【作者】 刘芳芳；

【导师】 付丽； 郎荣刚； 范宇； 谷峰；

【作者基本信息】 天津医科大学 ， 病理学与病理生理学， 2008， 硕士

【摘要】 【目的】具有高度淋巴结转移能力和不良预后的乳腺浸润性微小乳头状癌(Invasivemicropapillary carcinoma,IMPC)是研究高转移乳腺癌的良好模型,本研究旨在明确趋化因子SDF(stromal cell-derived factor)-1及其受体CXCR4在这种高淋巴结转移的特殊类型乳腺癌中的表达方式、强度及定位,及其与淋巴结转移的关系:通过对比SDF-1/CXCR4在乳腺浸润性微小乳头状癌和髓样癌(medullarycarcinoma,MC)中的表达及其与肿瘤微环境中淋巴细胞浸润的关系,初步探讨不同类型乳腺癌微环境中淋巴细胞浸润的原因及意义;并分析SDF-1/CXCR4在浸润性乳腺癌中的表达与其相关临床病理指标及淋巴结转移之间的关系,验证在特殊类型乳腺癌IMPC中得出的相关结论在其它类型浸润性乳腺癌中的普遍意义。【方法】采用免疫组织化学方法检测103例IMPC(包括IMPC相应淋巴结、对照组96例浸润性导管癌)、34例典型MC及120例浸润性乳腺癌(其中包括上述96例浸润性导管癌及其他类型浸润性乳腺癌)中SDF-1/CXCR4蛋白的表达情况;采用地高辛标记的寡核苷酸探针进行原位杂交以检测SDF-1和CXCR4 mRNA的表达;观察各类乳腺癌组织间质淋巴细胞浸润、淋巴结转移情况等病理学特征,分析SDF-1/CXCR4的表达与各病理学特征之间的关系。【结果】1.IMPC成分在肿瘤中所占比例与淋巴结转移程度无显著相关性(P=0.407);73例淋巴结阳性的混合型IMPC中,与淋巴结转移灶中的其它肿瘤成分相比,IMPC成分是主要的转移成分(P＜0.001)。2.免疫组化及原位杂交显示SDF-1的蛋白及mRNA均主要定位于肿瘤细胞(96%),肿瘤间质细胞也有较弱的表达(14%),且上述两部位的表达均与IMPC淋巴结转移正相关(P＜0.05)。3.罕见SDF-1 mRNA表达的脉管内皮却可见SDF-1蛋白的表达,其表达程度与肿瘤细胞SDF-1胞质着色正相关(P=0.001)。其中淋巴管内皮SDF-1的着色与淋巴结转移程度正相关(P=0.001);血管内皮SDF-1的着色与肿瘤环境中的淋巴细胞浸润正相关(P=0.024),且同时伴有较多淋巴细胞浸润(+/++)及SDF-1血管内皮着色(+)的病例,其淋巴结的转移程度分别高于仅有上述条件之一或二者均不具备的各组病例(P＜0.05)。4.上述免疫组化显示SDF-1阳性的各着色部位中,肿瘤细胞胞质和脉管内皮SDF-1的表达高于IDC组(P=0.001;P=0.019)。5.51例伴有DCIS的IMPC病例中DCIS成分的SDF-1胞膜着色程度明显高于IMPC成分(P=0.009);而胞质SDF-1的着色在两种成分中的表达差异无统计学意义(P＞0.05)。6.CXCR4在IMPC中主要表达于肿瘤细胞的胞核及胞质,仅有胞质的表达与淋巴结受累程度正相关(P=0.008),并且高于IDC组胞质CXCR4的表达(P=0.049)。7.淋巴结转移成分中包含IMPC成分的84例IMPC中,CXCR4在IMPC原发部位胞核和胞质的表达均高于转移淋巴结(P＜0.001)。8.混合型IMPC中IMPC成分肿瘤细胞SDF-1及CXCR4的表达高于内对照(伴随的其他类型的浸润性癌)肿瘤细胞的表达(P=0.003;P=0.001)。9.伴较多淋巴细胞浸润(+/++)的IMPC(n=54),其淋巴结转移程度、SDF-1在肿瘤细胞胞质及间质脉管内皮的表达、CXCR4在肿瘤细胞核的表达均高于MC组(P＜0.01);SDF-1在肿瘤细胞胞膜的表达、CXCR4在肿瘤细胞胞质的表达与MC组差异无统计学意义(P＞0.05)。10.SDF-1在浸润性乳腺癌中主要表达于肿瘤细胞(包括胞质与胞膜);肿瘤细胞胞质SDF-1的表达在淋巴结阳性组高于阴性组(P＜0.05),且其表达程度与淋巴结受累数目、病理学分期、组织学分级、肿瘤大小及ER表达等指标呈正相关(P＜0.05)。11.CXCR4在浸润性乳腺癌中也主要表达于肿瘤细胞(包括胞质与胞核);肿瘤细胞胞质CXCR4的表达在淋巴结阳性组高于阴性组(P＜0.01),且其表达程度与淋巴结受累数目、病理学分期、组织学分级、肿瘤大小及HER-2表达等呈正相关(P＜0.05),而胞核的表达仅与PR的表达情况呈正相关(P＜0.001)。12.肿瘤细胞胞质CXCR4与SDF-1的表达呈正相关(P＜0.01)。【结论】1.IMPC的生物学行为并不取决于肿瘤中IMPC成分的多少。2.SDF-1/CXCR4可能是IMPC结构的分子特征之一,并可能为IMPC高淋巴结转移的原因之一。3.SDF-1和CXCR4在肿瘤组织中的多种定位和不同的表达方式中,SDF-1和CXCR4在肿瘤细胞胞质的表达、SDF-1在脉管内皮的着色可能是引起IMPC高度淋巴结转移的关键因素。4.大量淋巴细胞浸润在乳腺IMPC和MC两种癌中的意义不同。5.IMPC肿瘤微环境中与SDF-1表达相关的淋巴细胞浸润可能促进了其淋巴结转移。6.均伴较多淋巴细胞浸润的IMPC与MC中SDF-1的表达差异可能是二者预后差异的关键因素。7.SDF-1/CXCR4在肿瘤细胞胞质的表达与浸润性乳腺癌的淋巴结转移等多项临床病理指标之间密切相关,可能作为预测乳腺癌淋巴结转移及预后的免疫病理学指标。更多还原

