Prognostic value of Ki-67 proliferating index in triple negative breast carcinomas

The aim of this study was to assess Ki-67 in the triple negative breast cancer group (TNBC) in addition to basal like (BL) immunophenotype, BL morphology and conventional clinicopathologic factors, and to demonstrate their prognostic relevance in this group of tumors.     

ki67在三阴性乳腺癌中的临床病理意义

目的 通过检测三阴性乳腺癌中细胞增殖抗原ki67的表达情况,分析其与临床病理特征及预后的关系,明确ki67是否可以作为判断三阴性乳腺癌的预后指标.方法 应用免疫组织化学方法检测ki67在三阴性乳腺癌组织中的表达,分析ki67表达与三阴性乳腺癌患者临床病理特征的相关性及与预后的关系.结果 ki67在三阴性乳腺癌组织中的表达与肿瘤大小、TNM分期、组织学分级和淋巴结转移有关(x2=10.536、16.824、11.020、7.180、P＜0.05).单因素生存分析结果显示:肿瘤大小、临床分期、组织学分级、淋巴结转移和ki67与患者总生存均相关(OR=4.211、3.800、0.288、3.502、2.612,P＜0.05).Cox多因素生存分析显示,淋巴结转移与总生存有一定的相关性(OR=2.768,P＜0.05).结论 ki67表达与三阴性乳腺癌中肿瘤大小、TNM分期、组织学分级和淋巴结转移有关,对于三阴性乳腺癌的预后评估具有一定的价值,可作为评价三阴性乳腺癌预后的候选指标.通过Cox多因素生存分析,淋巴结转移可作为与总生存相关的独立预后因素.     

三阴性乳腺癌干细胞微球体的治疗

三阴性乳腺癌(TNBC)是指雌激素受体(ER)、孕激素受体(PR)及人类表皮生长因子受体2 (HER-2)均为阴性的乳腺癌。TNBC恶性程度高,
侵袭性强,易远处转移,预后差,对内分泌治疗及靶向治疗的效果均欠佳,是临床治疗的难点。近几年研究发现,三阴性乳腺癌的发生、发展
很可能与其中的乳腺癌干细胞有关,乳腺癌干细胞微球体培养是一种收集干细胞的方法,被广泛用于三阴性乳腺癌的干细胞研究。     

GABRP基因在基底样三阴乳腺癌中的差异表达

目的:探索GABRP基因在基底样三阴乳腺癌(basal-like Triple negative breast cancer,BLTNBC)中的表达,为临床提供预后诊断分子标志物.方法:通过生物信息学分析比较GABRP基因的表达;采用定量PCR方法检测GABRP基因在乳腺癌细胞系及104例乳腺癌石蜡样本中的差异表达,并采用相关性分析、卡方检验及Fisher精确概率法分析GABRP基因与细胞增殖、淋巴结转移、ER及HER-2基因表达的相关性.结果:生物信息学分析和定量PCR结果显示GABRP基因在三阴性乳腺癌(triple negative breast cancer,TNBC)及BLTNBC中显著性高表达(p＜0.05);在non-TNBC,TNBC,BLTNBC中,其表达与细胞增殖、HER-2表达无明显相关性(P＞0.05),而在BLTNBC中与淋巴结转移状态有明显相关性(P＜0.05),在non-TNBC巾与ER表达有明显相关性(P＜0.05).结论:GABRP基因可作为BLTNBC的有潜力的预后诊断分子标志物.     

