【作者】 王芳；

【导师】 付占昭；

【作者基本信息】 河北医科大学 ， 肿瘤学， 2009， 硕士

【摘要】 背景和目的:乳腺癌是妇女最常见的恶性肿瘤之一,近年来在我国,特别是大城市内的发病率仍呈直线上升趋势。乳腺癌已被视为全身性疾病,化疗是治疗乳腺癌的重要手段之一,但化疗耐药的出现,导致患者无病生存期缩短,所以探讨有关乳腺癌耐药相关基因,指导规范化治疗尤为重要。肿瘤细胞微环境的改变与肿瘤多药耐药的关系成为目前研究的新热点。肿瘤细胞微环境改变中最重要的是细胞的缺血缺氧,由于缺氧诱导因子-l(hypoxia-inducible factor-1, HIF-1)在细胞缺氧改变中占有重要中心地位,因此作为其活性亚单位的HIF-1α与肿瘤细胞多药耐药的关系备受关注。P-糖蛋白(permeability- glucoprotein, P-gp)过表达是肿瘤多药耐药(multidrug resistance, MDR)产生的一个主要原因,拓扑异构酶Ⅱα(topoisomeraseⅡα, Topo-Ⅱα)是多种抗肿瘤药物如蒽环类药物的靶分子,其表达降低将影响肿瘤对化疗的敏感性。本研究采用免疫组织化学方法检测HIF-1α、P-gp与Topo-Ⅱα在乳腺癌组织中的表达,分析其表达与患者临床病理特征及三者之间的相关关系,探讨缺氧对乳腺癌发生、发展以及化疗耐药的影响,希望对乳腺癌患者的治疗及预后提供可靠的参考指标,以及有助于实现逆转肿瘤耐药和提高化疗效果。方法:选取秦皇岛市第一医院2000年9月~2003年12月接受手术治疗的乳腺癌患者的组织石蜡标本59例,采用免疫组织化学SP法对乳腺癌组织HIF-1α、P-gp与Topo-Ⅱα的表达进行检测,并结合完整的临床资料,采用SPSS14.0软件系统进行分析,应用χ2检验等分析三者与临床病理特征的关系,以p<0.05为有统计学意义。结果:1 HIF-1α、P-gp与Topo-Ⅱα在59例乳腺癌组织中的阳性表达率分别为:61.01%(36/59)、40.68%(24/59)和49.15%(29/59),而在15例癌旁正常乳腺组织中均为阴性表达,HIF-1α、P-gp与Topo-Ⅱα在乳腺癌组织中的表达均明显高于正常乳腺组织,差异具有统计学意义(p<0.05)。2 HIF-1α、P-gp与Topo-Ⅱα的表达与乳腺癌不同的临床病理特征之间的关系:HIF-1α在T≤2cm、2cm<T≤5cm和T>5cm组中的阳性表达率分别为30.77%(4/13)、65.71%(23/35)和81.82%(9/11),各组差异均具有统计学意义(P<0.05);随着乳腺癌组织T分期的增加HIF-1α的阳性表达率增高,具有线性趋势(P<0.05)。HIF-1α在组织学Ⅰ、Ⅱ和Ⅲ级组中的阳性表达率分别为22.22%(2/9)、58.62%(17/29)和80.95%(17/21),各组间差异均具有统计学意义(P<0.05);随着乳腺癌组织学分级的增加HIF-1α的阳性表达率增高,具有线性趋势(P<0.05)。HIF-1α在有淋巴结转移组中的阳性表达率明显高于其在无淋巴结转移组中的阳性表达率(76.47%VS40.00%),差异具有统计学意义(P<0.05)。HIF-1α在病理分期0~Ⅰ、Ⅱ和Ⅲ期组中的阳性表达率分别为58.33%(7/12)、61.11%(22/36)和63.64%(7/11),三组间差异无统计学意义(P >0.05);在不同年龄、病理类型及月经状态组中HIF-1α阳性表达率的差异无统计学意义(P >0.05)。P-gp在有淋巴结转移组中的阳性表达率明显高于其在无淋巴结转移组中的阳性表达率(52.94%VS24.00%),差异具有统计学意义(P<0.05);在不同年龄、肿块大小、组织学分级、病理分期、病理类型及月经状态组中P-gp阳性表达率的差异均无统计学意义(P >0.05)。在不同年龄、肿块大小、组织学分级、淋巴结转移情况、病理分期、病理类型及月经状态组中Topo-Ⅱα阳性表达率的差异均无统计学意义(P >0.05)。3乳腺癌中HIF-1α与P-gp表达的相关性HIF-1α阳性组的P-gp阳性表达率为55.56%(20/36),HIF-1α阴性组的P-gp阴性表达率为82.61%(19/23),HIF-1α与P-gp表达呈正相关表达(r=0.38,P<0.05)。4乳腺癌中HIF-1α与Topo-Ⅱα表达的相关性HIF-1α阳性组的Topo-Ⅱα阳性表达率为55.56% (20/36),HIF-1α阴性组的P-gp阴性表达率为60.87%(14/23),HIF-1α与Topo-Ⅱα表达无相关性(r=0.16,P >0.05)。结论:1 HIF-1α表达与乳腺癌肿块大小、组织学分级和淋巴结转移与否呈正相关,HIF-1α过表达提示乳腺癌具有更高的侵袭性。2 P-gp的表达与乳腺癌淋巴结转移与否呈正相关,可能是乳腺癌复发、转移的一个高危因素。3 Topo-Ⅱα在乳腺癌组织中过表达,而在正常乳腺组织中呈阴性表达,且与临床病理特征之间无相关性,提示它可能是一个独立的影响耐药的因素。4 HIF-1α与P-gp在乳腺癌中的表达呈正相关,提示其可能与乳腺癌多药耐药相关,可以为揭示耐药机制提供新的思路。5 HIF-1α与Topo-Ⅱα在乳腺癌中的表达无相关性。更多还原

