胃癌组织中Ets-1的表达及其与血管生成临床病理特征和预后的关系

目的 探讨胃癌组织、癌旁组织及淋巴结转移灶中Ets-1表达的临床意义,分析Ets-1表达与血管生成、临床病理特征及预后的关系.方法 应用免疫组化SP法,采用组织芯片技术,检测189例胃癌组织、54例癌旁组织、41例淋巴结转移灶及32例正常胃黏膜中Ets-1蛋白的表达.对胃癌患者通过上门或电话进行问卷随访.结果 胃癌组织、癌旁组织和正常胃黏膜Ets-1的阳性表达率分别为71.4%、29.6%和18.8%,3组间差异有统计学意义(P＜0.01).135例Ets-1表达阳性的胃癌组织微血管密度(MVD)为30.42±15.21,54例Ets-1表达阴性的胃癌组织MVD为25.73±11.50.两组差异有统计学意义(P=0.042).Ets-1蛋白表达与浸润深度、淋巴结转移有关(P＜0.01),与性别、年龄、肿瘤大小、分化程度、Lanren分型无关(P＞0.05).41例淋巴结转移灶和相对应的41例胃癌组织Ets-1表达阳性率分别为84.4%和58.5%,差异有统计学意义(P=0.007).单因素分析显示,Ets-1表达对胃癌患者生存期的影响有统计学意义(P＜0.05),Cox多元回归分析显示,Ets-1表达不是胃癌患者预后的独立影响因素(P＞0.05).结论 Ets-1在促进胃癌血管生成中发挥着重要作用,在胃癌的发生、发展中扮演了重要角色,Ets-1表达对胃癌患者的生存期有一定影响,胃癌淋巴结转移灶和原发灶肿瘤组织中Ets-1的表达不同,具有异质性.     

KAI1 protein is down-regulated during the progression of human breast cancer.

The KAI1 gene was identified as a metastasis suppressor gene for human prostate cancer. Recently, we showed that KAI1 mRNA levels were higher in an immortal, normal-like breast epithelial cell line and nonmetastatic breast cancer cell lines but lower substantially in highly metastatic breast cancer cell lines. In this study, we examined KAI1 protein expression in breast cancer cell lines by Western blot and immunohistochemical study. KAI1 protein levels paralleled KAI1 mRNA levels and were inversely correlated with the metastatic potential of breast cancer cells. Furthermore, we examined KAI1 protein expression immunohistochemically in specimens from 81 patients with breast cancer and then correlated the findings with the clinical and histopathological parameters of the patients. High levels of KAI1 protein expression were found in normal breast tissues and noninvasive breast cancer (ductal carcinoma in situ). In contrast, KAI1 expression was reduced in most of the infiltrating breast tumors. We found that, in general, more malignant tumors demonstrated significantly lower KAI1 expression (P = 0.004). Additionally, among 29 specimens demonstrating multiple stages of malignancy within a single specimen, 23 demonstrated significant differences in KAI1 expression between benign breast tissue, ductal carcinoma in situ, and invasive carcinoma. The higher the incidence for malignancy within a given specimen, the lower the KAI1 expression (P < 0.001). These data suggest that in advanced breast cancer, KAI1 expression is down-regulated. Therefore, KAI1 may be a potentially useful indicator of human breast cancer progression.     

人内源性逆转录病毒K 包膜蛋白在乳腺肿瘤组织中的表达及意义*

目的：检测人内源性逆转录病毒K（HERV-K ）包膜蛋白（env 蛋白）在不同乳腺肿瘤组织中的表达,分析其表达水平与乳腺癌患者临床病理特征的相关性,探讨HERV-K env蛋白的表达与乳腺癌预后的关系。方法：免疫组织化学法检测HERV-K env 蛋白在95例乳腺癌组织、30例乳腺良性肿瘤组织中的表达情况,χ2检验用于分析HERV-K env蛋白的表达水平与乳腺癌患者临床病理特征的相关性。结果：免疫组织化学结果显示,HERV-K env蛋白的阳性表达主要位于细胞浆,该蛋白在乳腺癌组织中多呈强阳性或中度阳性表达,而在乳腺良性肿瘤组织中多呈阴性或弱阳性表达。HERV-K env蛋白在乳腺癌组织中的阳性表达率（88.4%）明显高于乳腺良性肿瘤组织（40.0%）,差异具有统计学意义（P<0.001）。 HERV-K env蛋白的表达水平与乳腺癌的某些预后因素存在相关性。在HERV-K env蛋白表达阳性的乳腺癌患者中,该蛋白的表达水平与肿瘤大小、临床分期、腋窝淋巴结转移数目之间存在正相关关系,与ER的表达状况之间存在负相关关系（P 均<0.05）,而与患者的年龄、组织学分级以及PR的表达状况无相关性（P 均>0.05）。 结论：HERV-K env蛋白在乳腺良性肿瘤中多为阴性表达,而在乳腺癌中多呈阳性表达,并且可能与乳腺癌的发展及预后相关,该蛋白在乳腺癌中的表达水平可能是一种判断乳腺癌预后的指标。     