c-Met对三阴性乳腺癌细胞增殖及阿霉素耐药性的影响

目的:研究c-Met对三阴性乳腺癌细胞株MDA-MB-231活力及对阿霉素耐药性的影响。方法:构建阿霉素耐药的MDA-MB-231/ADR细胞系,实时荧光定量PCR及Western blotting技术检测不同细胞系中c-Met mRNA及蛋白的表达。脂质体转染c-Met-siRNA及表达质粒或AKT-siRNA,Western blotting检测转染效率;四甲基偶氮唑法(MTT法)检测细胞的活力及对阿霉素的敏感性。结果:对阿霉素耐药的MDA-MB-231/ADR细胞中cMet的mRNA及蛋白表达均显著高于对照的MDA-MB-231细胞,转染高表达c-Met的质粒可增加MDA-MB-231细胞的活力并降低其对阿霉素的敏感性,而利用siRNA抑制耐药细胞株中c-Met的表达后,可以逆转MDA-MB-231/ADR细胞对阿霉素的耐药。此外,c-Met可以磷酸化激活细胞中的AKT,并通过该信号分子增加MDA-MB-231细胞活力并诱导耐药。结论:c-Met可作为一个重要的靶点应用于三阴性乳腺癌的治疗。     

An associated classification of triple negative breast cancer: the risk of relapse and the response to chemotherapy

Background: Triple negative breast cancer (TNBC) is heterogeneous and considered as an aggressive tumor. This study was to evaluate the associated classification and its correlations with prognosis and the response to chemotherapy in Chinese women. Methods: Four hundred and twenty-eight cases of invasive TNBC were involved in this study. The expression of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), epidermal growth factor receptor (EGFR), and cytokeratin 5/6 (CK5/6), Ki67 and p53 were analyzed by immunohistochemistry and compared with patient outcome, and its implications and chemotherapy response were evaluated in four subgroups: typical medullary carcinoma (TMC), atypical medullary carcinoma (AMC), non-specific invasive ductal carcinoma (IDC) and other types. Results: The factors of tumor grade, tumor stage, lymph node status, EGFR/CK5/6 status and p53 labeling index were different among the groups. TMC tumors had the lowest rate of relapse (5.8%), while AMC, IDC and other types were associated with an increased risk of relapse (19.1%, 26.7% and 38.2% respectively). Many factors were risk predictors of relapse for TNBC and IDC, while only positive lymph node was for AMC. For MC tumors, adjunctive chemotherapy decreased the risk of relapse in lymph node positive subgroup (36.8% and 66.7%), while not significant in lymph node negative one (8.1% and 10.0%). Conclusion: The classification based on histologic and IHC findings may be a significant improvement in predicting outcome in TNBC. The different chemotherapy response in subgroups may contribute to guiding the treatment of TNBC.     

C-kit and PDGFRA gene mutations in triple negative breast cancer

In this study, we evaluated C-kit immunohistochemical expression and C-kit and platelet derived growth factor receptor A (PDGFRA) gene mutations in triple negative breast cancer. 171 cases were analyzed by immunohistochemical staining for the expression of C-kit and 45 cases, including 10 C-kit negative cases and 35 C-kit positive cases, were performed for C-kit gene mutations in exons 9, 11, 13 and 17 and PDGFRA gene mutations in exons 12 and 18. C-kit expression was detected in 42.1% of triple negative breast cancers. Only 1 activating mutation was detected in exon 11 of C-kit gene in 1 case. No activating mutations were found in the other 44 cases. C-kit expression is a frequent finding in triple negative breast cancers; 1 activating mutation which was also found in gastrointestinal stromal tumors was detected; a few cases might benefit from imatinib.     

三阴型乳腺癌中BRCA1的表达及其预后意义

目的：探讨乳腺癌易感基因1(breast cancer susceptibility gene 1, BRCA1)蛋白的表达及其对三阴型乳腺癌(triple nega-tive breast cancer, TNBC)患者预后的影响。方法采用免疫组化SP法检测95例TNBC组织蜡块中BRCA1、p53的表达,并结合患者临床病理特征进行相关性分析和预后评价。结果 TNBC中BRCA1的阳性率为31．6%,阳性患者中位年龄较阴性者明显偏小(P=0．047),且p53表达明显升高(P=0．001),阳性者与阴性者的总生存期差异无显著性(HR=1．110,95%CI=0．552~2．235,P=0．769)。结论 BRCA1蛋白表达与TNBC患者发病年龄、p53表达有关,但对患者的预后无明显影响。 p53通路与BRCA1表达在抑制肿瘤生长方面可能存在相关性。     

