乳腺癌中Caveolin-1蛋白的表达及临床意义

目的 观察窖蛋白(Caveolin-1,Cav-1)在正常乳腺组织、良性增生性乳腺病及乳腺癌中的表达及其与乳腺癌临床病理特征的关系.方法 采用免疫组化SP法检测10例正常乳腺组织、15例良性增生性乳腺病和48例乳腺癌组织中Cav-1的表达,分析Cav-1在不同乳腺病变中表达的差异.结果 Cav-1在正常乳腺组织、良性增生性乳腺病及乳腺癌中的阳性率分别为40%、46%和100%,平均表达强度积分(±s)分别为0.90±0.83、0.93±1.16和6.24±2.18.其在正常乳腺组织与良性增生性乳腺病之间表达差异无统计学意义(P>0.05);在正常乳腺组织、良性增生性乳腺病与乳腺癌之间表达差异具有显著性(P<0.01);在乳腺癌不同组织学类型、病理分级、临床分期、细胞增殖指数(MIB-1/Ki-67)和有无淋巴结转移中阳性率和表达强度差异均无显著性(P>0.05).结论 Cav-1在正常乳腺组织及良性增生性乳腺病中呈低表达,在乳腺癌中呈高表达,表明Cav-1与乳腺癌的发生密切相关,但与乳腺癌的组织学类型、病理分级、临床分期、有无淋巴结转移和细胞增殖指数(MIB-1/Ki-67)无明显相关性.     

杂交瘤细胞免疫电泳法检测乳腺癌患者血清BG6抗原的初步研究

用自建的杂交瘤细胞免疫电泳测定法,对乳腺癌患者血清BG6抗原的表达特性进行了初步研究。检测对象170例。包括正常人50例,乳腺癌42例,其他癌57例,乳腺良性病21例。实验用杂交瘤为自制的分泌抗人乳腺癌单克隆抗体的杂交瘤细胞株BG6,为便于保存,先用0.025％     

神经细胞黏附分子在老年乳腺癌中表达的研究

目的:研究神经细胞黏附分子(neural cell adhesion molecules,NCAM)在乳腺肿瘤中的表达并讨论其临床意义。方法:选择老年(60~72岁)乳腺癌组织25例(神经侵袭的8例,无神经侵袭的17例)、癌旁组织25例,乳腺良性病变26例(乳腺纤维腺瘤16例,乳腺腺病4例,囊性乳腺病6例),用免疫组织化学方法检测NCAM在这些组织中的表达。用酶联免疫吸附法(ELISA)测定NCAM在乳腺癌、乳腺良性病病和正常对照(n=30,61~68岁)血清中含量。结果经统计学处理。结果:NCAM在部分乳腺癌细胞质和细胞膜呈宗黄色阳性表达,癌旁组织和良性病变表达很少。乳腺癌组、癌旁组及乳腺良性病变NCAM阳性率分别为28.0%(7/25),4.0%(1/25)和3.8%(1/26),腺癌组织明显高于癌旁组和乳腺良性病变(P0.05)。NCAM阳性表达在乳腺癌有神经侵袭病例阳性率50.0%(4/8)明显高于无神经侵袭病例23.5%(4/17,P0.05)。血清NCAM浓度(pg/ml)结果,乳腺癌(69.8±29.4)显著高于乳腺良性病变(16.7±6.3)和正常对照(14.9±3.1),具有明显统计学差异(P0.05)。结论:NCAM在老年乳腺癌有明显表达,且伴发神经侵袭和转移时表达更为明显,表明NCAM表达影响乳腺癌发生发展,这为临床防治老年乳腺癌提供重要的参考依据。     

活检乳腺肿物与McAb SC3A反应的免疫组化研究

本文用自制的McAbSC3A对14种153例不同组织类型的乳腺癌活检组织对应抗原的表达进行了免疫组化的研究。 活检组织以10％福尔马林固定。石蜡包埋,切片5μm厚,HE染色后经病理确诊为乳癌80例,乳腺纤维瘤32例、乳腺增生性病变(乳腺病)29例、乳腺导管内乳头状瘤2例、浆细胞性乳腺炎2例和男子乳腺发育8例。免疫组化染色以PAP法进行,并以 3等4级法记分。 结果表明,SC3A与乳癌结合的阳性率     

