乳腺癌癌前病变大鼠模型的建立
|
摘要
【目的】:乳腺癌是一种严重危害妇女健康的恶性肿瘤,在西方发达国家已经成为妇女的第一位致死原因,上世纪末“乳腺癌多阶段发生模式”的提出,指出了正常乳腺上皮细胞向恶性转化经历了增生到非典型增生到原位癌再到浸润性癌的渐进过程。在发展成为浸润性癌之前,非典型增生和原位癌可能很长时间处于相对稳定状态,被称之为乳腺癌的癌前病变。它包括一部分不伴或伴有非典型增生的乳腺增生性疾病、导管内乳头状瘤、硬化性腺病、导管原位癌和小叶原位癌等。浸润癌癌前病变是一组在形态学和遗传学方面均有改变的疾病,是乳腺癌发生的必经阶段,而在癌变之前这一发展过程是可逆的。因此,乳腺癌前病变这一阶段的基础性研究和诊断治疗水平的提高,对乳腺癌的防治工作具有重要的预防及临床意义。
针对发病率迅速增高的趋势,为了控制乳腺癌的威胁,不仅需要合理的治疗策略,更需要有效的预防措施。目前欧美临床乳腺癌化学预防战略已对乳腺癌防治产生重大影响。对癌前病变的研究不仅可以探讨乳腺癌的发病机制,而且对其转归机制更具重要意义。鉴于乳腺癌的发生是一个长期的渐进性病理过程,存在取材困难等多种因素影响,难以在人类本身进行观察,因此建立适合的动物模型可以代替人类进行这一方面的研究,以提供良好的研究对象和材料,从而为进行癌前病变的转归机制探讨提供实验基础,对乳腺癌的预防有很大价值。
目前,建立乳腺增生和乳腺癌的动物模型比较成熟,遗憾的是乳腺癌癌前病变模型因难以把握实验终点而鲜有成功模型建立的报道,此模型建立的成功关键在于准确把握实验终点,若实验终点过晚,则大多数模型发展成为乳腺癌,若实验终点过早,则多数模型还处于增生阶段,致使我们无法对乳腺癌癌前病变进行观察和干预。
随着分子生物学研究的进展,目前已在乳腺良性增生性疾病或癌前疾病中发现了许多与乳腺癌进程相关的生物标志物,其结果表明在癌前病变期细胞增殖的调节性生物标记已经发生了明显变化。其中ER、PR、CerbB-2、Ki67、COX-2、TGF-β1等已被证明在乳腺细胞增殖过程中起重要调控作用,国内外研究已验证了其在乳腺导管非典型增生上皮细胞癌变过程中的作用及意义。乳腺某些癌前病变与浸润性癌具有同样的分子改变。通过对乳腺癌发展过程中乳腺组织ER、PR、CerbB-2、Ki67、COX-2、TGF-β1的表达,主要是在乳腺癌癌前病变组织中的表达情况,通过建立良好的乳腺癌癌前病变动物模型,可提供深入探讨发病原因和机制的研究基础,为实现提出效果好的预防措施提供可信、可行依据。
【方法】:30只SD大鼠随机分为4组,Ⅰ组:空白对照组6只;Ⅱ组:DMBA+雌孕激素序贯5天组8只;Ⅲ组:DMBA+雌孕激素序贯30天组8只;Ⅳ:DMBA单药组8只。60天后处死,记录大体形态变化、镜下病理改变和免疫组化ERα、ERβ、PR、CerbB-2、Ki67、COX-2、TGF-β1表达,观察并对照分析各组的状况。
【结果】:DMBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组的乳腺经病理诊断明确的癌前病变率分别为66.67%、42.86%、28.57%,DMBA+雌孕激素序贯5天组乳腺癌癌前病变发生率明显高于其余各组,与其他各组比较有统计学意义(P＜0.01)。免疫组化结果示:ERα、Ki67、COX-2、TGF-β1四项免疫组化指标,DNBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组与空白对照组比较有统计学意义(P＜0.01),阳性评分较高,DMBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组各组间比较无统计学意义(P＞0.05);免疫组化指标Erβ,DMBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组各组与空白对照组比较有统计学意义(P＜0.01),阳性评分较低,DMBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组各组间比较无统计学意义(P＞0.05);PR、CerbB-2两项免疫组化指标,空白对照组、DMBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组各组间比较均无统计学意义(P＞0.05)。
【结论】:DMBA+雌孕激素序贯5天组经病理诊断,乳腺癌癌前病变发生率明显高于其余各组;DMBA+雌孕激素序贯5天组经免疫组化检测,ERα、Ki67、COX-2、TGF-β1四项免疫组化指标均明显高于空白对照组,ERβ略低于空白对照组,ERα、ERβ、Ki67、COX-2、TGF-β1五项免疫组化指标与DMBA+雌孕激素序贯30天组和单纯DMBA组比较无明显差异。DMBA+雌孕激素序贯5天组建立乳腺癌癌前病变方法优于DMBA+雌孕激素序贯30天组和单纯DMBA组,是建立乳腺癌癌前病变模型较好的方法,简单易行,可以用于大批量建立乳腺癌癌前病变模型,为探讨乳腺癌的发生机制及干预方法建立良好的实验平台。
【Objective】:Breast cancer is a malignant tumor that endengering women's health seriously and have become the first cause of women's death in the Western developed countries.Putting forward "a multi-stage breast cancer model" on the end of the century,pointing out that the transformation from normal mammary epithelial cells to malignant experienced the gradual process of hyperplasia to atypical hyperplasia to carcinoma in situ to invasive carcinoma.In the development prior to becoming invasive carcinomas,atypical hyperplasia and carcinoma in situ may be a very long time at a relatively steady state,which is called precancerous lesions of breast cancer.It includes part without or with atypical hyperplasia hyperplastic breast disease,intraductal papilloma, sclerosing adenosis,ductal carcinoma in situ and lobular carcinoma in situand so on.Premalignant breast disease is a set of both morphological and genetic changes diseases,which must happened in stages and a prelude to malignant,and in the process of prior to malignant are reversible. Therefore,the basic research and the level of diagnosis and treatment of premalignant breast disease has a great significance of breast cancer prevention and control.