Phosphorylated mTOR expression correlates with poor outcome in early-stage triple negative breast carcinomas

The mammalian target of rapamycin (mTOR) plays an important role in cell growth, proliferation, and metabolism. Some studies have associated phosphorylated mTOR (p-mTOR) expression with worse outcome in breast cancers. However, the significance of p-mTOR expression specifically in triple negative breast carcinoma (TNBC) is unknown. In this study, p-mTOR expression was evaluated by immunohistochemistry in 172 TNBCs and the result was correlated with clinicopathologic variables and disease outcome. The majority of tumors (72.1%) were p-mTOR positive; p-mTOR expression did not correlate with age, tumor size, grade, lymph node status, or tumor stage. In patients at stage 1 and 2 disease, those with p-mTOR expression had significantly worse overall as well as recurrence-free survival compared to those without p-mTOR expression. p-mTOR expression appears to be an adverse prognostic indicator in early-stage TNBCs. The assessment of p-mTOR expression in these tumors may also help to stratify patients for future target therapy studies.     

RNA-seq analysis identified hormone-related genes associated with prognosis of triple negative breast cancer

Triple negative breast cancer (TNBC) is an aggressive subtype of breast cancer that currently lacks effective biomarkers and therapeutic targets required to investigate the diagnosis and treatment of TNBC. Here we performed a comprehensive differential analysis of 165 TNBC samples by integrating RNA-seq data of breast tumor tissues and adjacent normal tissues from both our cohort and The Cancer Genome Atlas (TCGA). Pathway enrichment analysis was conducted to evaluate the biological function of TNBC-specific expressed genes. Further multivariate Cox proportional hazard regression was performed to evaluate the effect of these genes on TNBC prognosis. In this report, we identified a total of 148 TNBC-specific expressed genes that were primarily enriched in mammary gland morphogenesis and hormone levels related pathways, suggesting that mammary gland morphogenesis might play a unique role in TNBC patients differing from other breast cancer types. Further survival analysis revealed that nine genes (FSIP1, ADCY5, FSD1, HMSD, CMTM5, AFF3, CYP2A7, ATP1A2, and C11orf86) were significantly associated with the prognosis of TNBC patients, while three of them (ADCY5, CYP2A7, and ATP1A2) were involved in the hormone-related pathways. These findings indicated the vital role of the hormone-related genes in TNBC tumorigenesis and may provide some independent prognostic markers as well as novel therapeutic targets for TNBC.     

三阴型乳腺癌中RHBDD1和EGFR的表达及相关性分析

目的探讨RHBDD1和EGFR在三阴型乳腺癌(triple negative breast cancer, TNBC)发生、转移中的作用及其临床意义。方法选取30例TNBC(ER、PR、HER2均阴性)和30例非三阴型乳腺癌(non triple negative breast cancer, NTNBC)(ER、PR、HER2至少一项阳性)患者作为研究对象,采用免疫组化、酶联免疫法和qRT-PCR技术检测组织和血清中RHBDD1和EGFR mRNA含量及蛋白表达。结果 TNBC组织中RHBDD1和EGFR蛋白阳性率(83%、80%)高于NTNBC组织(67%、37%),差异均有统计学意义(P<0.05);TNBC患者血清中RHBDD1和EGFR的浓度含量亦明显高于NTNBC患者,差异均有统计学意义(P<0.05);TNBC组织中的RHBDD1 mRNA和EGFR mRNA含量分别是癌旁组织的3.68倍和4.56倍,差异均有统计学意义(P<0.05)。RHBDD1和EGFR的表达与TNBC患者年龄无关,与临床分期、组织学分级、肿瘤大小、淋巴结转移以及远处转移呈正相关。...     