AKT-1在乳腺良恶性病变中的表达及其临床意义

目的通过研究AKT-1在乳腺癌、癌旁正常组织、乳腺纤维腺瘤中的表达情况,探讨其在乳腺癌评估预后及治疗方面的价值。方法采用免疫组化SP法,检测61例乳腺癌、20例癌旁正常组织、20例乳腺纤维腺瘤中AKT-1的表达情况。结果 AKT-1在乳腺癌旁正常组织中无表达,在纤维腺瘤中低表达(阳性率为15%),在乳腺癌组织中高表达(阳性率为49.2%),差异具有统计学意义(P<0.05)。在乳腺癌组中,AKT-1的阳性表达率与HER-2表达情况呈正相关;在原发灶大小不同、腋窝淋巴结有无转移和TNM分期不同组中,AKT-1的阳性表达率差异有统计学意义(P<0.05);在不同年龄、不同雌孕激素受体情况和不同组织学类型中,差异无统计学意义(P>0.05)。结论 AKT-1可能是导致乳腺癌发生、发展的因素之一,在乳腺癌细胞向周围组织浸润,远处转移中起一定作用,对判断乳腺癌的恶性程度及预后有重要意义。     

TTC25在乳腺癌中的表达及意义探讨

目的探讨TTC25在乳腺癌组织中的表达与乳腺癌临床病理参数的关系。方法应用免疫组化及Western blot方法检测TTC25在正常乳腺组织及浸润性癌组织中的表达与临床病理因素的关系。结果TTC25在浸润性乳腺癌中的表达明显高于癌旁正常乳腺上皮细胞(阳性率为78.75%Vs 33.75%,P0.05),淋巴结转移组的表达率高于非淋巴结转移组的表达率(阳性率为40.74%Vs 69.81%,P0.05),Western blot的方法证明在浸润性乳腺癌中蛋白水平表达量明显高于癌旁正常乳腺组织(P0.05)。结论TTC25可能在乳腺癌的发生发展过程中起一定的促进作用,为目前乳腺癌的靶向治疗提供了一个新的研究靶点。     

乳腺癌中分泌成分的表达及其临床意义

目的探讨分泌成分(secretory component,SC)在良性乳腺腺泡及乳腺癌组织中的表达及两者的相关性。方法采用免疫组化SP法检测SC在114例良性乳腺腺泡中的表达,并检测SC、ER、PR在133例乳腺癌中的表达。结果SC在良性乳腺腺泡和乳腺癌中的阳性率分别为69.3%(79/114)、22.6%(30/133)。SC阳性率与非特殊型浸润性乳腺癌的分化程度无显著相关性(P=0.543);但与浸润性小叶癌的类型相关,其在腺泡型浸润性小叶癌中的阳性率高于多形性、经典型、实性型中的阳性率(P=0.014)。此外,SC阳性率与浸润性小叶癌PR表达状态相关,其在PR阳性患者中的阳性率低于PR阴性患者中的阳性率(P=0.024)。结论SC在乳腺癌中表达明显低于良性乳腺腺泡,且在腺泡型浸润性小叶癌中的表达明显高于其他亚型浸润性小叶癌。SC在浸润性小叶癌中PR阳性患者中的阳性率低于PR阴性患者中的阳性率。     

抗c-erbB-2p185蛋白胞内区多肽单克隆抗体的制备、鉴定及初步应用

目的 研制抗c-erbB-2胞内区多肽单克隆抗体,用于乳腺癌等c-erbB-2过量表达的诊断及指导治疗。方法 合成p185蛋白胞内区多肽作为抗原,免疫BALB／C小鼠建立了一组分泌抗该多肽的特异性高亲和力单克隆抗体杂交瘤细胞系,并对该单抗的生化特性及其免疫学反应性进行了研究和测定,并与美国食品药品管理局（FDA）批准的抗c-erbB-2多克隆抗体进行了比较。结果 共获得3株稳定分泌抗p185胞内区单抗的杂交瘤细胞株。其中035－E61株单抗经过39例（T1－2N0M0）乳腺癌患者组织病理切片进行免疫组织化学染色,c-erbB2-2过量表达阳性率为26％,而乳腺纤维腺瘤组织切片没有非特异染色;流产正常胎儿器官组织切片除甲状腺、肾上腺和肾小管上皮表达外,其余均不表达。与FDA批准的DAKO公司多抗的阳性染色符合率为74％。结论 035－E61单抗可特异识别乳腺癌细胞内p185胞内区抗原。对判断乳腺癌预后及指导治疗可能具有应用价值。     

