Dose intensity as a therapeutic strategy in breast cancer

The results of systemic treatment for primary and metastatic breast cancer have plateaued in the past decade. The majority of oncologists continue to use the same chemotherapy regimens and endocrine therapies that were available in the mid 1970s. In metastatic breast cancer, still only 60–70% of patients can be expected to achieve a remission, with only 10–20% of these being a complete remission, which is usually of short duration. Metastatic breast cancer remains incurable today. Obviously, new treatment strategies are needed. The development of new active drugs, or the development of innovative ways of giving old drugs, has been disappointing in breast cancer. Similarly, combining hormones with chemotherapy, or the use of various biologic response modifiers, has not resulted in a major advance. One strategy that is currently undergoing active research is increased dose intensity of chemotherapy. This can be achieved by delivering extremely high doses of cytotoxic chemotherapy followed by hematopoietic support. A second approach involves delivering lower doses, but on a more frequent schedule than conventional programs. Preliminary results from phase II evaluation of these programs demonstrate high complete response rates, relatively short response durations, and considerable toxicity. However, 10–20% of patients treated with these regimens remain in complete remission several years after treatment, providing optimism that this approach may be effective in some patients. Advances in hematopoietic support, including autologous bone marrow transplantation (ABMT), peripheral stem cell administration, and the use of hematopoietic growth factors, have reduced toxicity. This strategy is now ready for phase III randomized trials in metastatic breast cancer as well as in the high-risk adjuvant patient to determine if high-dose therapy offers a worthwhile advantage in patients with breast cancer.     

Clinicopathological features and treatment strategy for triple-negative breast cancer

Breast cancers are divided into at least 4 subtypes on the basis of gene expression profiles and expression of receptors (hormone receptors (HR) and HER2) as measured by immunohistochemistry. These subtypes have different prognoses and responses to treatments such as endocrine manipulation, anti-HER2 therapy, and chemotherapy. Triple-negative breast cancer (TNBC) is immunohistochemically defined as lacking estrogen and progesterone receptors and not overexpressing HER2. TNBC accounts for approximately 15% of breast cancer patients, and is more chemosensitive but has a worse prognosis than the HR-positive/HER2-negative phenotype. TNBC is a heterogeneous disease that does not offer specific targets in the same way as HR-positive and HER2-positive breast cancers, and is similar to basal-like breast cancer and BRCA1-related breast cancer. At present, the lack of highly effective therapeutic targets for TNBC leaves standard chemotherapy, for example the combination of anthracycline and taxane, as the only medical treatment, but this is insufficiently efficacious. Novel approaches for TNBC, for example DNA damaging agents, PARP-1 inhibitors, receptor tyrosin kinase inhibitors (TKIs), and antiangiogenesis agents, have been examined in clinical settings. Concerning therapeutic strategies for TNBC, it is most important to develop novel effective approaches for TNBC treatment and high-throughput predictive tools for standard chemotherapy and novel agents.     

血清蛋白质谱诊断模型在乳腺癌辅助诊断中的价值

目的 建立乳腺癌的血清蛋白质谱诊断模型,评价其在乳腺癌辅助诊断中的价值.方法 应用表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术检测113例乳腺癌患者、103例乳腺良性肿瘤患者及92例健康女性的血清蛋白质谱,采用Biomarker Pattern(BPS)软件分析蛋白质谱,建立分类树模型,然后对模型进行盲筛验证.结果 比较乳腺癌患者与健康女性的血清蛋白质谱,筛选出12个差异蛋白质峰.经验证,所建立的分类树模型Ⅰ诊断乳腺癌的灵敏度为91.9%,特异度为81.2%.比较乳腺良性肿瘤患者与健康女性的血清蛋白质谱,筛选出11个差异蛋白质峰.经验证,所建立的分类树模型Ⅱ诊断乳腺良性肿瘤的灵敏度为87.9%,特异度为81.2%.比较乳腺癌与乳腺良性肿瘤患者的血清蛋白质谱,筛选出2个差异蛋白质峰.经验证,所建立的分类树模型Ⅲ诊断乳腺癌的灵敏度为81.8%,特异度为78.3%.应用这些差异蛋白及分类树模型,分别对93例CA15-3阴性乳腺癌患者与36例乳腺良性疾病患者的血清、20例CA15-3阳性乳腺癌患者与36例乳腺良性疾病患者的血清进行盲筛,诊断乳腺癌的敏感度和特异度分别为80.6%和91.7%、75.0%和91.7%,明显高于传统的乳腺癌标志物CA15-3,而CA15-3阴性与CA15-3阳性乳腺癌未见明显的差异蛋白质峰.结论 应用SELDI-TOF-MS技术可筛选出乳腺癌、乳腺良性肿瘤和健康女性血清蛋白质谱存在的差异蛋白质峰,据此建立的诊断模型可用于乳腺癌的辅助诊断.     