三阴性乳腺癌整合素连接激酶表达的预后意义

目的：探讨三阴性乳腺癌中整合素连接激酶（integrin-linked-kinase,ILK）的表达与患者预后的关系。方法：应用免疫组织化学SP法检测ILK在60例三阴性乳腺癌、36例非三阴性乳腺癌中的表达情况,并结合随访资料评价ILK表达水平对预测三阴性乳腺癌患者预后的价值。结果：三阴性乳腺癌中ILK的表达显著高于非三阴性乳腺癌（P〈0.01）;其ILK阳性表达与肿瘤大小、腋窝淋巴结转移、临床分期有关,而与患者年龄、月经状态无关;Kaplan-Meier生存曲线显示,ILK高表达组复发率高,病死率高,预后差。结论：ILK表达水平可作为判断三阴性乳腺癌患者预后的指标之一。     

XAV-939对三阴性乳腺癌抑制的作用

目的:探索XAV-939对三阴性乳腺癌的生长产生抑制作用及其机制.方法:体外检测XAV-939对MDA-MB-231细胞的增殖抑制作用,并检测Axin 1、Axin 2、β-catenin和p-β-catenin蛋白的表达以及VEGF-A的分泌水平.在体观察XAV-939对裸鼠移植瘤的体积和瘤重变化情况,并检测肿瘤组织中Axin 1、Axin 2、β-catenin和p-β-catenin蛋白的表达,以及VEGF表达、微血管密度、增殖指数及凋亡指数.结果:XAV-939可以抑制MDA-MB-231细胞的生长,其抑制作用呈浓度依赖型;同时XAV-939抑制了MDA-MB-231细胞分泌VEGF-A的能力.体内实验中,XAV-939可以明显抑制肿瘤组织生长;肿瘤组织的VEGF表达量下降,微血管密度下降;肿瘤组织的凋亡指数显著上升;增殖指数没有明显变化.体外体内实验均显示XAV-939处理组的Axin 1、Axin 2和p-β-catenin表达量明显上升而β-catenin显著下降.结论:XAV-939可以通过抑制Wnt/β-catenin通路在体内外抑制三阴性乳腺癌细胞/组织的增殖.     

Expression of MHC class I and class II antigens in primary breast carcinomas and synchronous nodal metastases

Expression of the major histocompatibility (MHC) class I and class II antigens was studied by immunohistochemistry in a series of 70 primary breast carcinomas and in nodal metastases. In particular, the expression of class I (HLA A-B-C) and class II (DP, DQ and DR) molecules was compared in: a) primary breast cancers devoid of nodal metastases (n = 36) and tumors exhibiting metastatic deposits (n = 34) at the time of surgery, and b) primary breast carcinomas and their corresponding synchronous axillary nodal metastases. Reduced or absent HLA A-B-C antigen expression was seen in approximately 54.3% of primary breast carcinomas, whereas a partial or complete induction of class II products was observed in 18.5% (DQ), 30% (DP) or 48.5% (DR) of the same cases. An almost complete overlap of antigen expression was observed in breast tumors in which no metastases were found by histological examination of axillary nodes and in neoplasms showing histologically-diagnosed synchronous metastases. The reactivity for class I and class II antigens in nodal metastases roughly paralleled that exhibited by corresponding primary tumors. A discordant expression was seen in 11 cases (32%) stained for HLA A-B-C and in 8 (24%), 7 (21%) and 6 (18%) cases assayed for DP, DQ and DR products, respectively. When a discordant expression was detected, either decreased or increased staining patterns were observed in metastases. The finding of overlapping MHC antigenic profiles in the majority of primary breast tumors and nodal metastases casts doubts on the hypothesis that loss of MHC antigens can play an important role in the seeding and growth of metastatic breast carcinoma cells.     

