Copper-transporting P-Type Adenosine Triphosphatase (ATP7B) Is Expressed in Human Breast Carcinoma

This is the first report to show that a copper-transporting P-type adenosine triphosphatase, ATP7B, is expressed in certain breast carcinomas, and a priori knowledge of its expression is important for the choice of therapy. We investigated the hypothesis that ATP7B, which was shown to be associated with cisplatin resistance in vitro , is expressed in certain breast carcinomas. To test this hypothesis, ATP7B expression and protein level were examined in 41 breast carcinomas using RT-PCR and immunohistochemistry. ATP7B gene/protein could be detected in 22.0% (9/41) of breast carcinomas and ATP7B gene expression was correlated well with the protein expression. In nine ATP7B-positive tumors, adjacent normal breast tissue was similarly analyzed, revealing that ATP7B is upregulated in breast carcinoma. ATP7B gene expression in poorly differentiated carcinoma was significantly higher than that in well-/moderately differentiated carcinoma ( P =0.012). Furthermore, we found no association between the ATP7B gene/protein expression and that of MDR1, MRP1, LRP and BCRP. These findings suggested that ATP7B gene expression might be a chemoresistance marker for cisplatin in patients with poorly differentiated breast carcinoma.     

葡萄糖转运蛋白1在乳腺癌组织中的表达及与其预后的关系

目的探讨葡萄糖转运蛋白1（glucose transporter 1,GLUT-1）在乳腺癌组织中的表达及与其预后的关系。方法采用免疫组织化学SABC法,检测81例乳腺癌及20例乳腺纤维瘤组织中GLUT-1表达情况;观察GLUT-1表达与乳腺癌组织临床病理因素及预后的关系。结果乳腺癌中GLUT-1阳性表达率为61.7%（50/81）,而乳腺纤维瘤组织中未检测到GLUT-1阳性表达（P〈0.001）。GLUT-1表达与淋巴结转移数目及肿瘤有无复发、转移显著相关（P〈0.05）,而与雌孕激素受体表达和肿瘤大小无显著相关性（P〉0.05）。GLUT-1阳性组和阴性组无复发生存时间分别为（48.56±3.35）个月和（66.79±4.50）个月,具有统计学差异（P〈0.05）。结论 GLUT-1高表达提示肿瘤具有较强浸润生物学行为,GLUT-1表达与乳腺癌的预后密切相关。     

可溶性耐药相关钙结合蛋白在乳腺癌组织中的表达及其与预后关系

目的:探讨可溶性耐药相关钙结合蛋白(Soluble resistance-related calcium binding protein,Sorcin)在乳腺癌组织中的表达,评估其在乳腺癌预后中的作用。方法:采用免疫组织化学方法(IHC)检测60例手术切除的乳腺癌组织中Sorcin的表达,并分析其与临床、病理特征的关系及对预后的影响。结果:1)Sorcin在乳腺癌组织中的阳性表达率为56.7%(34/60);2)Sorcin与月经状况、肿瘤大小、淋巴结转移、组织分级和激素受体状况均无明显差异(P>0.05);3)Kaplan-Meier生存分析结果表明Sorcin表达与无瘤生存期和总生存期均无明显差异(P>0.05);4)COX单因素分析和多因素分析显示肿瘤大小、淋巴结转移和ER与无瘤生存期及总生存期(P<0.05)明显相关,PR与总生存期(P<0.05)明显相关。结论:Sorcin在乳腺癌组织中具有高表达,与乳腺癌的无瘤生存期和总生存期皆无明显差异。     

Clinical Significance of Glucose Transporter 1 (GLUT1) Expression in Human Breast Carcinoma