乳腺癌和癌旁乳腺组织中Notch1基因mRNA及蛋白的表达

目的 检测Notch1基因mRNA及Notch1蛋白在人乳腺癌和癌旁乳腺组织中的表达,分析其临床病理学意义.方法 应用逆转录聚合酶链反应(RT-PCR)方法榆测60例乳腺浸润性导管癌和60例癌旁乳腺组织中Notch1基因mRNA,应用免疫组织化学SP法检测60例乳腺浸润性导管癌、30例导管原位癌及60例癌旁乳腺组织Notch1蛋白的表达,分析Notch1表达水平与乳腺癌临床病理特征的相关性.结果 Notch1基因mRNA在人乳腺浸润性导管癌及癌旁乳腺组织中均有表达.Notch1蛋白在癌旁乳腺组织和导管原位癌中的阳性牢分别为55%(33/60)、70%(21/30),二者差异无统计学意义(P>0.05),在乳腺浸润性导管癌中的阳性率为90%(54/60),明显高于癌旁乳腺组织和导管原位癌的阳性率(P<0.05).乳腺浸润性导管癌Notch1蛋白的高表达与肿瘤的淋巴结转移(P=0.006)、病理学分级(P=0.001)和TNM分期(P=0.022)均呈显著正相关.结论 乳腺浸润性导管癌存在Notch1蛋白的高表达.Notch1蛋白高表达与乳腺癌的恶变演进有关.Notch1基因的表达可能影响乳腺癌的发生、发展.     

乳腺癌中PHF2的表达及其临床意义

目的探讨PHF2在乳腺癌细胞和组织中的表达及其与乳腺癌临床病理特征、预后的关系。方法采用q RT-PCR法检测乳腺癌及正常乳腺上皮细胞株(n=7)及配对的癌旁组织(n=30)中PHF2 m RNA的表达水平;应用Western blot法检测乳腺癌细胞株中PHF2蛋白的表达水平,免疫组化法检测乳腺癌组织(n=50)和癌旁组织(n=15)中PHF2蛋白的表达,分析PHF2表达与乳腺癌临床病理特征的关系及预后的关系。结果与正常人乳腺上皮细胞相比,PHF2 m RNA和蛋白在乳腺癌细胞株中表达下调(P0.01);与癌旁组织相比,PHF2 m RNA在90%(27/30)的乳腺癌组织中表达下调(P0.05);免疫组化结果显示:在70%(35/50)的乳腺癌组织中PHF2蛋白表达下调(P0.05);PHF2表达下调与乳腺癌患者的淋巴结转移、ER阴性及预后密切相关(P0.05)。结论 PHF2在乳腺癌中表达下调,可以作为乳腺癌预后判断的指标之一,有望成为乳腺癌治疗的新靶点。     

Prostate-specific membrane antigen (PSMA) expression in breast cancer and its metastases

The present study was undertaken to investigate the expression of prostate-specific membrane antigen (PSMA) in normal breast tissues, in cancerous breast tissues and in distant metastases from patients with breast cancer. Immunohistochemical analysis was performed to determine PSMA expression and angiogenic activity using anti-PSMA mAb and anti-CD31 mAb respectively. Immunofluorescence staining was applied to confirm the exact co-localization of PSMA and CD31. We observed different patterns of PSMA expression between normal and cancerous tissues. Normal breast tissues showed PSMA expression only in normal glandular cells. However, primary breast tumors and distant metastases showed PSMA expression in tumor cells and in tumor-associated neovasculature. PSMA score group status in primary breast tumors was significantly associated with histologic type and tumor grade (p?=?0.026 and p?=?0.004 respectively). Distant metastases showed higher PSMA expression in tumor-associated neovasculature comparing with primary tumors. Moreover, brain tumor-associated neovasculture had significantly higher expression of PSMA comparing with bone tumor-associated neovasculture. The localized binding of PSMA mAb to the neovasculature endothelium was confirmed with the double Immunofluorescence staining. ⁶⁸Ga-PSAM imaging of a patient with metastatic breast cancer showed strong tracer uptake in all known skeletal metastases. To the best of our knowledge, this study is the second one that has assessed PSMA expression in a large number of breast cancer patients. Our findings showed that PSMA is particularly expressed in tumor-associated neovasculature of breast tumors and its distant metastases, thus enhancing the evidence on the potential usefulness of PSMA as a therapeutic vascular target.     

