claudin-1、E-cadherin、TNF-αand their relationship with clinical-pathologic parameters in canine breast tumor and to explore the correlation between Claudin-1、E-cadherin and TNF-a.1. The expression of Claudin-1 and E-cadherin in canine breast tumor.Claudins are tight junction proteins expressed by epithelial and endothelial cells. Cadherins are calcium-dependent cell-cell adhesion molecules that play critical roles during embryogenesis and in the maintenance of normal adult tissue architecture. In this experiment,10 cases of surgical operations excised doubtful canine breast tumor specimens were classified by the pathohistological observation.The immunohistochemistry method was used in the experiment which used two different kinds of rabbit anti-mouse polyclonal antibody to detect specimens and compare antibody expressions and pathology types. The results indicated that malignant breast tumor was to60%, in which mammary infiltrating ductal carcinoma was to 40%;benign tumor was to 20%, in which non-breast tumor 20%.The results showed that the rabbit anti-mouse polyclonal antibody Claudin-1 could react with the canine breast tumor tissues and the expression in benign breast cancer and non-breast tumor was significantly higher than breast tumor;The positive rate of Claudin-1 protein in malignant breast tumor is(2/6,33.3%)and that significantly lower than the normal breast(100.0%,2/2). Malignant breast tissue expression of E-cadherin-positive rate was 33.3%(2/6), significantly lower than normal mammary gland 100%(2/2).There is down-regulation of Claudin-1 and E-cadherin in breast carcinoma tissues.2. ELISA method was used to dectect the change of TNF-αlevel in serum.In this experiment, we used the method of ELISA to dectect TNF-αlevel in serum and erected the limit of health dogs (21.35±4.96 pg/ml).We choosed the female dogs of Labrador retriever, English spring spaniel and German Shepherd Dog.The results showed that breed couldn’t change the TNF-a levels of dogs’ serum. There is no statistically significant difference(P>0.05). But the risk of breast cancer in canine’ serum TNF-αwere significantly higher than that in healthy dogs and dogs with benign tumors.3. PCR method was used to analysis the expression of Claudin-1、E-cadherin and TNF-α.This study established the PCR to detect Claudin-1, E-cadherin, TNF-αgene expression. The method of molecular biology were used to detect normal breast and breast tumor tissues Claudin-1, E-cadherin, differences in gene expression, as well as serum TNF-αgene expression. The results sujested that this conclusion is same as the experimental land 2, Claudin-1, E-cadherin expression in normal breast was significantly higher than that in the expression of breast cancer, and in the malignant tumors expressed little, almost not expressed (Fig3-2,3-3). TNF-αin canine cancer is a high expression, in the normal tissue and benign tissues are also expressed (Fig3-4).It can provides a theoretical for clinical diagnosis and treatment from detecting Claudin-1, E-cadherin and TNF-α’s gene expression. |
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