Glucose uptake and glycolytic metabolism are enhanced in cancer cells compared to normal cells and tissues. Increased expression of glucose transporter 1 (GLUT1) has been reported in human malignant cells. The aim of this study is to determine the expression of the facilitative glucose transporter protein GLUT1 in human breast carcinomas and a possible correlation between GLUT1 expression and clinical outcome including disease-free or overall survival. One hundred consecutive formalin-fixed, paraffin-embedded sections of invasive breast carcinomas were evaluated by means of immunohistochemical staining of GLUT1. Forty-seven (47%) of 100 breast carcinomas showed positive staining for GLUT1. Expression of GLUT1 correlated significantly with nuclear grade (P<0.001), estrogen receptor status (P=0.002), and progesterone receptor status (P=0.001). The mean disease-free survival periods of GLUTl-positive and -negative patients were 47±2.4 months and 54.3±1.3 months, respectively (P=0.017). The mean overall survival periods of GLUTl-positive and -negative patients were 48.7±2.2 and 56.1±1.3 months, respectively (P=0.043). In the multivariate analysis, disease-free survival correlated significantly with GLUT1, tumor size, and lymph node involvement (P=0.043, P=0.014, and P=0.045, respectively). In analysis of overall survival, however, lymph node involvement, tumor size, and nuclear grade were statistically significant (P=0.024, P=0.023, and P=0.003, respectively). Our data suggest that absence of GLUT1 expression significantly increases disease-free survival. These findings demonstrate that GLUT1 expression in breast carcinoma can be a marker of aggressive biological behavior and identifies a worse prognosis in breast carcinoma patients.     

Expression of CD 133 in Invasive Ductal Carcinoma of Breast

Background: CD133 is a commonly used cancer stem cell (CSC) marker in breast cancer. However, the association between CD133 expression, with clinicopathological features and prognosis in breast cancer, is poorly understood in the Indian subcontinent. This study was designed to explore the expression of CD 133 in breast carcinoma and to know its association  between CD133 and clinicopathological characteristics. Methods: A total of fifty seven cases were included in the study. All the clinicopathological parameters were collected from Department of Pathology archives. Slides, blocks, clinical information, tumor size and axillary lymph node status were obtained from medical records and the pathology reports. Immunohistochemistry was done using CD 133 antibodies. Both Cytoplasmic and membranous staining was taken a positive. Scoring was done based on percentage of positive cells and intensity of staining. MS Excel, SPSS version 22 (IBM SPSS Statistics, Somers NY, USA) was used to analyze data.p value < 0.05 was considered as statistically significant. Results: Statistically significant association between the CD 133 expression and nodal metastasis, tumor stage and Nottingham prognostic index was analysed. There was no statistical correlation between CD 133 expression age, tumor grade and tumor size. The disease free survival showed the mean disease free survival of CD 133 positivity cases was 16months. And the patients who were negative for CD 133 expression had mean survival of 30 months. By the Kaplan Mayer graph it was evident that the more the CD 133 expression the lesser was the disease free survival of the patients. Conclusion: CD 133 expression was seen in 77.08% cases and was associated with tumor stage, lymph node metastasis, poor Nottingham prognostic index and  worse disease free survival. An increasing trend of association was seen between CD 133 expression and Age, Tumor Size and Tumor grade.     

HER3在乳腺癌中的表达及临床意义

目的探讨乳腺癌患者HER3与HER2、ER和PR之间的关系及其对乳腺癌预后的影响。方法采用免疫组织化学SP法检测癌组织中HER3、HER2、ER和PR蛋白的表达,对患者随访,随访终点为总生存时间(overall survival,OS)。结果 HER2、HER3、ER和PR在136例乳腺癌患者中的阳性表达率分别为:50%、41.18%、60.9%和27.21%。HER3与年龄、淋巴结转移、分期、肿瘤类型以及化疗状态的差异无统计学意义(P>0.05)。HER3阳性与阴性患者OS差异有统计学意义(P=0.015)。HER3(+)HER2(+)组乳腺癌患者较其他组OS明显缩短(P=0.047)。HER3的表达与PR表达无明显关系(P=0.214),但可抑制ER产生(P=0.001)。HER2阳性和HER3的过度表达高度相关(P=0.000)。结论HER3可作为独立的临床预后因子,HER3阴性患者OS较长。     