Differential expression of the embryo/cancer gene ECSA(DPPA2), the cancer/testis gene BORIS and the pluripotency structural gene OCT4, in human preimplantation development

In this paper, we examine the expression profiles of two new putative pluripotent stem cell genes, the embryo/cancer sequence A gene (ECSA) and the cancer/testis gene Brother Of the Regulator of Imprinted Sites (BORIS), in human oocytes, preimplantation embryos, primordial germ cells (PGCs) and embryo stem (ES) cells. Their expression profiles are compared with that of the well-known pluripotency gene, OCT4, using a primer design that avoids amplification of the multiple OCT4 pseudogenes. As expected, OCT4 is high in human oocytes, down-regulated in early cleavage stages and then expressed de novo in human blastocysts and PGCs. BORIS and ECSA show distinct profiles of expression in that BORIS is predominantly expressed in the early stages of preimplantation development, in oocytes and 4-cell embryos, whereas ECSA is predominantly expressed in the later stages, blastocysts and PGCs. BORIS is not detected in blastocysts, PGCs or other fetal and adult somatic tissue tested. Thus, BORIS and ECSA may be involved in two different aspects of reprogramming in development, viz., in late gametogenesis, and at the time of formation of the ES cells (inner cell mass (ICM) and PGC), respectively. However, in human ES cells, where a deprogrammed stem cell state is stably established in culture, an immunofluoresence study shows that all three genes are co-expressed at the protein level. Thus, following their derivation from ICM cells, ES cells may undergo further transformation in culture to express a number of embryo and germ line stem cell functions, which, in normal development, show different temporal and spatial specificity of expression.     

Expression and localization of GRP75 in human epithelial tumors and normal tissues.

Using differential display mRNA techniques, the authors found cDNA of the heat shock 70 protein known as GRP75 overexpressed in ovarian cancer cell lines. In the current study, the authors used immunohistochemistry to characterize the expression pattern of GRP75 in ovarian carcinomas and compared it with epithelial tumors originating from the female reproductive tract, epithelial neoplasms from non-gynecologic sites (colon, pancreas, breast, and lung), and various normal tissues. The authors also developed an antigen capture ELISA assay to determine if GRP75 can be detected in tumors, ascites, or sera of patients with advanced mullerian adenocarcinomas. All epithelial tumors from the ovary and the female reproductive tract were positive for GRP75 expression with moderate to strong staining intensity; stromal expression of GRP75 was generally weak or absent. Adenocarcinomas from the colon, lung, pancreas, and breast also stained strongly positive for GRP75. The epithelial cells of all normal tissues examined were positive for GRP75, and strong staining was also seen in the corpora lutea, hepatocytes, enteric neural plexus of the esophagus and colon, and placental cytotrophoblast and syncytiotrophoblast, and in subpopulations of pancreatic acinar cells. The ELISA assay detected GRP75 in tumor lysates and ascitic fluid, but not sera, of patients with mullerian adenocarcinomas. The authors conclude that GRP75 is highly expressed in both benign and malignant epithelium, as well as cells of specialized function from a variety of tissues.     

脂肪酸结合蛋白在人乳腺癌组织的表达及意义

目的研究脂肪酸结合蛋白(FABP)mRNA及蛋白质在浸润性乳腺导管癌和乳腺纤维腺瘤的表达及分布,探寻人浸润性乳腺导管癌新的分子标志,为乳腺癌的靶向治疗提供理论依据。方法用半定量逆转录聚合酶链反应、免疫组织化学染色和Western blot等方法检测脂肪细胞型FABP、心型或骨骼肌型FABP、脑型FABP、表皮或牛皮癣型FABP、肝型FABP、小肠型FABP和胃型FABP mRNA及蛋白质在35例浸润性乳腺导管癌,16例乳腺纤维腺瘤组织中的表达变化。结果RT-PCR结果显示A-、B-、I-和G-FABPs mRNA在两种组织的表达差异无统计学意义(P>0.05);但E-、H-和L-FABP mRNA在浸润性导管癌的表达较纤维腺瘤显著升高(P<0.05)。免疫组织化学染色显示,E-、L-和H-FABP在浸润性导管癌的阳性细胞百分数较纤维腺瘤明显上调(P<0.05),分布范围更广。Western blot分析结果进一步证实,E-、L-和H-FABP蛋白质在浸润性导管癌表达明显上调(P<0.05)。结论E-、L-和H-FABP与浸润性乳腺导管癌的发生发展有关,对进一步探寻浸润性乳腺导管癌的分子标志及治疗途径具有理论意义。     