乳腺癌组织中缺氧诱导因子-1α和葡萄糖转运蛋白1表达及其意义

目的探讨乳腺癌组织中缺氧诱导因子-1α（HIF-1α）和葡萄糖转运蛋白1（Glut1）表达的关联性及其与细胞增生活性和临床病理因素的关系。方法采用免疫组织化学方法检测HIF-1α、Glut1、增生细胞核抗原（PCNA）在80例乳腺癌组织、20例乳腺纤维腺瘤及20例乳腺增生组织中的表达水平并比较它们之间的相关性。结果HIF-1α和Glut1在乳腺纤维腺瘤、乳腺增生组织中不表达；HIF-1α在乳腺导管内原位癌中阳性率55．0％（11／20），浸润性乳腺癌中85．0％（51／60）；乳腺癌中Glut1阳性率58．8％（47／80）；乳腺癌中PCNA阳性率75％（60／80），其中原位癌65％（13／20），浸润癌78．3％（47／60）。乳腺癌中HIF-1α表达与Glut1呈显著正相关（r=0．653，P〈0．01）；HIF-1α表达与PCNA呈显著正相关（r=0．693，P〈0．01）；Glut1与PCNA呈显著正性相关（r=0．742，P〈0．01）。HIF-1α和Glut1在淋巴结、雌激素受体状态及组织学分级中表达差异有统计学意义；Glut1在肿瘤大小及孕激素受体状态表达差异也有统计学意义。结论HIF-1α和Glut1蛋白的过表达共同参与了乳腺癌的发生、发展，与乳腺癌细胞增生活性密切相关，有望成为乳腺癌治疗的新靶点。     

Loss of SFRP1 is associated with breast cancer progression and poor prognosis in early stage tumors

Aberrant activation of the Wnt signaling pathway plays an important role in the development of solid tumors such as breast and colon cancer. Secreted Frizzled-related protein 1 (SFRP1) is a negative regulator of the Wnt pathway. It has been described that SFRP1 mRNA is strongly down-regulated in breast cancer and a putative tumor suppressor function has been postulated. We have generated and characterized an SFRP1 specific antibody to analyze its expression on protein level and to investigate the association of SFRP1 expression with clinicopathological parameters and patient survival. Analysis of >2000 invasive breast tumors and 56 carcinoma in situ revealed similar frequencies of SFRP1 loss in these tumors (46% and 43% respectively). Therefore, we propose that loss of SFRP1 expression is an early event in breast tumorigenesis. SFRP1 expression was inversely correlated with tumor stage (p<0.001) but not with tumor grade (p=0.14) or lymph node status (p=0.84). Performing a multivariate analysis we could confirm the association between tumor stage and SFRP1 expression (p=0.029). In particular, loss of SFRP1 expression in early stage breast tumors (pT1) was associated with poor prognosis (p=0.04). In conclusion, expression of SFRP1 is commonly lost in breast cancer. SFRP1 expression might be useful as a novel prognostic marker in early stage breast cancer.     

Bmi-1在乳腺癌组织中的表达及意义

背景与目的:多梳基因家族的Bmi-1被认为是一种癌基因,在多种肿瘤组织中均有表达,但其与乳腺癌关系的研究国内尚无报道.本研究旨在通过检测中国人乳腺癌组织中Bmi-1的表达,探讨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58例乳腺癌组织中Bmi-1蛋白的表达情况,并结合患者的临床病理资料分析与其相关性.结果:所检测的58例乳腺癌标本中Bmi-1蛋白强阳性表达48例(82.8%).统计分析结果发现Bmi-1的强阳性表达与乳腺癌的淋巴结转移及临床分期密切相关(P＜0.05),而与肿瘤大小及ER、PR、cerbB-2、VEGF等临床病理特征无显著性相关(P＞0.05).结论:Bmi-1蛋白在乳腺癌组织中的高表达与肿瘤进展相关,预示肿瘤具有高度转移潜能;Bmi-1可能成为预测乳腺癌高度转移的新分子标志物.     