乳腺癌组织中骨桥蛋白和骨连接蛋白的表达及其临床病理意义

目的探讨乳腺癌组织中骨桥蛋白(OPN)、骨连接蛋白(ON)的表达及其临床病理意义。方法采用免疫组织化学SP法检测61例浸润性乳腺癌OPN、ON的表达,并与临床病理特征及钼靶X线征象进行对比分析。结果本组乳腺癌组织中OPN和ON的阳性高表达率分别为72.13%和77.05%,均明显高于乳腺良性肿瘤(44.83%和37.93%,P<0.05)。钼靶X线发现微钙化的钙化组乳腺癌病例中OPN、ON高表达率分别为87.50%和91.67%,明显高于无钙化组(62.16%和67.57%,P<0.05)。(P>0.05)。结论OPN、ON在乳腺癌组织中高表达,OPN、ON高表达与钼靶X线微钙化征象密切相关,但与各临床病理特征及X线片中毛刺征无相关性。     

SiRNA逆转T24/ADM细胞耐药性的研究

目的探讨siRNA（small interfering RNA）对膀胱癌多药耐药性的逆转作用。方法设计针对MDR1基因的siRNA,转染膀胱癌T24/ADM细胞,应用RT-PCR分析MDR1 mRNA的表达,免疫印迹检测MDR1蛋白表达,流式细胞仪检测细胞内阿霉素积累量。MTT法检测阿霉素对T24/ADM细胞的半数抑制浓度（IC50）。结果 3条siRNA均能不同程度地逆转T24/ADM细胞的多药耐药性,使MDR1基因的mRNA及蛋白表达水平下调,细胞内阿霉素积累量显著增加（P〈0.05）。结论 siRNA可逆转膀胱癌的多药耐药性。     

Estrone and 17β-Estradiol Reverse Breast Cancer Resistance Protein-mediated Multidrug Resistance

Breast cancer resistance protein (BCRP), an adenosine triphosphate-binding cassette transporter, confers resistance to a series of anticancer reagents, including mitoxantrone, SN-38 and topotecan. In the present study, we found that estrone and l7β-estradiol potentiated the cytotoxicity of mitoxantrone, SN-38 and topotecan in BCRP-transduced K562 cells (K562/BCRP). These estrogens showed only a marginal effect, or none, in parental K562 cells. Estrone and 17β-estradiol increased the cellular accumulation of topotecan in K562/BCRP cells, but not in K562 cells, suggesting that these estrogens inhibit the BCRP-mediated drug efflux and overcome drug resistance.     

C-erbB-2基因扩增/表达和Cathepsin-D表达对乳腺癌预后的影响

用差异ＰＣＲ方法检测了１３１例乳腺癌Ｃ－ｅｒｂＢ－２基因扩增，免疫组化法检测了１３１例乳腺癌Ｃ－ｅｒｂＢ－２和Ｃａｔｈｅｐｓｉｎ－Ｄ表达，并对其对乳腺癌的预后影响进行评价。Ｃ－ｅｒｂＢ２基因扩增阳性率为２９．０１％，与临床分期、病理类型、组织学分级和淋巴结转移状况无明显相关，与Ｃ－ｅｒｂＢ－２蛋白表达虽不完全吻合但呈高度相关。Ｃ－ｅｒｂＢ－２表达阳性率为３２．０６％，与临床分期、病理类型、组织学分级、淋巴结转移状况及瘤体大小未见明显相关，转移灶和原发灶表达一致。Ｃａｔｈｅｐ－ｓｉｎ－Ｄ表达阳性率为３２．０６％，与淋巴瘤结转移状况、Ｃ－ｅｒｂＢ－２表达及病理类型明显相关。单因素分析表明Ｃ－ｅｒｂＢ－２基因扩增、Ｃ－ｅｒｂＢ－２表达及Ｃａｔｈ－Ｄ表达对乳腺癌术后生存及无瘤生存影响显著，Ｃｏｘ模型多因素回归分析表明Ｃ－ｅｒｂＢ－２表达是乳腺癌一独立预后指标。联合应用Ｃ－ｅｒｂＢ－２和Ｃａｔｈ－Ｄ表达检测更有助于早期病例和淋巴结转移阴性病例中高危患者的筛选。     