With the rapid increase in morbidity trends,in order to control the threat of breast cancer,the treatment requires not only a reasonable strategy,but also the need for effective preventive measures.Currently, Europe and the United States in the clinical strategy in breast cancer chemoprevention of breast cancer have a significant impact on prevention and treatment.Study of precancerous lesions can not only explore the pathogenesis of breast cancer,but also its mechanism is more the importance of vesting.In view of the occurrence of breast cancer is a long-term progressive pathological process,based on the difficulty of the existence of a number of factors,it is difficult to observe in the human itself,so the establishment of a suitable animal model can replace the human studies to carry out in order to provide a good research subjects and materials,so as to provide experimental basis for the prevention of premalignancy of breast cancer's transfer mechanism and have a great value.
At present,the establishment of breast hyperplasia and breast cancer animal model are mature,it is regrettable that model of precancerous lesions are little to success for difficult to determine the end of experiment,if the end of the experiment is too late,then the majority of models developed into breast cancer,if the premature end of the experiment,the majority of models are still in the proliferative stage, resulting in a lower success rate.
With the progress of the study of molecular biology,a number of process-related biomarkers has been found in benign breast disease or precancerous diseases,the results show that the regulation of cell proliferation biomarkers have already significant changed in the period of precancerous lesions.In which ER,PR,CerbB-2,Ki67,COX-2,TGF-β1 have been proved playing an important role in the course of regulation of breast cell proliferation and has been validated have a great significance in the process of epithelial cells Carcinogenesis both at home and abroad.Some precancerous lesions of breast have the same molecular changes with invasive carcinomas.Establishment animal models of premalignant breast disease and research on ER,PR,CerbB-2,Ki67, COX-2,TGF-β1 in premalignant breast disease,may provide a research base which could explore in depth the causes and mechanism of pathogenesis, To provide credible and viable basis for achieving the preventive effect.
【Methods】:30 SD ratswere randomly divided into 4 groups,GroupⅠ:blank control 6;GroupⅡ:DMBA +hormone sequential 5 days 8;GroupⅢ:DMBA +hormone sequential 30 days 8;GroupⅣ:DMBA only 8.The general morphology、pathology and immunohistochemieal expression of ER、PR、CerbB-2、Ki67、COX-2,TGF-β1 were observed When the rats were executed after 60 days.
【Results】:The incidence of premalignant breast disease in groupⅡ(66.67%) was significantly higher than groupⅠ(0%)、groupⅢ(42.86%) and groupⅣ(28.57%)(P＜0.01).Immunohistochemistry results showed that the expression of ERα、Ki67、COX-2、TGF-β1 in groupⅡ,groupⅢand groupⅣhas statistical significance to groupⅠ(P＜0.01) and have a higher positive score,groupⅡ、groupⅢand groupⅣhas not statistical significance(P＞0.05);Erβin groupⅡ,groupⅢand groupⅣhas statistical significance to groupⅠ(P＜0.01) and have a low positive score,groupⅡ、groupⅢand groupⅣhas not statistical significance (P＞0.05).PR、CerbB-2 in groupⅠ、groupⅡ、groupⅢand groupⅣcomparison of each group were not statistically significant(P＞0.05).
【Conclusion:】:The result suggest DMBA+hormone sequential 5 days method is the best way to establish rat models of breast disease;The incidence of ERα,Ki67,COX-2,TGF-β1 in premalignant breast disease were significantly higher than the groupⅠ,ERβwas slightly lower than the groupⅠ,ERα,ERβ,Ki67,COX-2,TGFβ1 immunohistochemistry five indicators in groupⅡhas the roughly same with the groupⅢand groupⅣ.PR,CerbB-2,in premalignant breast disease has not changed significantly and to be further explored.