The proteomic analysis of cisplatin resistance in breast cancer cells

Resistance to cisplatin represents a major obstacle in the effective management of many cancers, including metastatic breast cancer. We aimed to gain further understanding of the mechanisms underlying development of cisplatin resistance using an in vitro cell line model. The MCF-7 breast cancer cell line and a novel derivative displaying significant resistance to cisplatin were analyzed using two-dimensional gel electrophoresis. The protein profiles were compared and 15 differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The downregulation of beta-tubulin type 3, cytokeratin 17, tropomyosin 1-alpha, peroxiredoxin 4, heat shock 27-kDa protein 1, glutathione-S-transferase mu 3, ribosomal protein P0, isocitrate dehydrogenase 3, and peptidyl-prolyl isomerase A isoform 1 was associated with cisplatin-resistant cells. In contrast, the expression of hydroxyprostaglandin dehydrogenase 15-(NAD), matrix metalloproteinase 9, heterogeneous nuclear ribonucleoprotein A3, proteasome beta 1 subunit, electron transfer flavoprotein beta-polypeptide isoform 1, and peptidyl-propyl isomerase B precursor was upregulated in cisplatin-resistant cells. The downregulation (at least twofold) of glutathione-S-transferase mu 3, cytokeratin 17, and peroxiredoxin 4 was confirmed by Western blotting. We have identified alterations in the expression levels of several proteins that may be associated with cisplatin resistance and are candidates for further validation in clinical samples.     

Interference of the IGF system as a strategy to inhibit breast cancer growth

Summary Experimental evidence suggests that human breast cancer cells can be regulated by the IGF-I and IGF-II present in the tumor stromal elements and/or by the endogenous tumor cell IGF-II in a paracrine or autocrine fashion. Thus, blockade of the receptor signalling pathway could lead to diminished tumor growth. Blockade of the type I IGF receptor by a monoclonal antibody (IR3) has been used as a strategy to demonstrate the importance of the IGF pathway. Although IR3 could not block serum-free growth of breast cancer cell lines, it could inhibit anchorage independent growth in most cell lines in the presence of serum. In vivo, IR3 administered at the time of tumor cell inoculation could inhibit MDA-MB-231 tumor formation in athymic mice; however, inhibition of established tumors was not seen. Moreover, IR3 could not inhibit tumor formation of the MCF-7 cell line in vivo. These results suggest that blockade of the type I IGF receptor can inhibit the growth of some breast cancer cells both in vitro and in vivo. Future anti-growth factor strategies include the combination of anti-IGF receptor antibodies with IGF neutralizing modalities, the dual blockade of growth factor receptors (epidermal growth factor receptor and type I IGF receptor), and combinations of steroid hormone antagonists and anti-growth factor treatments to maximize tumor inhibition.     

Age as a prognostic factor in breast cancer: Relationship to pathologic and biologic features

The relationship of age with prognostic factors and outcome of breast cancer has long been controversial due to numerous confounding factors. In order to clarify the prognostic value of age, we analyzed a homogeneous population of 1,266 patients treated for breast cancer at the same institution (mean follow-up: 62 months). Three groups were compared: patients under 35 years of age, non-menopausal patients over 35 years of age, and post-menopausal patients under the age of 70 years. A higher frequency of undifferentiated tumors, histoprognostic grade-3 cancer, microscopic lymph-node involvement and negative hormonal receptor status was observed in patients under 35 years. In addition, clinical but not anatomical tumor size was greater in young patients, suggesting higher stromal activity. Metastasis-free survival and overall survival were significantly poorer before 35 years. Differences were observed when patients were matched with regard to stage, anatomic size, histoprognostic grade, microscopic lymph-node involvement and receptor status. Multivariate analysis of both overall and metastasis-free survival demonstrated that age younger than 35 years was an independent risk factor. Younger women had a higher risk of local recurrence but, unlike older women, they did not experience any worsening of the already unfavorable outcome due to recurrence. © 7995 Wiley-Liss, Inc.     

