【作者】 韩冰；

【导师】 刘国津； 范志民；

【作者基本信息】 吉林大学 ， 外科学， 2008， 博士

【摘要】 乳腺癌是危害女性生命和健康的最常见的恶性肿瘤,在我国约占全身恶性肿瘤的7%-10%,其发病率位于我国女性肿瘤的首位,且发病率逐年升高。乳腺癌的治疗方法也从外科手术治疗为主逐渐转变为个体化综合治疗的新时代。而在个体化综合治疗的各个组成部分中,乳腺癌的内分泌治疗起着非常重要的作用。随着对内分泌治疗的深入研究,人们的认识也逐渐从“内分泌机制”转向“细胞内分泌机制”。本实验采用免疫组化的方法,检测乳腺癌组织以及癌旁乳腺组织中催化雄激素和雌激素局部合成及分解的酶17β-HSD2和5的表达,并同时检测相应组织中CDC-47和其它类固醇激素受体(ERα、ERβ、PRA、PRB、AR)的表达。分别计算17β-HSD2和5两种酶和各种受体以及一些临床指标(例如:年龄,月经状态等),采用统计学方法分析它们之间差异以及相关性。结果发现17β-HSD 5在癌旁组织中阳性率明显高于癌组织中的阳性率,并且AR在癌旁组织与癌组织也有同样的差异,说明17β-HSD 5在乳腺癌的发生发展中起重要作用,也间接说明了雄激素通过AR而发挥了在乳腺组织中的保护作用。绝经后妇女17β-HSD 5表达明显高于绝经前妇女,这也反应绝经后妇女局部组织中雄激素代谢的改变。本实验证实了17β-HSD 5与孕激素代谢是有关的,在乳腺组织中17β-HSD 5通过其20aHSD活性降低局部孕激素水平。17β-HSD 2在乳腺癌患者的癌旁组织中高表达,在乳腺癌组织中则低表达,这种差异显示了17β-HSD 2在正常乳腺组织中具有保护作用。同时17β-HSD 2在癌旁组织中表达阳性率随肿瘤分期而降低,也证实了其在乳腺癌发生和发展中所具有的抑制作用。我们还发现ERα阳性的乳腺癌患者其肿瘤细胞分裂活性较低,比ERα阴性患者恶性度更低。通过本实验也为进一步研究做了准备。更多还原