针对发病率迅速增高的趋势,为了控制乳腺癌的威胁,不仅需要合理的治疗策略,更需要有效的预防措施。目前欧美临床乳腺癌化学预防战略已对乳腺癌防治产生重大影响。对癌前病变的研究不仅可以探讨乳腺癌的发病机制,而且对其转归机制更具重要意义。鉴于乳腺癌的发生是一个长期的渐进性病理过程,存在取材困难等多种因素影响,难以在人类本身进行观察,因此建立适合的动物模型可以代替人类进行这一方面的研究,以提供良好的研究对象和材料,从而为进行癌前病变的转归机制探讨提供实验基础,对乳腺癌的预防有很大价值。
目前,建立乳腺增生和乳腺癌的动物模型比较成熟,遗憾的是乳腺癌癌前病变模型因难以把握实验终点而鲜有成功模型建立的报道,此模型建立的成功关键在于准确把握实验终点,若实验终点过晚,则大多数模型发展成为乳腺癌,若实验终点过早,则多数模型还处于增生阶段,致使我们无法对乳腺癌癌前病变进行观察和干预。
随着分子生物学研究的进展,目前已在乳腺良性增生性疾病或癌前疾病中发现了许多与乳腺癌进程相关的生物标志物,其结果表明在癌前病变期细胞增殖的调节性生物标记已经发生了明显变化。其中ER、PR、CerbB-2、Ki67、COX-2、TGF-β1等已被证明在乳腺细胞增殖过程中起重要调控作用,国内外研究已验证了其在乳腺导管非典型增生上皮细胞癌变过程中的作用及意义。乳腺某些癌前病变与浸润性癌具有同样的分子改变。通过对乳腺癌发展过程中乳腺组织ER、PR、CerbB-2、Ki67、COX-2、TGF-β1的表达,主要是在乳腺癌癌前病变组织中的表达情况,通过建立良好的乳腺癌癌前病变动物模型,可提供深入探讨发病原因和机制的研究基础,为实现提出效果好的预防措施提供可信、可行依据。
【方法】:30只SD大鼠随机分为4组,Ⅰ组:空白对照组6只;Ⅱ组:DMBA+雌孕激素序贯5天组8只;Ⅲ组:DMBA+雌孕激素序贯30天组8只;Ⅳ:DMBA单药组8只。60天后处死,记录大体形态变化、镜下病理改变和免疫组化ERα、ERβ、PR、CerbB-2、Ki67、COX-2、TGF-β1表达,观察并对照分析各组的状况。
【结果】:DMBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组的乳腺经病理诊断明确的癌前病变率分别为66.67%、42.86%、28.57%,DMBA+雌孕激素序贯5天组乳腺癌癌前病变发生率明显高于其余各组,与其他各组比较有统计学意义(P＜0.01)。免疫组化结果示:ERα、Ki67、COX-2、TGF-β1四项免疫组化指标,DNBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组与空白对照组比较有统计学意义(P＜0.01),阳性评分较高,DMBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组各组间比较无统计学意义(P＞0.05);免疫组化指标Erβ,DMBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组各组与空白对照组比较有统计学意义(P＜0.01),阳性评分较低,DMBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组各组间比较无统计学意义(P＞0.05);PR、CerbB-2两项免疫组化指标,空白对照组、DMBA+雌孕激素序贯5天组、DMBA+雌孕激素序贯30天组、DMBA单药组各组间比较均无统计学意义(P＞0.05)。
【结论】:DMBA+雌孕激素序贯5天组经病理诊断,乳腺癌癌前病变发生率明显高于其余各组;DMBA+雌孕激素序贯5天组经免疫组化检测,ERα、Ki67、COX-2、TGF-β1四项免疫组化指标均明显高于空白对照组,ERβ略低于空白对照组,ERα、ERβ、Ki67、COX-2、TGF-β1五项免疫组化指标与DMBA+雌孕激素序贯30天组和单纯DMBA组比较无明显差异。DMBA+雌孕激素序贯5天组建立乳腺癌癌前病变方法优于DMBA+雌孕激素序贯30天组和单纯DMBA组,是建立乳腺癌癌前病变模型较好的方法,简单易行,可以用于大批量建立乳腺癌癌前病变模型,为探讨乳腺癌的发生机制及干预方法建立良好的实验平台。
【Objective】:Breast cancer is a malignant tumor that endengering women's health seriously and have become the first cause of women's death in the Western developed countries.Putting forward "a multi-stage breast cancer model" on the end of the century,pointing out that the transformation from normal mammary epithelial cells to malignant experienced the gradual process of hyperplasia to atypical hyperplasia to carcinoma in situ to invasive carcinoma.In the development prior to becoming invasive carcinomas,atypical hyperplasia and carcinoma in situ may be a very long time at a relatively steady state,which is called precancerous lesions of breast cancer.It includes part without or with atypical hyperplasia hyperplastic breast disease,intraductal papilloma, sclerosing adenosis,ductal carcinoma in situ and lobular carcinoma in situand so on.Premalignant breast disease is a set of both morphological and genetic changes diseases,which must happened in stages and a prelude to malignant,and in the process of prior to malignant are reversible. Therefore,the basic research and the level of diagnosis and treatment of premalignant breast disease has a great significance of breast cancer prevention and control.
With the rapid increase in morbidity trends,in order to control the threat of breast cancer,the treatment requires not only a reasonable strategy,but also the need for effective preventive measures.Currently, Europe and the United States in the clinical strategy in breast cancer chemoprevention of breast cancer have a significant impact on prevention and treatment.Study of precancerous lesions can not only explore the pathogenesis of breast cancer,but also its mechanism is more the importance of vesting.In view of the occurrence of breast cancer is a long-term progressive pathological process,based on the difficulty of the existence of a number of factors,it is difficult to observe in the human itself,so the establishment of a suitable animal model can replace the human studies to carry out in order to provide a good research subjects and materials,so as to provide experimental basis for the prevention of premalignancy of breast cancer's transfer mechanism and have a great value.
At present,the establishment of breast hyperplasia and breast cancer animal model are mature,it is regrettable that model of precancerous lesions are little to success for difficult to determine the end of experiment,if the end of the experiment is too late,then the majority of models developed into breast cancer,if the premature end of the experiment,the majority of models are still in the proliferative stage, resulting in a lower success rate.
With the progress of the study of molecular biology,a number of process-related biomarkers has been found in benign breast disease or precancerous diseases,the results show that the regulation of cell proliferation biomarkers have already significant changed in the period of precancerous lesions.In which ER,PR,CerbB-2,Ki67,COX-2,TGF-β1 have been proved playing an important role in the course of regulation of breast cell proliferation and has been validated have a great significance in the process of epithelial cells Carcinogenesis both at home and abroad.Some precancerous lesions of breast have the same molecular changes with invasive carcinomas.Establishment animal models of premalignant breast disease and research on ER,PR,CerbB-2,Ki67, COX-2,TGF-β1 in premalignant breast disease,may provide a research base which could explore in depth the causes and mechanism of pathogenesis, To provide credible and viable basis for achieving the preventive effect.