乳腺癌组织中Ang-2及VEGF的表达及意义

目的研究乳腺癌组织中血管生成素(Ang-2)和血管内皮细胞生长因子(VEGF)的表达及临床意义。方法采用免疫组织化学SABC法,检测85例乳腺癌及42例乳腺纤维腺瘤组织中Ang-2及VEGF的表达情况,并评价两指标与乳腺癌临床病理学因素之间的关系。结果乳腺癌组织中Ang-2阳性率(68.24%,58/85)明显高于乳腺纤维腺瘤(7.14%,3/42)(P<0.0001),乳腺癌组织中VEGF阳性率(57.65%,49/85)也明显高于乳腺纤维腺瘤(4.76%,2/42)(P<0.0001)。Ang-2和VEGF表达与患者年龄及肿瘤大小无明显关系(P>0.05)。Ⅲ、Ⅳ期乳腺癌和淋巴结转移阳性组的VEGF表达阳性率明显高于Ⅰ、Ⅱ期乳腺癌和淋巴结转移阴性组(P<0.0001)。Ⅲ、Ⅳ期乳腺癌组的Ang-2表达阳性率高于Ⅰ、Ⅱ期乳腺癌组(P=0.015)。淋巴结转移阳性组的Ang-2阳性率明显高于淋巴结转移阴性组(P<0.0001)。Ang-2和VEGF两指标的表达具有相关性(P<0.05)。结论乳腺癌组织中Ang-2及VEGF有异常表达,与乳腺癌的预后有关。     

p-糖蛋白(pgp)在乳腺癌组织中的表达及与预后的关系

目的：研究P－糖蛋白（P－glycoprotein,Pgp)在乳腺癌组织中的表达,评估其在乳腺癌预后中的作用。方法：采用免疫组织化学方法（Immunohistochemistry,IHC)检测47例手术切除的乳腺癌组织中Pgp的表达,并分析其与临床、病理特征的关系及对预的的影响。结果：Pgp在乳腺癌组织中的总表达率为83．0％（39／47例）,高表达者占53．2％（25／47例）,其与月经状、肿瘤大小、淋巴结转移、组织分级和激素受体状况均无关（P＞0．05）;Kaplan-Meier生存分析结果表明Pgp表达与无病生存期和总生存期均显著相关（P＜0．01）,而多因素Cox分析却显著仅有Pgp和淋巴结转移是独立的预后因素,结论：Pgp在乳腺癌组织中具有较高的表达,有可能成为判断乳腺癌预后的有用指标。     

岩藻糖转移酶9在乳腺癌中的表达及其临床意义

目的 本研究探讨岩藻糖转移酶9（fucosyltransferase9,FUT9）在乳腺癌组织中的表达及其与乳腺癌临床病理因素的关系。方法 采用免疫组织化学SP法检测11例良性乳腺增生组织和114例乳腺癌组织（其中47例合并转移淋巴结,67例无转移淋巴结）中FUT9的表达。结果 FUT9在良性乳腺增生组织中的阳性表达率为9.1%（1/11）,而在乳腺癌组织中的阳性表达率为71.9%(82/114),在转移的淋巴灶中阳性表达率为100%（47/47）。乳腺癌组织中的阳性表达率明显高于良性乳腺增生组织的阳性表达率,两组比较差异有统计学意义（P<0.01）。乳腺癌组织中有淋巴结转移组的阳性表达率为80.9%（38/47）,明显高于无淋巴结转移组的阳性表达率65.7%（44/67）,两组比较差异有统计学意义（P<0.05）。在有淋巴结转移组FUT9的表达与TNM分期以及转移的淋巴结个数相关（P<0.05）。FUT9的表达与ER、PR、HER2的表达无相关性（P>0.05）。结论 FUT9在乳腺癌组织中的表达明显增高;FUT9的表达与淋巴结转移、转移的淋巴结个数以及TNM分期有关;FUT9可能与乳腺癌淋巴道转移有关。     