Nucleolin overexpression in breast cancer cell sub-populations with different stem-like phenotype enables targeted intracellular delivery of synergistic drug combination

Breast cancer stem cells (CSC) are thought responsible for tumor growth and relapse, metastization and active evasion to standard chemotherapy. The recognition that CSC may originate from non-stem cancer cells (non-SCC) through plastic epithelial-to-mesenchymal transition turned these into relevant cell targets. Of crucial importance for successful therapeutic intervention is the identification of surface receptors overexpressed in both CSC and non-SCC. Cell surface nucleolin has been described as overexpressed in cancer cells as well as a tumor angiogenic marker. Herein we have addressed the questions on whether nucleolin was a common receptor among breast CSC and non-SCC and whether it could be exploited for targeting purposes.Liposomes functionalized with the nucleolin-binding F3 peptide, targeted simultaneously, nucleolin-overexpressing putative breast CSC and non-SCC, which was paralleled by OCT4 and NANOG mRNA levels in cells from triple negative breast cancer (TNBC) origin. In murine embryonic stem cells, both nucleolin mRNA levels and F3 peptide-targeted liposomes cellular association were dependent on the stemness status. An in vivo tumorigenic assay suggested that surface nucleolin overexpression per se, could be associated with the identification of highly tumorigenic TNBC cells. This proposed link between nucleolin expression and the stem-like phenotype in TNBC, enabled 100% cell death mediated by F3 peptide-targeted synergistic drug combination, suggesting the potential to abrogate the plasticity and adaptability associated with CSC and non-SCC.Ultimately, nucleolin-specific therapeutic tools capable of simultaneous debulk multiple cellular compartments of the tumor microenvironment may pave the way towards a specific treatment for TNBC patient care.     

Spontaneous CD4+ and CD8+ T‐cell responses directed against cancer testis antigens are present in the peripheral blood of testicular cancer patients

Cancer/testis antigen (CTAg) expression is restricted to spermatogenic cells in an immune‐privileged site within the testis. However, these proteins are expressed aberrantly by malignant cells and T‐cell responses against CTAgs develop in many cancer patients. We investigated the prevalence, magnitude and phenotype of CTAg‐specific T cells in the blood of patients with testicular germ cell tumors (TGCTs). CD8⁺ and CD4⁺ T‐cell responses against MAGE‐A family antigens were present in 44% (20/45) of patients’ samples assayed by ex vivo IFN‐γ ELISPOT. The presence of MAGE‐specific CD8⁺ T cells was further determined following short‐term in vitro expansion through the use of pMHC‐I multimers containing known immunogenic peptides. Longitudinal analysis revealed that the frequency of MAGE‐specific T cells decreased by 89% following orchidectomy suggesting that persistence of tumor antigen is required to sustain CTAg‐specific T‐cell immunity. Notably, this decrease correlated with a decline in the global effector/memory T‐cell pool following treatment. Spontaneous T‐cell immunity against CTAg proteins therefore develops in many patients with testicular cancer and may play an important role in the excellent clinical outcome of patients with this tumor subtype.     

Increased tumor cell expression of Axl is a marker of aggressive features in breast cancer among African women

Axl, a receptor tyrosine kinase belonging to the Tyro/Axl/Mer (TAM) family, has been shown to be overexpressed in breast cancer with poor outcome. Moreover, Axl was associated with a basal‐like phenotype (BLP) in these tumors. Our aim was to investigate Axl expression in breast cancers from an African population since these tumors are known to be aggressive and have a high frequency of the basal‐like phenotype. We studied 170 paraffin‐embedded breast carcinoma cases by tissue microarrays and immunohistochemical methods. In total, 128 tumor cases (75%) had strong Axl expression and 42 cases (25%) had weak or negative staining. Strong expression of Axl was associated with high tumor grade (p < 0.0005), estrogen receptor (ER) negativity (p = 0.024), p53 expression (p = 0.004), P‐cadherin positivity (p = 0.017), and basal‐like phenotypic profiles BLP2 (p = 0.033) and BLP3 (p = 0.022). In addition, Axl overexpression also showed an association with markers of tumor cell proliferation and tumor angiogenesis. In conclusion, our findings indicate strong expression of Axl in a high proportion of breast cancer cases among African women and associations with markers of aggressive features, indicating poor prognosis. These findings suggest Axl as a potential therapeutic target in this population.