【Abstract】 Objective:Invasive micropapillary carcinoma(IMPC)of the breast is a newly recognized subtype of breast cancer characterized by a high propensity for lymph node metastasis.The purpose of this study is to investigate the relationship between the features of location,expression pattern and intensity of SDF-1/CXCR4 and lymph node metastasis in IMPC of the breast;To explore the reason and the significance of the lymphocytes infiltration in different tumor microenvironment by comparing SDF-1 expression and its relationship with lymphocytes infiltration in IMPC with that in MC of the breast.To investigate the reason of some IMPC in which dense lymphocyte infiltrating but with poor prognosis;To explore the expression of SDF-1/CXCR4 and their association with clinic-pathological features and lymph node metastasis in human invasive breast carcinoma.Methods:Immunohistochemistry and in situ hybridization were performed to analyze the expression of SDF-1/CXCR4 in IMPC(including primary tumor and lymph nodes of IMPC,and 96 cases of IDC as control group),34 of MC and 120 cases of invasive breast cancer.Pathological features,lymphocytes infiltration and status of lymph node metastasis were examined by microscopic analysis.Results:1.The percentage of IMPC component in the whole tumor was not significantly associated with the extent of lymph node metastasis(P=0.067).In 73 cases of mixed-IMPC,IMPC component are the main source for lymph node metastasis compared with other components in the primary tumor(P＜0.001).2.SDF-1 was mainly expressed in tumor cells(96%)and also in stromal cells (14%),staining scores of which were both correlated with the extent of lymph node metastasis in IMPC(P＜0.05).3.Although SDF-1 mRNA was rarely detected in the endothelial cells,SDF-1 immunostaining on lymphatic vessels(36%of IMPCs)was also associated with nodal involvement(P=0.001)and that on blood vessels(34%)correlated with stromal lymphocyte infiltration in IMPC(P=0.024).4.Cytoplasmic SDF-1 expression in tumor cells and SDF-1 staining on vessels in IMPCs were higher than IDCs(P=0.001;P=0.019).5.51 of the 103 IMPCs contained DCIS as or as a part of nonmicropapillary components,in which significantly higher membranous(P=0.009)but similar cytoplasmic SDF-1 staining(P＞0.05)was found compared with IMPC component.6.CXCR4 was detected in either cytoplasm or nucleus of tumor cells,but only cytoplasmic expression in IMPC was correlated with nodal involvement (P=0.008)and higher than IDC group(P=0.049).7.The degree of cytoplasmic and nuclear CXCR4 expression in primary tumor was significantly higher than that in metastatic foci of the same case(P＜0.001).8.SDF-1 and CXCR4 expression in IMPC component were higher than the internal control in mixed-IMPC(P=0.003;P=0.001).9.In the cases of IMPC with abundant infiltrating lymphocytes(+/++)(n=54),the degree of lymph node metastasis,SDF-1 staining in tumor cell cytoplasma and on vessels,CXCR4 nuclear staining were higer than that in MC group(P＜0.01); No difference of SDF-1 membrane staining and CXCR4 cytoplasmic staining was found between the two.groups(P＞0.05).10.SDF-1 was mainly expressed in tumor cells(both membrane and cytoplasm); Cytoplasmic SDF-1 expression in lymph node-positive group was higher than that in node-negative group,and the level of which was correlated with the number of nodal involvement,TNM stages,histological grades,tumor diameter and ER status(P＜0.05).11.CXCR4 was also mainly expressed in tumor cells(both nuclei and cytoplasm); Cytoplasmic SDF-1 expression in lymph node-positive group was higher than that in node-negative group,and the level of which was correlated with the number of nodal involvement,TNM stages,histological grade,tumor diameter and HER-2 status(P＜0.05),but the nuclear expression was only correlated with PR status(P＜0.001).12.The cytoplasmic CXCR4 was correlated with SDF-1 expression positively (P＜0.01).Conclusion:1.The biological behavior of IMPC is not dependent on the percentage of IMPC components in the whole tumor.2.High expression of SDF-1/CXCR4 might be one of the molecular characteristics of IMPC and facilitate lymphatic metastasis.3.SDF-1 located in tumor cells and on vessels,cytoplasmic staining pattern of SDF-1 and CXCR4 may be the key factors that influence the extent of lymph node metastasis in IMPC.4.The significance of lymphocyte infiltrating for prognosis might be different.5.Abundant infiltrating lymphocytes which associated with SDF-1 expression in tumor microenvironment of IMPC might he a factor promoting lymph node metastasis.6.The difference of SDF-1 expression in the two types of breast carcinoma might be the key factor leading to their different prognosis.7.The potential value of SDF-1 and CXCR4 as biomarkers for diagnosis and for predicting lymph node metastasis and a potential therapeutic target in invasive breast carcinoma,but different location in tumor microenvironment and staining patterns may signify different features of tumors.更多还原