DcR3对乳腺癌细胞凋亡的影响及其在乳腺癌血清中的表达

目的研究诱捕受体3（DcR3）单克隆中和性抗体对乳腺癌细胞生长和凋亡的影响,以及DcR3在乳腺癌患者血清中的表达及临床意义。方法将DcR3单克隆中和性抗体作用于体外培养的人乳腺癌细胞系MCF-7,检测细胞增殖情况及Caspase 3/7活性变化,应用Hoechst 33342及碘化丙啶双重染色荧光显微镜下观察细胞凋亡形态及计算凋亡率。应用ELISA检测乳腺癌患者血清中DcR3的表达。结果DcR3中和性抗体可抑制MCF-7细胞生长并可诱导细胞凋亡（P<0.05）。乳腺癌患者血清DcR3 水平明显高于健康对照组（P<0.01）;DcR3表达水平与肿瘤大小及临床TNM分期有关（P<0.05）。结论DcR3对人乳腺癌细胞增殖具有抑制作用,其机制与诱导肿瘤细胞凋亡有关。DcR3与乳腺癌的发生及恶性进展有关,检测DcR3血清表达有助于乳腺癌诊断及预后判断。     

IL-8在乳腺癌组织中的表达及其临床意义

目的:检测乳腺癌组织中IL-8表达,探讨其与临床病理学因子及生物学因子的关系.方法:采用免疫组化SP法检测乳腺癌组织中IL-8、ER、PR、VEGF、MVD表达.结果:1)103例乳腺癌组织中IL-8( )表达率为34%、( )为35%、( )为10.7%,与正常及良性组织相比有显著性差异(P=0);2)IL-8表达水平与VEGF、MVD表达呈正相关(P=0);3)IL-8表达与淋巴结转移状况、组织学分级、临床分期有关(P＜0.05);4)IL-8与乳腺癌预后有关(P＜0.05);5)IL-8与ER、PR、肿瘤大小及病理类型无关(P＞0.05).结论:IL-8的表达水平与影响乳腺癌患者预后的生物学因素相关,IL-8高表达,预后差,可作为判断乳腺癌预后的指标.     

Ku80蛋白在乳腺癌组织中的表达及其临床意义

目的:研究Ku80蛋白在乳腺癌的表达及与临床病理特征的关系.方法:采用免疫组化法检测65例乳腺癌组织Ku80、ER、PR、HER-2和突变型p53表达水平.结果:65例乳腺癌标本中Ku80、ER、PR、HER-2、p53阳性率分别为78.5%、40.0%、53.8%、44.6%和32.3%.Ku80表达水平与肿瘤大小、ER、PR、HER-2和p53的表达水平有关(P＜0.05),其中Ku80与HER-2密切相关(rs=0.319,P=0.010).结论:Ku80表达水平与影响乳腺癌患者预后的生物学特征相关,特别是与HER-2关系密切;Ku80与HER-2之间可能存在调控通路.     

Immunohistochemical Expression of Blood Group Substances and Related Carbohydrate Antigens in Breast Carcinoma

In forty–one carcinomas and sixteen benign lesions (fibroadenoma and mastopathy) of the human breast, immunohistochemical expression of sialylated and non–sialylated forms of both Le^a and Le^x, and the A, B, and H type 2 blood group substances were studied by using an indirect immunoperoxidase staining. In normal ductal epithelium and benign lesion of breast, Lewis–related antigens were mostly expressed. Breast carcinomas showed these antigens with the following frequencies: Le^a, 31.7% (13/41); sialyl Le^a, 56.1% (23/41); Le^x, 46.3% (19/41); sialyl Le^x, 68.3% (28/41); A/B/H type 2, 38.1% (16/41). Sialylated forms of Le^a and Le^x were observed more frequently than their respective non–sialylated forms in breast carcinomas. In both one normal epithelium and four carcinomas of breast with Le^(a?b-) phenotype, the expressions of type 2 antigens were observed, while type 1 antigens were not consistently expressed. Although compatible expression was observed in all specimens of both normal epithelium and benign lesion of breast, twenty–four cases with the deletion of A and/or B antigens, six cases with H type 2 accumulation and one case with incompatible expression were demonstrated in breast carcinoma. Thirty–one breast carcinomas which showed the deletion of A/B/H type 2 expressed the Lewis–related antigens more frequently than nine cases which showed compatible expression. These results suggested that the activation of terminal fucosyltransferase and sialyltransferase as well as inactivation of some glycosyltransferases had occurred in cancer cell membrane, and sialyl Le¹, defined by a new monoclonal antibody CSLEX1, may be useful as a tumor–associated antigen independent of Lewis blood group type in breast cancer.     