乳腺癌临床特征与预后分析

目的：总结经综合治疗的233例乳腺癌患者的临床病理特征及生存情况,探讨影响乳腺癌预后的因素。方法：建立乳腺癌患者的临床资料库,采用SPSS8.0统计软件对临床数据进行单因素及多因素生存分析,以发现影响乳腺癌长期生存的因素。结果：所有患者1、2、3、5和8年生存率分别为95.71%（223/233）、82.83%（193/233）、61.37%（143/233）、37.34%（87/233）和6.87%（16/233）。单因素分析显示腋淋巴结转移数目;原发肿瘤大小与生存呈负相关（P均〈0.001）;TNM晚期患者复发转移的患者中位生存明显缩短（P均〈0.0001）;术后辅助化疗方案选择含有蒽环类的方案组生存期优于不含有蒽环类的方案组（P〈0.05,X^2=9.99）;病理类型治疗方式与生存时间相关（P均〈0.01）。COX比例风险模型分析显示治疗方式、术后辅助性化疗方案和复发转移具有独立预后意义（P〈0.05）。Logistic分析结果显示,原发肿瘤大小、腋窝淋巴结转移状态、病理类型、综合治疗方式是影响乳腺癌复发转移的主要因素,其影响程度的排序依次是腋窝淋巴结转移、原发肿瘤大小、病理类型、综合治疗方式（P〈0.0001）。结论：乳腺癌患者的远期生存及复发转移与原发肿瘤大小、腋窝淋巴结转移状态、病理类型、术后化疗方案的选择及个体化综合治疗方式密切相关。     

Identification of 14-3-3sigma as a contributor to drug resistance in human breast cancer cells using functional proteomic analysis

Multidrug resistance (MDR) is a major obstacle to successful cancer treatment. To understand the mechanism of MDR, many cancer cell lines have been established, and various mechanisms of resistance, such as ATP-binding cassette (ABC) transporter-mediated drug efflux, have been discovered. Previously, a MDR cell line MCF7/AdVp3000 was selected from breast cancer cell line MCF7 against Adriamycin, and overexpression of ABCG2 was thought to cause MDR in this derivative cell line. However, ectopic overexpression of ABCG2 in MCF7 cells could not explain the extremely high drug resistance level of the selected MCF7/AdVp3000 cells. We hypothesized that MCF7/AdVp3000 cells must have other resistance mechanisms selected by Adriamycin. To test this hypothesis, we compared the global protein profiles between MCF7 and MCF7/AdVp3000 cells. Following two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry analysis, 17 protein spots with differential levels between the two cell lines were identified. Although 14-3-3sigma, keratin 18, keratin 19, ATP synthase beta, protein disulfide isomerase, heat shock protein 27, cathepsin D, triose-phosphate isomerase, peroxiredoxin 6, and electron transfer flavoprotein were increased, nm23/H1, peroxiredoxin 2, nucleophosmin 1/B23, and inorganic pyrophosphatase were decreased in MCF7/AdVp3000 cells. The differential levels of these proteins were validated using Western blot. Furthermore, functional validation showed that the elevated 14-3-3sigma expression contributes considerably to the observed drug resistance in MCF7/AdVp3000 cells. We, thus, conclude that these proteins likely contribute to the resistance selected in the MCF7/AdVp3000 cells, and their altered expression in tumors may cause clinical resistance to chemotherapy.     

Systematic high-content proteomic analysis reveals substantial immunologic changes in colorectal cancer