乳腺癌 nm23-H_1 蛋白表达的研究

应用ＳＰ免疫组化法检测８４例乳腺浸润性导管癌中ｎｍ２３－Ｈ１蛋白的表达，结果显示ｎｍ２３－Ｈ１蛋白在７２例（８７．５％）有表达。研究表明ｎｍ２３－Ｈ１蛋白的表达水平与淋巴结转移呈负相关（Ｐ＜０．０１），与ＥＲ、ＰＲ水平呈正相关（Ｐ＜０．０１，Ｐ＜０．０５），而与组织学分级ＰＣＮＡ指数及ＭＤＲ１－Ｐｇｐ的表达无关（Ｐ＞０．０５）。     

HYAL1 overexpression is correlated with the malignant behavior of human breast cancer

Extracellular matrix (ECM) is closely correlated with tumor cell growth, proliferation, metastasis and angiogenesis, etc. Hyaluronic acid (HA) is a component of the ECM, and hyaluronidase (HAase) is a HA‐degrading endoglycosidase. Levels of HAase are elevated in many cancers. Hyaluronidase‐1 (HYAL1) is the major tumor‐derived HAase. In this study, we detected HYAL1 expression levels in breast cancer cells and tissues, and measured the amount HAase activity in breast cancer cells. Compared with nonmalignant breast cell line HBL‐100 and normal breast tissues, HYAL1 were overexpressed in breast cancer cell lines MDA‐MB‐231, MCF‐7, invasive duct cancer tissues and metastatic lymph nodes, respectively. Accordingly, the amount HAase activity in MDA‐MB‐231 and MCF‐7 was higher than that in HBL‐100. In addition, knockdown of HYAL1 expression in MDA‐MB‐231 and MCF‐7 cells resulted in decreased cell growth, adhesion, invasion and angiogenesis potential. Meantime, the HYAL1 knockdown markedly inhibited breast cancer cell xenograft tumor growth and microvessel density. Further studies showed that the HYAL1, HYAL2 and HA were elevated in breast cancer, and HYAL1 could downregulate HA expression. In conclusion, HYAL1 may be a potential prognostic marker and therapeutic target in breast cancer.     

plexin-B1 在乳腺癌组织中的表达及其与淋巴道转移的关系

 目的 探讨plexin-B1在乳腺癌组织中的表达及其与乳腺癌侵袭、转移的关系。方法 应用免疫组化supervision法分别检测15例正常乳腺组织和85例乳腺癌组织中plexin-B1的表达。结果 plexin-B1在正常乳腺组织中均为阴性表达，在乳腺癌组织中癌细胞和管腔内皮细胞的表达阳性率分别为76．5％（65／85）和70．6％（60／85），其中在有淋巴结转移组分别为87．0％（47／54）和85．2％（46／54），在无淋巴结转移组分别为58．1％（18／31）和45．2％（14／31），两组间的差异有统计学意义（P〈0.001）。plexin-B1在乳腺癌组织中表达与雌激素受体（estrogenreceptor，ER）的状态高度相关，在肿瘤细胞与淋巴管内皮细胞的表达呈显著的正相关（r=0．666，P〈0．01））。但其表达与肿瘤大小、临床分期及淋巴结转移数目无关。结论 乳腺癌组织中plexin-B1的高表达可能与乳腺癌发生及淋巴道转移密切相关。     

l‐type amino acid transporter 1 and CD98 expression in primary and metastatic sites of human neoplasms

The significance of l ‐type amino acid transporter (LAT) 1 expression remains unclear in the metastatic process of human neoplasms, whereas experimental studies have demonstrated that LAT1 is associated with the metastatic process of cancer cells. We compared the immunohistochemical expression of LAT1 and CD98 between the primary site and a concordant pulmonary metastatic site in 93 cancer patients, all of whom had undergone thoracotomy. LAT1, CD98, Ki‐67 labeling index, vascular endothelial growth factor (VEGF), CD31, and CD34 were analyzed by immunohistochemical staining in the resected tumors of 93 cancer patients: 45 colon cancers; nine breast cancers; eight head and neck cancers; 11 genital cancers; 14 soft‐tissue sarcomas; and six other cancers. The expression of these markers was significantly higher in the metastatic sites than in the primary sites. In total, the positive rates of LAT1, CD98, Ki‐67, VEGF, CD31, and CD34 were 40, 24, 56, 41, 45, and 39%, respectively, in the primary sites and 65, 45, 84, 67, 73, and 61%, respectively, in the metastatic sites. LAT1 expression was closely correlated with CD98 expression, angiogenesis, and cell proliferation. The association between LAT1 and CD98 expression was strongest in the primary and metastatic sites. The present study suggests that overexpression of LAT1 and CD98 has an important role to play in the metastatic process of variable human neoplasms. Moreover, LAT1 expression was significantly correlated with cell proliferation and angiogenesis. (Cancer Sci 2008; 99: 2380–2386)     