【Methods】:30 SD ratswere randomly divided into 4 groups,GroupⅠ:blank control 6;GroupⅡ:DMBA +hormone sequential 5 days 8;GroupⅢ:DMBA +hormone sequential 30 days 8;GroupⅣ:DMBA only 8.The general morphology、pathology and immunohistochemieal expression of ER、PR、CerbB-2、Ki67、COX-2,TGF-β1 were observed When the rats were executed after 60 days.
【Results】:The incidence of premalignant breast disease in groupⅡ(66.67%) was significantly higher than groupⅠ(0%)、groupⅢ(42.86%) and groupⅣ(28.57%)(P＜0.01).Immunohistochemistry results showed that the expression of ERα、Ki67、COX-2、TGF-β1 in groupⅡ,groupⅢand groupⅣhas statistical significance to groupⅠ(P＜0.01) and have a higher positive score,groupⅡ、groupⅢand groupⅣhas not statistical significance(P＞0.05);Erβin groupⅡ,groupⅢand groupⅣhas statistical significance to groupⅠ(P＜0.01) and have a low positive score,groupⅡ、groupⅢand groupⅣhas not statistical significance (P＞0.05).PR、CerbB-2 in groupⅠ、groupⅡ、groupⅢand groupⅣcomparison of each group were not statistically significant(P＞0.05).
【Conclusion:】:The result suggest DMBA+hormone sequential 5 days method is the best way to establish rat models of breast disease;The incidence of ERα,Ki67,COX-2,TGF-β1 in premalignant breast disease were significantly higher than the groupⅠ,ERβwas slightly lower than the groupⅠ,ERα,ERβ,Ki67,COX-2,TGFβ1 immunohistochemistry five indicators in groupⅡhas the roughly same with the groupⅢand groupⅣ.PR,CerbB-2,in premalignant breast disease has not changed significantly and to be further explored.
引文
[1].Rockhill B , Spiegelman D, Byrne C, et al. Vaidation of the Gail Model of breast cancer risk prediction and implications for chemprevetion.J Natl Cancer Inst, 2001, 93(5): 358-366
[2].Stute P, Szuwart T, Schlueter M, et al. Effects of hormone therapy on estrogen synthesis from E1S in the mammary gland of postmenopausal women. Maturitas, 2008, 59(2): 163-173
[3].Di Tommaso L, Franchi G, Destro A, et al. Toker cells of the breast. Morphological and immunohistochemical characterization of 40 cases. Hum Pathol, 2008, 39(9): 1295-1300
[4]. Kim JH, Na HK, Pak YK, et al. Roles of ERK and p38 mitogen-activated protein kinases in phorbol ester-induced NF-kappaB activation and COX-2 expression in human breast epithelial cells. Chem Biol Interact, 2008, 171(2): 133-141
[5]. Calaf GM, Echiburu-Chau C, Zhao YL, et al. BigH3 protein expression as a marker for breast cancer. Int J Mol Med, 2008, 21 (5): 561-568
[6]. Yang WT, Lewis MT, Hess K, et al. Decreased TGFβ signaling and increased C0X2 expression in high risk women with increased mammographic breast density. Breast Cancer Res Treat, 2009, [Epub ahead of print] [7].O' ConnellP, PekkelV, FuquaSA, etal. Analysis of loss of heterozygosity in 399 premalignant breast lesions at 15 genetic loci. J NSTL Cancer inst, 1998, 90(16): 697- 730
[8]. BaiM, Agnantis NJ, kamin S, etal. In vivo cell kinetics in breast carcinogenesis. Breast Cancer Res, 2001, 3(4): 276-283
[9]. Jose Russo, Irma H. Russo. Atlas and histologic Classification of tumors of the rat mammary gland. Mammary Gland Biology and Neoplasia, 2000, 5(2): 187-200
[10]. Hartmann L C, Schaid D J, Woods J E, et al. Efficacy of bilateral prophylactic mastectomy in women with a family history of breast cancer. N Engl Med,1999,340:77-84
[11].Herschkowitz JI,Simin K,WeigmanVJ,et al.Identification of conserved gene expression features between murine mammary carcinomamodels and human breast tumors.Genome Biol,2007,8(5):76
[12].李静蔚,宋爱莉,张敬涛等.DMBA诱导大鼠乳腺癌癌前病变的组织病理学研究.中华中医药学刊,2008,(3):26
[13].Henry J.Thompson,John N.McGinley,Pamela Wolfe,et al.Temporal sequence of mammary intraductal proliferations,ductal carcinomas in situ and adenocarcinomas induced by 1-methy-1-nitrosourea in rats.Carcinogenesis vol,1998,19(12):21 81-2185
[14].Sue C Heffelfinger,Mei Yan,Robin B Gear,et al.Inhibition of VEGFR2 prevents DMBA-induced mammary tumor formation.Laboratory nvestigation,2004,84:989-998
[15].P.M.Gullino,H.M.Pettigrew,F.H.Grantham.N-nitrosomethylurea as mammary gland carcinogen in rats.Natl.Cancer Inst,1975,54:401-414
[16].Colditz GA,Hankinson SE,Hunter DJ,et al.The use of estrogens and progestins and the risk of breast cancer in postmenopausal women.N Engl J Med,1995,332:1589-1593