【Abstract】 Background and Objective: Breast cancer is the most common malignant disease of women. Recently, the mortality of breast cancer is sharp rising in China, especially in big citys. Breast cancer has already been regarded as a systemic disease, and chemotherapy is one of the important therapeutic approaches to treat breast cancer. But the disease free survival (DFS) of patients has been induced by the appearance of drug resistant, so it is important to probe the breast cancer resistant related gene and guide the standard therapeutics. The relationship between the changes of cell microenvironment and multidrug resistance (MDR) becomes a new hot spot. The most important change of cell microenvironment is the ischemia and hypoxia. Since hypoxia-inducible factor-1 (HIF-1) plays an important role in cell microenvironment changes,it is very interesting to investigate the relationship between MDR and HIF-1α—the active subunit of HIF-1. The over expression of permeability-glycoprotein (P-gp) plays a more important role in MDR, The DNA topoisomerase-Ⅱ(Topo-Ⅱ) is the effective target of many anti-tumor medicine including anthracycline, and it is often associated with drug resistance when its level decrease. This study was designed to evaluate the expression of HIF-1αand P-gp, Topo-Ⅱαin breast cancer by immunohistochemistry and investigate the relationships between their expressions and the clinicopathological characteristics, and we also analysed the association among their expressions,which may reveal the role of hypoxia in the pathogenesis and progression of breast cancer. Thus we would find a promising target for therapy and a potentially useful factor for prognostic detection in breast cancer,which would retroconverse tumor drug resistance and improve the effect of chemotherapy.Methods: We collected 59 breast cancer samples which were resected by surgery and confirmed by pathology in the First Hospital of Qin Huangdao from September, 2000 to December, 2003. We detected the expression of HIF-1α, P-gp and Topo-Ⅱαusing immunohistochemieal SP methods to investigate the relationship between them,as well as the correlate with clinicopathological characteristics in human breast cancer. Statistical methodsχ2 test was performed by SPSS software version 14.0.Results: 1 Expression of HIF-1α, P-gp and Topo-Ⅱαin breast cancer: The positive rates of the expression of HIF-1α, P-gp and Topo-Ⅱαin 59 cases were 61.01%、40.68% and 49.15% respectively. These proteins were all negative in 15 normal breast tissues. And there were significant differences between breast cancer tissues and normal tissues (P<0.05).2 The relationship of the expression of HIF-1α, P-gp and Topo-Ⅱαand the clinicopathological characteristics:Expression of HIF-1αwas positively related to tumor size, histological gradesand and lymph node matastasis, and had linear trend with tumor size and histological grades (P<0.05). And it was not correlated with patients’age, pTNM stages, histological types and menstruation status (P>0.05).The positive rate of the expression of P-gp in lymph node matastasis group (52.94%) was higher than that in no lymph node matastasis group (24.00%), the difference had statistical significance (P<0.05). The expression of P-gp had significantly positive correlation with lymph node matastasis (P<0.05), and no correlation with age, tumor size, histological grades, pTNM stages, histological types and menstruation status (P>0.05).There was no correlation between the expression of Topo-Ⅱαand any of the clinicopathological characteristics mentioned as above, such as age,tumor size, histological grades, et al (P>0.05).3 The relationship between expression of HIF-1αand P-gp: The positive expression rate of P-gp was 55.56%(20/36) in the group of HIF-1αpositive. The negative expression rate of P-gp was 82.61%(19/23) in the group of HIF-1αnegative. Expression of P-gp was positively correlated with HIF-1α(r=0.38, P<0.05). 4 The relationship between expression of HIF-1αand Topo-Ⅱα: There was no correlation between the expression of Topo-Ⅱαand HIF-1α(r=0.16, P >0.05).Conclusions: 1 The expression of HIF-1αis positively related to tumor size, histological gradesand and lymph node matastasis, its over expression shows that the invasion of breast cancer is even higher.2 The expression of P-gp has significantly positive correlation with lymph node matastasis, it may be a high risk factor for recurrence and matastasis of breast cancer.3 Topo-Ⅱαis over expressed in breast cancer tissues, and is negative in normal tissues, it may be a particular factor of drug resistance.4 The expression of HIF-1αis positively correlated with P-gp, and HIF-1αmay play an important role in multidrug resistance, which may provide a novel application strategy gene targeted therapy of breast cancer.5 There is no correlation between the expression of HIF-1αand Topo-Ⅱαin breast cancer.更多还原