Role of ICAM1 in invasion of human breast cancer cells

We identified previously a region on chromosome 19p13.2 spanning the genes encoding the intercellular adhesion molecules (ICAM), ICAM1, ICAM4 and ICAM5 as a breast cancer susceptibility locus. Genetic variants in this region were also associated with indicators of disease severity, including higher rates of metastases to other organs. Based on this association, we set out to explore the role of ICAM1 in proliferation and invasion of human breast cancer cells. We observed that ICAM1 downregulation at the mRNA and protein levels led to a strong suppression of human breast cell invasion through a matrigel matrix. Under the same conditions, no significant effect on cell proliferation in vitro was seen. Incubation of cells with an antibody against ICAM1 blocked invasion of the highly metastatic MDA-MB-435 cell line in a dose-dependent manner without affecting cell migration. We also demonstrated that the level of ICAM1 protein expression on the cell surface positively correlated with metastatic potential of five human breast cancer cell lines and that ICAM1 mRNA levels were elevated in breast tumor compared with adjacent normal tissue. These results corroborate our previous genetic finding that variations in the ICAM region are associated with the occurrence of metastases and establish a causal role of ICAM1 in invasion of metastatic human breast carcinoma cell lines.     

Jun activation domain-binding protein 1 expression in breast cancer inversely correlates with the cell cycle inhibitor p27(Kip1)

The Jun activation domain-binding protein 1 (JAB1), aside from being an activator protein 1 coactivator, is involved in degradation of the cyclin-dependent kinase inhibitor p27. We examined JAB1 and p27 protein expression in invasive breast carcinoma specimens and the association of this expression with clinical outcome. JAB1 was detected immunohistochemically in 43 of 53 (81%) tumors; 32 (60%) breast carcinomas showed high JAB1 expression (>50% of cells positive) and reduced or absent p27 levels (P = 0.02, Mann-Whitney U test). Tumors with high p27 expression were rarely positive for JAB1. All eight patients with JAB1-negative tumors had no evidence of relapse or disease progression at a median follow-up of 70 months. Immunoblotting showed strong JAB1 expression in breast carcinoma samples but not in paired normal breast epithelial samples. Targeted overexpression of JAB1 by regulated adenovirus in breast cancer cell lines also reduced p27 levels by accelerating degradation of p27. Thus, the JAB1:p27 ratio may be a novel indicator of aggressive, high-grade tumor behavior, and control of JAB1 could be a novel target for experimental therapies.     

Functional domains of CCN1 (Cyr61) regulate breast cancer progression

CCN1 plays diverse roles in cellular proliferation, survival, migration and angiogenesis. We determined the relationship between CCN1 protein expression and clinical factors that are important for the classification of breast cancer. CCN1 contains four functional domains; the contribution of each of the structural domains to the biological properties of CCN1 in breast cancer was investigated. We performed immunohistochemistry for CCN1 on a breast cancer tissue array, and conducted a detailed statistical analysis on the relationship between CCN1 protein expression and clinical factors that are important for the classification of breast cancer. The structure-function relationship was examined using four mutant constructs in which one of the modules (DM1-DM4) had been deleted. MCF-7 breast cancer cells were stably transfected with these constructs and their biological activity was tested in comparison to full-length CCN1. Staining of CCN1 in tumors was positively correlated with AJCC disease stage. A strong association also was found between lymph node involvement and high CCN1 expression in patients with invasive breast cancer; there was a significant increase in the breast cancer expression of CCN1 in patients with positive lymph nodes (P=0.004), and the levels of CCN1 correlated with the number of positive lymph nodes (P=0.0006). Deletion of module 4 rendered CCN1 unable to either bind heparin or associate with the extracellular matrix. Furthermore, MCF-7/DM4 cells demonstrated reduced cell spreading, migration and proliferation, indicating that module 4 of the protein is important for its ability to promote these activities. These findings indicate that CCN1 is involved throughout the clinical progression of breast cancer to an invasive phenotype. The multimodular structure of CCN1 enables it to fulfill multiple functions that may contribute to the different stages of cancer development, raising the prospect that specific regions of CCN1 could be targeted for therapeutic benefit to inhibit particular aspects of malignancy in breast cancer.     