[17].V K Turan,R I Sanchez,J J Li,et al.The effects of steroidal estrogens in ACI rat mammary carcinogenesis:17β-estradiol,2-hydroxyestradiol,4-hydroxyestr- adiol,16α-hydroxyestradiol,and 4-hydroxyestrone.Journal of Endocrinology,2004,183:91-99
[18].余望贻,姜海斌,孟琼等.乳腺增生病动物模型的实验研究.中国实验动物学报,2004,(4):12
[19].Harvey JM,Clark GM,Osborne CK,et al.Estrogen receptor status by immunohistochemistry is superior to the lig-and-binding assay for predicting response to adjuvant endocrine therapy in breast cance.J Clin Oncol,1999,17(5):1474-1481
[20].Lawson JS,Field AS,Champion S,et al.Low oestrogen receptor alpha expression in normal breast tissue underlieslow breast cancer incidence in Japan.Lancet,1999,354(9192):1787-1788
[21].Allred DC,Mohsin SK,Fuqua SA.Histological and biological evolution of human premalignant breast disease.Endocr Relat Cancer,2001,8(1):47-61
[22].Shoker BS,Jarvis C,Sibson DR,et al.Oestrogen receptorexpression in the normal and pre-cancerous breast.JPathol,1999,188(3):237-244
[23].Skliris GP,Munot K,Bell SM,et al.Reduced expression of oestrogen using DNA methyl transferase inhibitors in a cell line model.J Pathol,2003,201(2):213-220
[24].Shaban AM,Neill PA,Davies MP,et al.Declining estrogen receptorexpression defines malignant progression of human breast neoplasia[J].Am J Surg Pathol,2003,27(12);1502-1512
[25].Anderson E.The role of oestrogen and progesterone recep-tors in human mammary development and tumorigenesis.Breast Cancer Res,2002,4(5):197-201
[26].Quenel N,Wafflart J,Bergonie F.The prognostic value of CerbB-2 in primary breast carcinoma.A study on 942 cases.Breast Cancer Research and Treatment,1995,3:283-285
[27].浦勇顺,周金良,王芳等.p53和C-erbB-2过表达与乳腺癌预后的关系.上海第二医科大学学报,2001,21(2):170
[28].LiBJ,Zhu ZH,Wang JY.Expression correlation of Ki67 to P53,VEGF,and cerbB-2 genes in breast cancer and their clinicalsignifican.Ai Zheng,2004,23(10):1176-1179
[29].Bhatavdekar JM,PatelDD,ShalNG.Prognostic significance of immunohisto- chemically localized biomarkers in stage Ⅱ and stage Ⅲbreast cancer:a multivariate.Ann Surgoncol,2000,7(4):305-311
[30].RangerGS,JewellA,ThomasV,et al.Elevated expression of COX-2 in breast cancer and ductal carcinoma in situ has no correlation with established prognostic markers. J Surg Onco, 2004, 88(2): 100-103
[31]. Nakopoulou L, Mylona E, Papadaki I, et a. Overexpression of cyclooxygenase-2 is associated with a favorable prognostic phenotype in breast carcinoma. Pathobiol- ogy, 2005, 72(5): 241-249
[32]. Leivonen S K, Kahari V M. Transforming growth factor-beta signaling in cancer invasion and metastasis. Int J Cancer, 2007 , 121 (10) : 2119-2124
[33]. Desruisseau S, Palmari J, Giusti C, et al. Determination of TGF-betal protein level in human primary breast cancers and its relationship with survival. Br J Cancer, 2006, 94(2): 239-246
[1].Gullino PM,Pettigrew HM,Grantham FH.N-nitrosomethylureaas mammary gland carcinogen in rats.J Natl Cancer Inst.,1975,54:401-414
[2].Welsch CW.Host factors affecting the growth of carcinogen-induced rat mammary carcinomas:A review and tribute to Charles Brenton Huggins.Cancer Res,1985,45:3415-3443
[3].Harvell DME,Strecker TE,Tochacek M,et al.Rat strainspecic actions of 17b-estradiol in the mammary gland:Correlation between estrogen-induced lobuloalveolar hyperplasia and susceptibility to estrogen-induced mammary cancers.Proc Natl Acad Sci USA,2000,97:2779-2784
[4].Li SA,Liao JD,Li JJ.Estrogen-induced breast cancer in female ACI rats.New York:Springer-Verlag,2000,20:178-188
[5].Colerangle JB,RoyD.Perturbation of cell cycle kinetics in the mammary gland by stilbene estrogen,diethylstilbestrol.Cancer Lett,1995,94:55-63
[6].Xie B,Tsao SW,Wong YC.Induction of high incidence of mammary tumor in female Noble rats with a combination of 17b-oestradiol and testosterone.Carcinogenesis 1999,20:1069-1078
[7].P.M.Gullino,H.M.Pettigrew,F.H.Grantham.N-nitrosomethylureaas mammary gland carcinogen in rats.J.Natl.Cancer Inst,1975,54:401-414
[8].饶金才,李兰珍,陈云生等.乳腺增生病动物模型的复制及病理类型.中国病理生理杂志,1992,(6):8
[9].甘长清,吴凯南,苏新良等.三苯氧胺对大鼠乳腺增生形态学的影响.重庆医科大学学报,2003,(4):28
[10].吴建新,陆德铭,李道坊等.“乳块消”治疗乳腺增生病的临床及实验研究.上海中医学院,上海市中医药研究院学报,1989,(1):17
[11].余望贻,姜海斌,孟琼等.乳腺增生病动物模型的实验研究.中国实验动物学报,2004,(4):12
[12].吴在德.外科学.第5版.北京:人民卫生出版社,2002,355
[13].姚泰.生理学.第5版.北京:人民卫生出版社,2000,12-413