The meaning of the c-kit proto-oncogene product in malignant transformation in human mammary epithelium

To evaluate the relationship between the c-kit proto-oncogene product and malignant transformation of human breast tissue, we examined the immunohistochemical expression of the c-kit proto-oncogene product in both malignant and non-malignant breast tissues. The immunohistochemical expression of the c-kit proto-oncogene product in 40 primary breast cancer tissues (22 axillary lymph nodes negative, 18 lymph nodes positive), in 18 corresponding axillary lymph nodes, and in 10 distant metastastic tissues were studied using an anti-c-kit proto-oncogene product antibody in comparison with 20 normal and 20 benign breast tissues. The mean values of immunoreactive score (IRS) were compared. The IRS of the c-kit proto- oncogene product in normal mammary epithelia was 5.90±1.37 (mean ± s.d.). In benign tissues, the c-kit proto-oncogene product was detected heterogeneously with a reduced IRS (4.05±1.82). In primary breast cancer tissues, the expression of the c-kit proto-oncogene product was often deleted and the average IRS (0.90±1.73) was significantly reduced compared to those of the normal breast tissues or benign breast disease tissues, but no significant difference was shown between the breast cancer groups. The c-kit proto-oncogene product may correlate with growth control or the differentiation of normal breast epithelium. This result suggests that the loss of expression of this protein might correlate with malignant breast cancer progression, but it is most likely involved at an early stage of human breast cancer development. This revised version was published online in July 2006 with corrections to the Cover Date.     

Immunohistochemical study of D5 antigen (an oestrogen receptor related protein) in normal breast, benign breast disease, and mammary carcinoma in situ.

Patterns of staining with a monoclonal antibody which recognises D5 antigen (a 29,000 kD oestrogen receptor-related protein) were studied in seven normal and 76 benign breast biopsy specimens as well as in 12 cases of pure in situ mammary carcinoma. Staining in benign breast lesions was weak and heterogeneous when compared with that seen in most infiltrating carcinomas. In situ carcinomas showed an intermediate pattern of staining. The finding of only small foci of weak positivity for D5 antigen in normal and benign breast disease indicated that there are similarities between the expression of D5 antigen and the presence of oestrogen receptor protein in these tissues. A further similarity was seen with in situ carcinomas, which have been shown to have lower oestrogen receptor content than infiltrating carcinomas and a more heterogeneous staining pattern with D5 than is seen in infiltrating tumours. The importance of these findings remains to be evaluated because the precise nature of D5 antigen and its association with the oestrogen receptor molecule is not fully understood.     

Quantitative assessment of Tn antigen in breast tissue micro-arrays using CdSe aqueous quantum dots

In this study, we examined the use of CdSe aqueous quantum dots (AQDs) each conjugated to three streptavidin as a fluorescent label to image Tn antigen expression in various breast tissues via a sandwich staining procedure where the primary monoclonal anti-Tn antibody was bound to the Tn antigen on the tissue, a biotin-labeled secondary antibody was bound to the primary anti-Tn antibody, and finally the streptavidin-conjugated AQDs were bound to the biotin on the secondary antibody. We evaluated the AQD staining of Tn antigen on tissue microarrays consisting of 395 cores from 115 cases including three tumor cores and one normal-tissue core from each breast cancer case and three tumor cores from each benign case. The results indicated AQD-Tn staining was positive in more than 90% of the cells in the cancer cores but not the cells in the normal-tissue cores and the benign tumor cores. As a result, AQD-Tn staining exhibited 95% sensitivity and 90% specificity in differentiating breast cancer against normal breast tissues and benign breast conditions. These results were better than the 90% sensitivity and 80% specificity exhibited by the corresponding horse radish peroxidase (HRP) staining using the same antibodies on the same tissues and those of previous studies that used different fluorescent labels to image Tn antigen. In addition to sensitivity and specificity, the current AQD-Tn staining with a definitive threshold was quantitative.     