The immune system is a significant determinant of epithelial tumorigenesis, but its role in colorectal cancer pathogenesis is not well understood. The function of the immune system depends upon the integrity of the protein network environment, and thus, we performed MELC immunofluorescence microscopy focusing on the lamina propria. By analyzing structurally intact tissues from colorectal cancer, ulcerative colitis, and healthy colonic mucosa, we used this unique and novel highly multiplexed robotic-imaging technology, which allows visualizing dozens of proteins simultaneously, and explored the toponome in colorectal cancer mucosa for the first time. We identified 1,930 motifs that distinguish control from colorectal cancer tissue. In colorectal cancer, the number of activated T cells is increased, explained by a lack of bax, caspase-3, and caspase-8. Whereas CD4(+)CD25(+) T cells are decreased and are, other than in ulcerative colitis, not activated, cytotoxic T cells are significantly increased in colorectal cancer. Furthermore, the number of activated human lymphocyte antigen (HLA)-DR(+) T-cells is increased in colorectal cancer, pointing to an altered antigen presentation. In colorectal cancer, CD3(+)CD29(+) expression and assembly of the LFA-1 and LFA-3 receptor are differentially changed, indicating a distinct regulation of T-cell adhesion in colorectal cancer. We also identified increased numbers of natural killer and CD44(+) cells in the colorectal cancer mucosa and nuclear factor-kappaB as regulator of apoptosis in these cell populations. High-content proteomic analysis showed that colorectal cancer induces a tremendous modification of protein expression profiles in the lamina propria. Thus, topological proteomic analysis may help to unravel the role of the adaptive immune system in colorectal cancer and aid the development of new antitumor immunotherapy approaches.     

Using a population-based cancer reporting system to evaluate a breast cancer detection and awareness program.

During May 1987, a total of 10,207 Wisconsin women were screened as part of a statewide Breast Cancer Detection and Awareness program sponsored by the Wisconsin Division of the American Cancer Society. Data from a population-based cancer reporting system were used to predict the number of expected breast cancer cases for that year. After controlling for secular and seasonal trends, we found that, compared with the number of cases that would have been expected, 51 more cases of localized breast cancer were diagnosed in the state during the time of the program. This study demonstrates the public health impact of a statewide screening program and the usefulness of a cancer reporting system in program evaluation.     

Inflammation and breast cancer. Metalloproteinases as common effectors of inflammation and extracellular matrix breakdown in breast cancer

Two rapidly evolving fields are converging to impact breast cancer: one has identified novel substrates of metalloproteinases that alter immune cell function, and the other has revealed a role for inflammation in human cancers. Evidence now shows that the mechanisms underlying these two fields interact in the context of breast cancer, providing new opportunities to understand this disease and uncover novel therapeutic strategies. The metalloproteinase class of enzymes is well studied in mammary gland development and physiology, but mostly in the context of extracellular matrix modification. Aberrant metalloproteinase expression has also been implicated in breast cancer progression, where these genes act as tumor modifiers. Here, we review how the metalloproteinase axis impacts mammary physiology and tumorigenesis and is associated with inflammatory cell influx in human breast cancer, and evaluate its potential as a regulator of inflammation in the mammary gland.     

Identification of ganglioside GM2 activator playing a role in cancer cell migration through proteomic analysis of breast cancer secretomes

Cancer cell secretomes are considered a potential source for the discovery of cancer markers. In this study, the secretomes of four breast cancer (BC) cell lines (Hs578T, MCF‐7, MDA‐MB‐231, and SK‐BR‐3) were profiled with liquid chromatography–tandem mass spectrometry analysis. A total of 1410 proteins were identified with less than 1% false discovery rate, of which approximately 55% (796 proteins) were predicted to be secreted from cells. To find BC‐specific proteins among the secreted proteins, data of immunohistochemical staining compiled in the Human Protein Atlas were investigated by comparing the data of BC tissues with those of normal tissues. By applying various criteria, including higher expression level in BC tissues, higher predicted potential of secretion, and sufficient number of tandem mass spectra, 12 biomarker candidate proteins including ganglioside GM2 activator (GM2A) were selected for confirmation. Western blot analysis and ELISA for plasma samples of healthy controls and BC patients revealed elevation of GM2A in BC patients, especially those who were estrogen receptor‐negative. Additionally, siRNA‐mediated knockdown of GM2A in BC cells decreased migration in vitro, whereas the overexpression of GM2A led to an increase in cell migration. Although GM2A as a diagnostic and prognostic marker in BC should be carefully verified further, this study has established the potential role of GM2A in BC progression.     