环氧化酶-2在乳腺浸润癌中的表达及临床意义

 目的　研究环氧化酶 2 (COX 2 )在乳腺浸润癌中的表达情况及其与乳腺癌临床病理特征的关系。方法　应用免疫组织化学 (SP法 )检测 83例乳腺浸润癌 (其中有周围组织浸润 6 2例 ,伴腋淋巴结转移 4 7例 )和 35例癌旁组织中COX 2的表达。结果　COX 2在乳腺浸润癌中的阳性表达率为 73.5 %(6 1/ 83) ,显著高于癌旁组织 (P <0 .0 0 1) ;乳腺浸润癌组织COX 2的表达与临床分期、周围组织浸润和淋巴结转移密切相关 ,(P <0 .0 5 ,P <0 .0 5 ,P <0 .0 0 1) ,与患者年龄和肿瘤大小无明显相关性。结论　COX 2蛋白表达可能在乳腺浸润癌的发生、发展中起重要作用。     

乳腺癌组织中Hedgehog相关蛋白的表达及其临床意义

目的 探讨Hedgehog相关蛋白在乳腺癌中的表达及其临床意义。方法 从334例乳腺癌 组织中制备组织块。免疫组织化学法检测六种Hedgehog信号蛋白（Shh、ptch、Smo、Gli-1、 Gli-2和Gli-3）的表达。结果 Hedgehog信号蛋白表达与一些预后因素显著相关,包括淋巴结转 移与Shh（P=0.001）、Gli-3 (P<0.001)、Ptch(P=0.025)的相关性、分期与Shh和Ptch表达（P分别 为0.007和0.037）的相关性,核分级与Smo(P=0.004)和Gli-3（P=0.026）的相关性,组织学分级与 Smo(P=0.007)的相关性。Shh、Ptch、Smo、Gli-1、Gli-2和Gli-3表达在表型之间差异有统计学意义 （P分别为<0.0001、<0.0001、0.004、0.039、0.031和0.003）。Gli-2表达与较差的整体生存结果相关 （P=0.012）。结论 Hedgehog通路表达激活与乳腺癌的侵袭性相关,可能是乳腺癌的一个有效的治 疗靶点。Gli-2可作为乳腺癌的一个预后指标。     

槲皮素联合奥曲肽对人乳腺癌细胞株MDA-MB-231增殖的影响

目的: 研究槲皮素(quercetin,QUE)联合奥曲肽(octreotide,OCT)对人乳腺癌细胞增殖的影响. 方法: 1)以QUE联合OCT在5个不同浓度水平分别作用于体外培养的人乳腺癌细胞株MDA-MB-231,四甲基偶氮唑盐(methyl thiazolyl tetrazolium,MTT)法测定细胞光密度(optical density,OD).2)以QUE与OCT联用时半数抑制浓度(50%inhibited concentration,IC50)的1/2作用于体外培养的人乳腺癌细胞株MDA-MB-231,流式细胞仪检测细胞周期分布,免疫细胞化学分析端粒酶的表达. 结果: QUE与OCT在不同浓度联用于MDA-MB-231细胞,其抑制率(fa)分别是1.45%、3.63%、8.32%、13.41%、85.78%,合用指数(Combination index,CI)均<1.两药联用能阻滞细胞周期于G0～G1期(P<0.01),下调端粒酶表达(P<0.01). 结论: QUE联合OCT能协同抑制MDA-MB-231细胞增殖,其机制与阻滞细胞周期、抑制端粒酶活性有关.