核酸适配体在三阴性乳腺癌诊疗中的研究进展*

乳腺癌是女性高发恶性肿瘤,三阴性乳腺癌(triple-negative breast cancer, TNBC)恶性程度极高,且发病机制复杂,是乳腺癌分型中预后最差的类型,但目前其早期筛查和诊断的敏感度仍处在较低水平。因此,亟须通过应用具有高度特异性的肿瘤标志物分子探针,实现其早期诊断和治疗。核酸适配体是在人工合成的随机单链核酸序列文库中,通过指数富集的配体系统进化技术(systematic evolution of ligands by exponential enrichment, SELEX)筛选获得的寡核苷酸序列。高效的分子识别能力使其成为最具潜力的生物靶向分子,在肿瘤诊断及治疗中具有广阔的应用前景。目前,通过筛选已获得了多种靶向TNBC细胞的核酸适配体。重点综述基于SELEX及其衍生技术筛选TNBC相关核酸适配体的新进展,以及核酸适配体在TNBC诊断和治疗中的应用,为相关研究提供参考。     

三阴型乳腺癌的化学治疗和靶向治疗进展

三阴型乳腺癌(Triple negative Breast cancer,TNBC)占乳腺癌总数的15%,是一种免疫组织化学亚型。通常发生于青年女性,有很高的复发率,内脏和中枢神经系统转移早,病程短、死亡率高。对大多数TNBC患者,常规化疗是主要的治疗方式。20%患者有很好的化疗敏感性,对化疗耐药的TNBC患者靶向治疗为当前研究的热点,寻找乳腺癌新的治疗靶点,提出TNBC新的治疗策略,有望去改善TNBC患者的预后。     

Elevated Protein Kinase D3 (PKD3) Expression Supports Proliferation of Triple-negative Breast Cancer Cells and Contributes to mTORC1-S6K1 Pathway Activation

Here, we show that the expression of the Golgi-localized serine-threonine kinase protein kinase D3 (PKD3) is elevated in triple-negative breast cancer (TNBC). Using an antibody array, we identified PKD3 to trigger the activation of S6 kinase 1 (S6K1), a main downstream target of the mammalian target of rapamycin complex 1 (mTORC1) signaling pathway. Accordingly, PKD3 knockdown in TNBC cells led to reduced S6K1 phosphorylation, which was associated with impaired activation of mTORC1 at endolysosomal membranes, the accumulation of the mannose 6-phosphate receptor in and the recruitment of the autophagy marker light chain 3 to enlarged acidic vesicles. We further show that PKD3 depletion strongly inhibited cell spreading and proliferation of TNBC cells, identifying this kinase as a potential novel molecular therapeutic target in TNBC. Together, our data suggest that PKD3 in TNBC cells provides a molecular connection between the Golgi and endolysosomal compartments to enhance proliferative mTORC1-S6K1 signaling.     

Accuracy and completeness of the New Zealand Cancer Registry for staging of invasive breast cancer

PurposePopulation based cancer registries are an invaluable resource for monitoring incidence and mortality for many types of cancer. Research and healthcare decisions based on cancer registry data rely on the case completeness and accuracy of recorded data. This study was aimed at assessing completeness and accuracy of breast cancer staging data in the New Zealand Cancer Registry (NZCR) against a regional breast cancer register.MethodologyData from 2562 women diagnosed with invasive primary breast cancer between 1999 and 2011 included in the Waikato Breast Cancer Register (WBCR) were used to audit data held on the same individuals by the NZCR. WBCR data were treated as the benchmark.ResultsOf 2562 cancers, 315(12.3%) were unstaged in the NZCR. For cancers with a known stage in the NZCR, staging accuracy was 94.4%. Lower staging accuracies of 74% and 84% were noted for metastatic and locally invasive (involving skin or chest wall) cancers, respectively, compared with localized (97%) and lymph node positive (94%) cancers. Older age (>80 years), not undergoing therapeutic surgery and higher comorbidity score were significantly (p < 0.01) associated with unstaged cancer. The high proportion of unstaged cancer in the NZCR was noted to have led to an underestimation of the true incidence of metastatic breast cancer by 21%. Underestimation of metastatic cancer was greater for Māori (29.5%) than for NZ European (20.6%) women. Overall 5-year survival rate for unstaged cancer (NZCR) was 55.9%, which was worse than the 5-year survival rate for regional (77.3%), but better than metastatic (12.9%) disease.ConclusionsUnstaged cancer and accuracy of cancer staging in the NZCR are major sources of bias for the NZCR based research. Improving completeness and accuracy of staging data and increasing the rate of TNM cancer stage recording are identified as priorities for strengthening the usefulness of the NZCR.     