[14].Djuana M.E.Harvell,Tracy K Strecker,Martin Tochacek,et al.Rat strain-specific actions of 17β-estradiol in the mammary gland:Correlation between estrogen-induced lobuloalveolar hyperplasia and susceptibility to estrogen-induced mammary cancers.Medical Sciences,2000,97(6):2779-2784
[15].Irma H.Russo,Jose Russo.Mammary Gland Neoplasia in Long-Term Rodent Studies.Environmental Health Perspectives,1996,104(9):938-967
[16].Welsch CW,Medina D,KidwellW,et al.Rodent models to examine invivo hormonal regulation of mammary gland tumorigenesis.NewYork.Plenum Press,1987,63-179
[17].Callahan R,Neville MC,DanielCW,et al.Retrovirus and proto-oncogene involvement in the etiology of breast neoplasia.New York:Plenum Press,1987,323-351
[18].Teich N,Wyke J,Mak T,et al.Pathogenesis of retrovirus-induced disease.NY:Cold Spring Harbor Laboratory,1982,20:785-998
[19].MacKenzie WF,Garner FM.Comparison of neoplasms in six sources of rats.J Nad CancerInst,1973,(50):1234-1257
[20].Haseman JF,HuffJ,Boorman GA.Use of historicalcontrol data in carcinogenicity studiesin rodents.Toxicol Pathol,1984,(12):126-135
[21].Ross MH,Bras G.Lasting influence of earlycaloric restriction on prevalence of neoplasms in the rat.J Natl Cancer Inst,1971,(47):1095-1113
[22].Tarone RE,Chu KC,Ward JM.Variability inthe rates of some common naturally occurringtumors in Fischer 344 rats and(C57BL/6N x C3H/HeN)F1(B6C3F1) mice.J Natl CancerInst,1981,(66):1175-1181
[23].杨光伦,姚棒祥.外源性雌激素及三苯氧胺对大鼠乳腺癌癌前期病变的作用.中华普通外科杂志,2001,(2):16
[24].Russo J,Gusterson BA,Rogers AE,et al.Comparativestudy of human and rat mammary tumorigenesis.Lab Invest,1990,(62):1-32
[25].Henry J.Thompsonl,John McGinley,Kim Rothhammer,et al.Ovarian hormone dependence of pre-malignant and malignant mammary gland lesions induced in pre-pubertal rats by 1-methyl-1-nitrosourea.Carcinogenesis,1998,19(3):383-386.
[26].Russo J,Gusterson BA,RogersAE,et al.Comparative study of human and rat mammary tumorigenesis.Lab Invest,1990,62(3):244-278
[27].S A Li,S J Weroha,O Tawfikl,et al.Prevention of solely estrogen-induced mammary tumors in female ACI rats by tamoxifen:evidence for estrogen receptor mediation.J Endocrinol,2002,175(2):297-305
[28].Sue C Heffelfinger,Mei Yan,Robin B Gear,et al.Inhibition of VEGFR2prevents DMBA-induced mammary tumor formation.Laboratory Investigation,2004,(84):989-998
[29].李静蔚,宋爱莉,张维东等.乳腺非典型增生病与血管生成关系的实验研究.肿瘤防治杂志,2005,(12):13
[30].Dawson PJ,Wolman SR,Tait L,et al.MCFIOAT:a model for the evolution of cancer from proliferative breast disease.Am J Pathol,1996,148(1):313-319
[31].Miller FR.Xenograft Models of Premalignant Breast Disease.J Mammary Gland Biol Neoplasia,2000,5(4):379-391
[32].HeppnerGH,Wolman SR.MCF—lOAT:A model for human breast cancer development.Breast J,1999,5(2):122-129
[33].Shull JD,Spady TJ,Snyder MC,et al.Ovary-intact,but not ovariectomized female ACI rats treated with 17-estradiol rapidly develop mammary carcinoma.Carcinogenesis,1997,18:1595-1601
[34].傅西林,李树玲,范宇等.乳腺癌癌前病变与乳腺癌相关的多指标病理学研究.中国肿瘤临床,1999,(1):26
[35].Henry J.Thompsonl,3 and meenakshi sing h2 rat models of premalignant breast disease.J.Cancer,2006,20:985-994
[2].Stute P, Szuwart T, Schlueter M, et al. Effects of hormone therapy on estrogen synthesis from E1S in the mammary gland of postmenopausal women. Maturitas, 2008, 59(2): 163-173
[3].Di Tommaso L, Franchi G, Destro A, et al. Toker cells of the breast. Morphological and immunohistochemical characterization of 40 cases. Hum Pathol, 2008, 39(9): 1295-1300
[4]. Kim JH, Na HK, Pak YK, et al. Roles of ERK and p38 mitogen-activated protein kinases in phorbol ester-induced NF-kappaB activation and COX-2 expression in human breast epithelial cells. Chem Biol Interact, 2008, 171(2): 133-141
[5]. Calaf GM, Echiburu-Chau C, Zhao YL, et al. BigH3 protein expression as a marker for breast cancer. Int J Mol Med, 2008, 21 (5): 561-568
[6]. Yang WT, Lewis MT, Hess K, et al. Decreased TGFβ signaling and increased C0X2 expression in high risk women with increased mammographic breast density. Breast Cancer Res Treat, 2009, [Epub ahead of print] [7].O' ConnellP, PekkelV, FuquaSA, etal. Analysis of loss of heterozygosity in 399 premalignant breast lesions at 15 genetic loci. J NSTL Cancer inst, 1998, 90(16): 697- 730
[8]. BaiM, Agnantis NJ, kamin S, etal. In vivo cell kinetics in breast carcinogenesis. Breast Cancer Res, 2001, 3(4): 276-283