基因分型在乳腺癌个体化治疗中的应用

临床上经常可以观察到组织学类型、临床分期,甚至激素受体状态都相同的乳腺癌患者,采用同样的治疗方案,治疗反应和预后却常有很大差异。这说明乳腺癌是一类高度异质性的肿瘤。目前学者们认为,这种现象是由于肿瘤的分子差异所致,但在临床上一直没有很好的对策。肿瘤分子病理学的发展以及由此产生的乳腺癌基因筛选技术,为解决这一问题提供了希望。本文综述乳腺癌相关基因及其表达产物检测对临床治疗的影响。     

乳腺癌不同分子亚型的临床特点和生存分析

目的探讨乳腺癌各分子亚型的临床特点及其预后情况。方法回顾性分析482例可手术乳腺癌患者资料．以免疫组织化学技术为基础,把乳腺癌分为4种分子亚型：luminalA型[ER（＋）或PR（＋）且HER-2（-）],luminalB型[ER（＋）或pR（＋）且HER-2（＋）],HER-2过表达型[ER（-）、PR（-）且HER-2（＋）j和basal—like型[ER（-）、PR（-）且HER-2（-）],并分析其临床特点及预后情况。结果全组共482例,其中luminalA型占46．1％（222／482）,luminalB型占14．7％（71／482）,HER-2过表达型占10．4％（50／482）,basal—like型占28．8％（139／482）。运用x。检验各分子亚型在年龄、月经状况、肿瘤大小、淋巴结状况和临床分期等的差异均无统计学意义。全组有完整随访资料者共441例,中位随访时间62个月。随访结果显示．HER-2过表达型和basal—like型的远处转移率均高于luminalA型,且差异有统计学意义（x2=11．659,P=0．009）;运用Kaplan—Meier法分析各分子亚型的生存预后,luminalA型的无病生存率、无远处转移生存率和总生存率最高,HER-2过表达型和basal-like型的预后最差,差异有统计学意义（Log—Rank检验,P均〈0．050）。结论乳腺癌分子分型对患者预后的判断具有重要临床意义,有望成为今后制定乳腺癌个体化治疗的重要依据。     

Nuclear polymorphism — a prognostic parameter to evaluate local recurrence of female breast cancer

Summary Between 1980 and 1986 676 patients underwent surgery for primary breast cancer. Of these, 35 patients developed locoregional recurrence. Retrospective analysis of the spontaneous postoperative development revealed 2 groups: group A had subsequent distant metastases, group B was tumor-free after surgical treatment of local recurrence. Analysis of the commonly employed characterization criteria of primary tumors (tumor size, lymph node involvement, estrogen receptors, histologic grading of primary tumors, and excised locoregional recurrence) showed no statistically significant difference between the two groups. However, a more detailed differentiation of the subcriteria for the histologic grading according to Bloom and Richardson revealed a prevalence of anaplastic nuclei in the primary tumors of group A (9/11). Even in this small patient population the parameter of nuclear polymorphism revealed a highly significant statistical difference between the two groups.     

男性乳腺癌25例的临床病理特征及生存分析

目的分析男性乳腺癌患者的临床病理特征、治疗及生存情况。方法回顾性分析北京大学肿瘤医院乳腺肿瘤内科及北京大学第鼍医院收治的25例男性乳腺癌的临床及随访资料。采用Kaplan．Meier模型分析男性乳腺癌生存情况。结果分析的25例男性乳腺癌发病中位年龄为65岁（44—80岁）。病理类型多为浸润性导管癌（80．0％）,仅l例患者为浸润性小叶癌（4．0％）。激素受体阳性患者23例（92．O％）。luminalA—like亚型12例（48．0％）,Her-2阴性luminalB—like亚型2例（8．0％）,Her-2阳性luminal B—like亚型6例（24．0％）,无HER-2过表达型,basal—like型（i阴型）2例（8．0％）,未分型3例（12．O％）。早期（I～Ⅲ期）患者比例较大,占92％（23例）,均进行了根治性手术治疗,9例（39．1％）接受了辅助化疗,14例（60．9％）接受了辅助内分泌治疗,其中7例患者为他莫昔芬治疗,4例患者为芳香化酶抑制剂（aronlatase inhibitors,AI）联合戈舍瑞林治疗,3例患者为他莫昔芬治疗2～3年转换为AI联合戈舍瑞林治疗。早期乳腺癌患者的5年无病生存率为81％。结论男性乳腺癌是一种少见的疾病,以老年患者为主。早期患者治疗以手术为主,内分泌治疗原则不能完全相同于绝经后的女性乳腺癌,使用AI治疗的患者,应注意下丘脑-垂体-睾丸轴的功能抑制。