结肠癌相关抗原的制备及其临床意义

目的从培养的结肠癌细胞LOVO中纯化结肠癌相关抗原,探讨其在结肠癌患者血清中的表达。方法裂解结肠癌细胞LOVO,以抗结肠癌相关抗原单克隆抗体(mAb)4D10作为配基进行亲和层析纯化结肠癌相关抗原,SDS-PAGE电泳和Western blot进行鉴定,并用双抗夹心ELISA法检测该相关抗原在结肠癌患者及正常人血清中的表达。结果纯化所获与4D10特异结合的结肠癌相关抗原,其相对分子质量(Mr)约为65000,该抗原由Mr约为30000和35000两种亚基组成。人血清检测实验表明,该抗原在结肠癌患者血清中有较高的表达,与正常人血清相比存在显著的统计学差异(P<0.01)。结论利用mAb4D10进行免疫亲和层析,从结肠癌细胞LOVO中获得纯化的肿瘤相关抗原,该抗原有可能在临床上为结肠癌的诊断提供一定的参考价值。     

乳癌组织端粒酶活性表达及其与p16基因的关系

目的：探讨乳房恶性肿瘤组织中端粒酶活性的表达情况及ｐ１６抑癌基因的影响。方法：应用端粒重复放大程序－酶标法（ＴＲＡＰ－ＥＬＩＳＡ）及ＴＲＡＰ－银染法检测３５例有乳癌组织及２４例良性肿瘤组织中的端粒酶活性,并采用多重ＰＣＲ技术对乳癌组织的ｐ１６基因的缺失空谈进行了分析,结果：３５例乳癌标本中有２８例端粒酶活性表达为阳性（８０％）,而２４例乳房良性肿瘤标本中仅１例端粒酶活性表达阳性（４．１％）。此外３５例乳癌标本中的１５例存在有ｐ１６基因的外显子２及外显子１的缺失（４２．８％）,而该１５例中有１４例被检测为端粒酶活性阳性（９３．３％）,但２０例未缺失标本的端粒酶阳怀率仅为７０％（１４／２０）。结论：端粒酶活性与乳房肿瘤组织的恶性程度密切相关,提示端粒酶参与了肿瘤的形成过程,而端粒酶活性的增高与ｐ１６抑癌基因的缺失突     

LETM1 overexpression is correlated with the clinical features and survival outcome of breast cancer

Background: Leucine zipper/EF hand-containing transmembrane-1 (LETM1) is a mitochondrial inner membrane protein that was first identified in Wolf-Hirschhorn syndrome. However, high-level expression of LETM1 has been correlated with multiple human malignancies, suggesting roles in carcinogenesis and tumor progression. This study is aimed to explore the clinicopathological characteristics and prognostic value of LETM1 overexpression in breast cancer. Methods: Immunohistochemical (IHC) staining, and immunofluorescence (IF) were performed to examine LETM1 expression in breast cancer cell line/tissues compared with adjacent normal tissues. Statistical analysis was applied to evaluate the correlation between LETM1 overexpression and the clinicopathological features of breast cancer. Survival rates were calculated using the Kaplan-Meier method, and the relationship between prognostic factors and patient survival was analyzed using the Cox proportional hazard models. Results: LETM1 protein showed cytoplasmic staining pattern in breast cancer. The strongly positive rate of LETM1 protein was 61.6% (98/159) in breast cancer, which was significantly higher than in DCIS (29.7%, 11/37), hyperplasia (16.7%, 3/18) and adjacent normal breast tissues (15.9%, 7/44). High-level expression of LETM1 protein was correlated with lymph node metastasis, poor differentiation, late clinical stage, disease-free survival (DFS) and overall survival (OS) rates in breast cancer. Moreover, multivariate analysis suggested that LETM1 emerged as a significant independent prognostic factor along with clinical stage of patients with breast cancer. Conclusions: LETM1 plays an important role in the progression of breast cancer. High level expression of LETM1 is an independent poor prognostic factor of breast cancer.