【Abstract】 Breast cancer is the most common female malignant tumor which endangers the life and health of the female. It is about 7%-10% out of all malignant tumors in China. The incidence of the breast cancer is higher year by year. It is changing from the surgery to combined therapy along with we know more about the biology feature of the breast cancer. Now the therapy for the breast cancer includes operation, chemotherapy, radiotherapy, endocrine therapy, molecular targeted therapy and so on. With the using of the new medicine of endocrine therapy, the curative effect is more and more significant through the control of sex steroid level in both circulation and local tissue. And now the endocrine therapy for breast cancer has a more important role in the breast cancer combined therapy. Recently more and more scientists focus on the intracrinology of breast cancer, and the role of the local synthesis of the sex steroid in the endocrine target tissue becomes more important. Otherwise the function of the androgen in breast cancer is still controversial. Androgen therapy has been reported to have response rates of up to 46% in ER-positive patients. About 30% of breast cancer is ER or PR negative, and AR was found in about 49% of them. There is no effective endocrine therapy for them at present, so the using of androgen for them maybe become a method of endocrine therapy. Through the studying of the expression of 17β-hydroxysteroid dehydrogenase type 2 and type 5 in breast cancer and adjacent non-malignant tissue, and the correlations between the results and the clinical factors, we want to analyze the strength of the expression of 17β-HSD 2, 5 and the role of the two enzymes in the metabolism of sex steroid. We also analyze the relationship among the sex steroids by studying the correlation among the sex steroid receptors.Prepare the protein of type2 and type5 17β-HSD, injected sc with 1ml at multiple sites of New-Zealand rabbits. Collect the antiserum of the rabbits and purify it. Positive antiserum was analyzed by immunoblot. The expression of 17β- HSD2, 5 and CDC-47、ERα、ERβ、PRA、PRB、AR in 50 specimens of female human breast carcinoma and adjacent non-malignant tissues were studied by immunocytochemistry. These results were correlated with the clinical parameters.Results: 1、17β-HSD2 was found in the cytoplasm but not nucleus of tumor cells in 10% of cases and the stain was very weak. In adjacent non-malignant tissues, 92% of cases were positive and the enzyme was detected in the cytoplasm of epithelial cells of the acini and ducts. There was also no stain found in the nucleus, and the stain was mainly in the outer layer of the acini. The vessel wall was stained both in breast cancer tissue and adjacent non-malignant tissue. 17β-HSD5 was found in the cytoplasm of tumor cells in 44% of cases. In adjacent non-malignant tissues, 74% of cases were positive and the enzyme was detected in the cytoplasm of epithelial cells of the acini and ducts. No matter 17β-HSD2 or 17β-HSD5, there was no stain found in the nucleus and occasionally we found the stroma cell was positive. The control group was negative.There was a significant difference between the malignant tissue and the adjacent non-malignant tissues in the percentage of positive cells in both enzyme(x2=41.0000,P<.0001; x2=7.7586,P=0.0053). 2、All kinds of the receptors were found in the nuclei of tumor cells and epithelial cells of the acini and ducts except PRA and PRB. They also expressed in the cytoplasma and the stain was weak. The reason was not clear now. The percentage of the positive cells are 98%、74%、100%、42%、42%、84% in the malignant tissue and 48%、94%、100%、52%、36%、94% in the adjacent non-malignant tissues respectively. There was a significant difference between the malignant tissue and the adjacent non-malignant tissues in the percentage of positive cells in CDC-47 and ERα.(χ2= 25.0000, P<.0001;χ2= 8.3333, P=0.0039). 3、In malignant tissue, the expression of 17β-HSD2 was significantly correlated with the expression of 17β-HSD5 in adjacent non-malignant tissues (r=-0.29624,P=0.0367), the expression of PRA in adjacent non-malignant tissues(X2=5.1282,P=0.0235) and the menopause(X2=4.9257,P=0.0265). In adjacent non-malignant tissues, the expression of 17β-HSD2 was significantly correlated with the stage of the breast cancer (r=-0.28745 ,P=0.0430)and the expression of the CDC-47 in the adjacent non-malignant tissues(r=0.28873,P=0.0420). In adjacent non-malignant tissues, the expression of 17β-HSD5 was significantly correlated with the age of patients(r=0.34763,P=0.0134), menopause(r=0.39859,P=0.0046), the expression of ERβ(r=-0.31095,P=0.0280)in malignant tissue, the expression of PRA (X2=7.4871,P=0.0062)and ERα(X2=9.0835,P=0.0026) in adjacent non-malignant tissues. 4、In malignant tissue: the expression of AR was significantly correlated with the expression of ERα(r=0.28424,P=0.0454), the expression of PRA and PRB were significantly correlated with the expression of CDC-47(r=-0.42281,P=0.0025,r=-0.52952,P=<.0001), the expression of ERαwas significantly correlated with the expression of PRA , PRB and CDC-47(r=0.40318,P=0.0041; r=0.37860,P=0.0067; r=-0.46091,P=0.0008). In adjacent non-malignant tissues: the expression of PRA was significantly correlated with the age of patient(r=-0.31681,P=0.0282), the expression of ERβwas significantly correlated with the expression of CDC-47(r=0.39960,P=0.0044). No matter in malignant tissue or in adjacent non-malignant tissues the expression of PRA was significantly correlated with the expression of PRB (r=0.84010,P=<.0001,r=0.42457,P=0.0026). Conclusion: 1、In adjacent non-malignant tissues the expression of 17β-HSD5 was significantly higher than in cancer tissue, moreover AR was the same as 17β-HSD5. It indicated the important role of 17β-HSD5 in the development and/or progression of breast cancer, and it also suggested the androgen may suppress the breast cancer proliferation through AR.2、The expression of 17β-HSD5 of menopause women was significantly higher than premenopause ones in adjacent non-malignant tissues, this indicated the changing of the androgen local synthesized in the menopause women. 3、17β-HSD5 was able to regulate the metabolism of the progestogen by it’s 20a-HSD activity. 4、The significantly different expression of 17β-HSD 2 between the cancer tissue and the adjacent non-malignant tissues indicated it’s protective function in breast tissue. As the expression of 17β-HSD 2 in adjacent non-malignant tissues was significantly correlated with tumor stage, it indicated 17β-HSD 2 play an important role in the development and/or progression of breast cancer.5、because the tumor cells expressing ERαhad lower proliferative activity, ERαwas a good prognostic indicator.更多还原