[9]. Jose Russo, Irma H. Russo. Atlas and histologic Classification of tumors of the rat mammary gland. Mammary Gland Biology and Neoplasia, 2000, 5(2): 187-200
[10]. Hartmann L C, Schaid D J, Woods J E, et al. Efficacy of bilateral prophylactic mastectomy in women with a family history of breast cancer. N Engl Med,1999,340:77-84
[11].Herschkowitz JI,Simin K,WeigmanVJ,et al.Identification of conserved gene expression features between murine mammary carcinomamodels and human breast tumors.Genome Biol,2007,8(5):76
[12].李静蔚,宋爱莉,张敬涛等.DMBA诱导大鼠乳腺癌癌前病变的组织病理学研究.中华中医药学刊,2008,(3):26
[13].Henry J.Thompson,John N.McGinley,Pamela Wolfe,et al.Temporal sequence of mammary intraductal proliferations,ductal carcinomas in situ and adenocarcinomas induced by 1-methy-1-nitrosourea in rats.Carcinogenesis vol,1998,19(12):21 81-2185
[14].Sue C Heffelfinger,Mei Yan,Robin B Gear,et al.Inhibition of VEGFR2 prevents DMBA-induced mammary tumor formation.Laboratory nvestigation,2004,84:989-998
[15].P.M.Gullino,H.M.Pettigrew,F.H.Grantham.N-nitrosomethylurea as mammary gland carcinogen in rats.Natl.Cancer Inst,1975,54:401-414
[16].Colditz GA,Hankinson SE,Hunter DJ,et al.The use of estrogens and progestins and the risk of breast cancer in postmenopausal women.N Engl J Med,1995,332:1589-1593
[17].V K Turan,R I Sanchez,J J Li,et al.The effects of steroidal estrogens in ACI rat mammary carcinogenesis:17β-estradiol,2-hydroxyestradiol,4-hydroxyestr- adiol,16α-hydroxyestradiol,and 4-hydroxyestrone.Journal of Endocrinology,2004,183:91-99
[18].余望贻,姜海斌,孟琼等.乳腺增生病动物模型的实验研究.中国实验动物学报,2004,(4):12
[19].Harvey JM,Clark GM,Osborne CK,et al.Estrogen receptor status by immunohistochemistry is superior to the lig-and-binding assay for predicting response to adjuvant endocrine therapy in breast cance.J Clin Oncol,1999,17(5):1474-1481
[20].Lawson JS,Field AS,Champion S,et al.Low oestrogen receptor alpha expression in normal breast tissue underlieslow breast cancer incidence in Japan.Lancet,1999,354(9192):1787-1788
[21].Allred DC,Mohsin SK,Fuqua SA.Histological and biological evolution of human premalignant breast disease.Endocr Relat Cancer,2001,8(1):47-61
[22].Shoker BS,Jarvis C,Sibson DR,et al.Oestrogen receptorexpression in the normal and pre-cancerous breast.JPathol,1999,188(3):237-244
[23].Skliris GP,Munot K,Bell SM,et al.Reduced expression of oestrogen using DNA methyl transferase inhibitors in a cell line model.J Pathol,2003,201(2):213-220
[24].Shaban AM,Neill PA,Davies MP,et al.Declining estrogen receptorexpression defines malignant progression of human breast neoplasia[J].Am J Surg Pathol,2003,27(12);1502-1512
[25].Anderson E.The role of oestrogen and progesterone recep-tors in human mammary development and tumorigenesis.Breast Cancer Res,2002,4(5):197-201
[26].Quenel N,Wafflart J,Bergonie F.The prognostic value of CerbB-2 in primary breast carcinoma.A study on 942 cases.Breast Cancer Research and Treatment,1995,3:283-285
[27].浦勇顺,周金良,王芳等.p53和C-erbB-2过表达与乳腺癌预后的关系.上海第二医科大学学报,2001,21(2):170
[28].LiBJ,Zhu ZH,Wang JY.Expression correlation of Ki67 to P53,VEGF,and cerbB-2 genes in breast cancer and their clinicalsignifican.Ai Zheng,2004,23(10):1176-1179
[29].Bhatavdekar JM,PatelDD,ShalNG.Prognostic significance of immunohisto- chemically localized biomarkers in stage Ⅱ and stage Ⅲbreast cancer:a multivariate.Ann Surgoncol,2000,7(4):305-311
[30].RangerGS,JewellA,ThomasV,et al.Elevated expression of COX-2 in breast cancer and ductal carcinoma in situ has no correlation with established prognostic markers. J Surg Onco, 2004, 88(2): 100-103
[31]. Nakopoulou L, Mylona E, Papadaki I, et a. Overexpression of cyclooxygenase-2 is associated with a favorable prognostic phenotype in breast carcinoma. Pathobiol- ogy, 2005, 72(5): 241-249
[32]. Leivonen S K, Kahari V M. Transforming growth factor-beta signaling in cancer invasion and metastasis. Int J Cancer, 2007 , 121 (10) : 2119-2124
[33]. Desruisseau S, Palmari J, Giusti C, et al. Determination of TGF-betal protein level in human primary breast cancers and its relationship with survival. Br J Cancer, 2006, 94(2): 239-246
[1].Gullino PM,Pettigrew HM,Grantham FH.N-nitrosomethylureaas mammary gland carcinogen in rats.J Natl Cancer Inst.,1975,54:401-414
[2].Welsch CW.Host factors affecting the growth of carcinogen-induced rat mammary carcinomas:A review and tribute to Charles Brenton Huggins.Cancer Res,1985,45:3415-3443