Decreased microRNA-214 levels in breast cancer cells coincides with increased cell proliferation, invasion and accumulation of the Polycomb Ezh2 methyltransferase

MicroRNAs (miRNAs) are small non-coding RNAs, which regulate gene expression by inhibiting translation or promoting degradation of specific target messenger RNAs (mRNAs). Alteration of the levels of a number of miRNAs is common in solid and hematological tumors. We have shown previously that miR-214 regulates Ezh2 in skeletal muscle and embryonic stem cells. The current study was aimed at examining the role of miR-214 in breast cancer where miR-214 levels are reduced but whether this phenomenon bears a functional relevance is unknown. MiR-214 expression was inversely correlated with Ezh2 mRNA and protein levels in breast cancer cell lines and at least one copy of the miR-214 alleles was found to be deleted in 24% (6/25) of primary breast tumors. Experimental increase of miR-214 in breast cancer cell lines correlated with reduction of Ezh2 protein levels, a known marker of invasion and aggressive breast cancer behavior. Supporting a direct targeting mechanism, miR-214 decreased luciferase activity from a construct containing the Ezh2 3' untranslated region. Expression of miR-214 specifically reduced cell proliferation of breast cancer cells and inhibited the invasive potential of a highly metastatic breast cancer cell line. These findings indicate that reduced miR-214 levels may contribute to breast tumorigenesis by allowing abnormally elevated Ezh2 accumulation and subsequent unchecked cell proliferation and invasion.     

Expression of the HER family mRNA in breast cancer tissue and association with cell cycle inhibitors p21(waf1) and p27(kip1)

BACKGROUND: The HER family of the receptor tyrosine kinases epidermal growth factor receptor (EGFR), HER2, HER3 and HER4 are involved in the pathogenesis of many human malignancies. Although there is extensive literature on the expression of single HER-2 and EGFR receptors in breast cancer, little is known concerning the simultaneous expression at the mRNA level of these four receptors in human breast tissue and their influence in downstream signaling pathways that control cell cycle and proliferation. MATERIALS AND METHODS: The mRNA expression pattern of the four HER-receptors has been investigated and correlated with the expression of the cyclin-dependent kinase (CDK) inhibitors p21(Waf1) and p27(Kip1) in 67 breast cancer specimens. RESULTS: A positive correlation between HER-3 and HER-4 mRNA levels and a negative correlation between HER-2 and HER-3 was found. Compared to normal breast tissue, all four receptors were overexpressed in breast tumors and the strongest overexpression was found for HER-3 (p = 0.001). HER-4 expression was inversely related to histopathological grading (HPG), suggesting that elevated HER-4 mRNA expressions could be a biological marker of a more differentiated phenotype. The expression of p21(Waf1) protein was higher in HER-2-negative tumors, compared to HER-2-positive breast carcinomas. Compared to normal breast tissue, p21delta, the 19 kDa degraded form of p21 protein, was markedly expressed in breast cancer (p < 0.001). Conversely, p27(Kip1) was positively associated with HER-2 receptor and inversely associated with HER-3. CONCLUSION: The HER family members are overexpressed in breast cancer. Complex patterns of HER family expression were observed and the effect on cell cycle regulation was dependent on that pattern.     

Plexin-B2 在人乳腺癌中的表达及临床意义*

目的:探讨人乳腺癌中Plexin-B 2 的表达,以及其与人表皮生长因子受体-2（Her-2）共表达与乳腺癌恶性行为的关系。方法:免疫组化SP法检测15例正常乳腺组织中Plexin-B 2 蛋白的表达,及112 例乳腺癌组织中Plexin-B 2 和Her-2 蛋白的表达。结果:Plexin-B 2 在癌细胞和正常乳腺组织小叶及导管上皮细胞中的阳性表达率分别为69.64% 和60.00% ,差异无统计学意义（P>0.05）。 Plexin-B 2 在乳腺癌组织癌周微脉管中的阳性表达率为44.64% ,而正常乳腺微脉管均为阴性。癌细胞和癌周微脉管中Plexin-B 2 的表达正相关（r=0.593,P=0.000）,并且都与临床分期及淋巴结转移有关（P<0.05）。 Plexin-B 2、Her-2 共表达比Plex in-B 2 单阳性的肿瘤临床分期晚、淋巴结转移率高,差异有统计学意义（P<0.05）。 结论:Plexin-B 2 异常表达可能与乳腺癌的恶性进展有关,Plexin-B 2-Her- 2 共表达可能促使乳腺癌的侵袭转移。