[3].Harvell DME,Strecker TE,Tochacek M,et al.Rat strainspecic actions of 17b-estradiol in the mammary gland:Correlation between estrogen-induced lobuloalveolar hyperplasia and susceptibility to estrogen-induced mammary cancers.Proc Natl Acad Sci USA,2000,97:2779-2784
[4].Li SA,Liao JD,Li JJ.Estrogen-induced breast cancer in female ACI rats.New York:Springer-Verlag,2000,20:178-188
[5].Colerangle JB,RoyD.Perturbation of cell cycle kinetics in the mammary gland by stilbene estrogen,diethylstilbestrol.Cancer Lett,1995,94:55-63
[6].Xie B,Tsao SW,Wong YC.Induction of high incidence of mammary tumor in female Noble rats with a combination of 17b-oestradiol and testosterone.Carcinogenesis 1999,20:1069-1078
[7].P.M.Gullino,H.M.Pettigrew,F.H.Grantham.N-nitrosomethylureaas mammary gland carcinogen in rats.J.Natl.Cancer Inst,1975,54:401-414
[8].饶金才,李兰珍,陈云生等.乳腺增生病动物模型的复制及病理类型.中国病理生理杂志,1992,(6):8
[9].甘长清,吴凯南,苏新良等.三苯氧胺对大鼠乳腺增生形态学的影响.重庆医科大学学报,2003,(4):28
[10].吴建新,陆德铭,李道坊等.“乳块消”治疗乳腺增生病的临床及实验研究.上海中医学院,上海市中医药研究院学报,1989,(1):17
[11].余望贻,姜海斌,孟琼等.乳腺增生病动物模型的实验研究.中国实验动物学报,2004,(4):12
[12].吴在德.外科学.第5版.北京:人民卫生出版社,2002,355
[13].姚泰.生理学.第5版.北京:人民卫生出版社,2000,12-413
[14].Djuana M.E.Harvell,Tracy K Strecker,Martin Tochacek,et al.Rat strain-specific actions of 17β-estradiol in the mammary gland:Correlation between estrogen-induced lobuloalveolar hyperplasia and susceptibility to estrogen-induced mammary cancers.Medical Sciences,2000,97(6):2779-2784
[15].Irma H.Russo,Jose Russo.Mammary Gland Neoplasia in Long-Term Rodent Studies.Environmental Health Perspectives,1996,104(9):938-967
[16].Welsch CW,Medina D,KidwellW,et al.Rodent models to examine invivo hormonal regulation of mammary gland tumorigenesis.NewYork.Plenum Press,1987,63-179
[17].Callahan R,Neville MC,DanielCW,et al.Retrovirus and proto-oncogene involvement in the etiology of breast neoplasia.New York:Plenum Press,1987,323-351
[18].Teich N,Wyke J,Mak T,et al.Pathogenesis of retrovirus-induced disease.NY:Cold Spring Harbor Laboratory,1982,20:785-998
[19].MacKenzie WF,Garner FM.Comparison of neoplasms in six sources of rats.J Nad CancerInst,1973,(50):1234-1257
[20].Haseman JF,HuffJ,Boorman GA.Use of historicalcontrol data in carcinogenicity studiesin rodents.Toxicol Pathol,1984,(12):126-135
[21].Ross MH,Bras G.Lasting influence of earlycaloric restriction on prevalence of neoplasms in the rat.J Natl Cancer Inst,1971,(47):1095-1113
[22].Tarone RE,Chu KC,Ward JM.Variability inthe rates of some common naturally occurringtumors in Fischer 344 rats and(C57BL/6N x C3H/HeN)F1(B6C3F1) mice.J Natl CancerInst,1981,(66):1175-1181
[23].杨光伦,姚棒祥.外源性雌激素及三苯氧胺对大鼠乳腺癌癌前期病变的作用.中华普通外科杂志,2001,(2):16
[24].Russo J,Gusterson BA,Rogers AE,et al.Comparativestudy of human and rat mammary tumorigenesis.Lab Invest,1990,(62):1-32
[25].Henry J.Thompsonl,John McGinley,Kim Rothhammer,et al.Ovarian hormone dependence of pre-malignant and malignant mammary gland lesions induced in pre-pubertal rats by 1-methyl-1-nitrosourea.Carcinogenesis,1998,19(3):383-386.
[26].Russo J,Gusterson BA,RogersAE,et al.Comparative study of human and rat mammary tumorigenesis.Lab Invest,1990,62(3):244-278
[27].S A Li,S J Weroha,O Tawfikl,et al.Prevention of solely estrogen-induced mammary tumors in female ACI rats by tamoxifen:evidence for estrogen receptor mediation.J Endocrinol,2002,175(2):297-305
[28].Sue C Heffelfinger,Mei Yan,Robin B Gear,et al.Inhibition of VEGFR2prevents DMBA-induced mammary tumor formation.Laboratory Investigation,2004,(84):989-998
[29].李静蔚,宋爱莉,张维东等.乳腺非典型增生病与血管生成关系的实验研究.肿瘤防治杂志,2005,(12):13
[30].Dawson PJ,Wolman SR,Tait L,et al.MCFIOAT:a model for the evolution of cancer from proliferative breast disease.Am J Pathol,1996,148(1):313-319
[31].Miller FR.Xenograft Models of Premalignant Breast Disease.J Mammary Gland Biol Neoplasia,2000,5(4):379-391
[32].HeppnerGH,Wolman SR.MCF—lOAT:A model for human breast cancer development.Breast J,1999,5(2):122-129
[33].Shull JD,Spady TJ,Snyder MC,et al.Ovary-intact,but not ovariectomized female ACI rats treated with 17-estradiol rapidly develop mammary carcinoma.Carcinogenesis,1997,18:1595-1601
[34].傅西林,李树玲,范宇等.乳腺癌癌前病变与乳腺癌相关的多指标病理学研究.中国肿瘤临床,1999,(1):26
[35].Henry J.Thompsonl,3 and meenakshi sing h2 rat models of premalignant breast disease.J.Cancer,2006,20:985-994