Lamin B2 promotes the progression of triple negative breast cancer via mediating cell proliferation and apoptosis

Triple negative breast cancer (TNBC) is a more common type of breast cancer with high distant metastasis and poor prognosis. The potential role of lamins in cancer progression has been widely revealed. However, the function of lamin B2 (LMNB2) in TNBC progression is still unclear. The present study aimed to investigate the role of LMNB2 in TNBC. The cancer genome atlas (TCGA) database analysis and immunohistochemistry (IHC) were performed to examine LMNB2 expression levels. LMNB2 short hairpin RNA plasmid or lentivirus was used to deplete the expression of LMNB2 in human TNBC cell lines including MDA-MB-468 and MDA-MB-231. Alterations in cell proliferation and apoptosis in vitro and the nude mouse tumorigenicity assay in vivo were subsequently analyzed. The human TNBC tissues shown high expression of LMNB2 according to the bioinformation analysis and IHC assays. LMNB2 expression was correlated with the clinical pathological features of TNBC patients, including pTNM stage and lymph node metastasis. Through in vitro and in vivo assays, we confirmed LMNB2 depletion suppressed the proliferation and induced the apoptosis of TNBC cells, and inhibited tumor growth of TNBC cells in mice, with the decrease in Ki67 expression or the increase in caspase-3 expression. In conclusion, LMNB2 may promote TNBC progression and could serve as a potential therapeutic target for TNBC treatment.     

彩色多普勒超声赋分诊断三阴性乳腺癌并腋淋巴结转移的临床研究

目的:探讨彩色多普勒超声(CDU)赋分诊断(HER-2、ER和PR)三阴性乳腺癌(TNBC)并腋淋巴结转移(ALNM)的临床应用价值。方法:回顾性分析2014年1月至2015年12月期间在青岛大学附属医院经CDU检查、手术和病理证实的863例女性乳腺癌患者的临床资料。根据临床病理学及免疫组化(的)检测结果(将其)分为TNBC组和非三阴性乳腺癌(NTNBC)组。对比分析两组乳腺癌的CDU征象,采用多因素非条件Logistic回归分析CDU赋分诊断与TNBC的相关性。将CDU描述性指标纳入赋分(0~15分),赋分10分者为NTNBC,8~10分为疑似TNBC,8分为倾向TNBC。结果:本组乳腺癌CDU诊断与病理符合率为90.96%(785/863),删除不能确定乳腺肿块性质78例(9.04%)。NBC组(105例)与NTNBC组(680例),肿块呈圆形或椭圆形(48.57%vs10.70%)、最大径5.0 cm(21.90%vs15.30%)、无微钙化灶(87.62%vs74.12%)、境界清楚(56.19%vs41.47%)、边缘无毛刺征(74.29%vs41.62%)、周边无高回声晕环(74.29%vs46.76%)、后方回声无衰减(72.38%vs46.76%)、ALNM(61.90%vs 37.35%)等比较有明显差异(P0.05),而在肿块数目、纵横径比、回声程度、回声分布和血流分级等则无明显差异(P0.05)。CDU赋分10分为NTNBC与病理符合率93.93%(680/724);8~10分疑似诊断TNBC符合率11.51%(16/139);8分倾向诊断TNBC符合率64.03%(89/139),TNBC总诊断符合率为75.54%(105/139)。在CDU诊断785例乳腺癌中,ALN肿大319例(40.64%),其中TNBC组61.90%(65/105),NTNBC组37.35%(254/680)。经多因素非条件Logistic回归分析结果显示:肿块呈圆形或椭圆形、边缘无毛刺征、周边无高回声晕环、后方回声无衰减和ALNM等与TNBC存在相关性(P0.05)。结论:乳腺貌似良性肿块并腋淋巴结肿大,CDU赋分8分,同时年龄≤50岁或未绝经,或有乳腺癌家族易感史者,可作为CDU诊断TNBC并ALNM重要参考指标,具有较高的临床应用价值。     

Clinicopathological features and prognosis of triple negative breast cancer in Kuwait: A comparative/perspective analysis

Aim

The aim of this study was to determine the incidence of TNBC in Kuwait, to analyze the clinicopathologic features and prognosis of this type of breast cancer, and compare it with reports from other regions of the world.

Background

Triple negative breast cancer (TNBC) is defined as a subtype that is negative for estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 (HER2). There is a growing evidence of the heterogeneity of such entity on the molecular level that may cause discrete outcomes.

Methods

We analyzed the clinicopathologic features of 363 TNBC cases which were diagnosed in Kuwait from July 1999 to June 2009. The disease-free survival (DFS) and overall survival (OS) were analyzed by Kaplan–Meier method. Comparison was done with reports from USA, Europe, Middle and Far East.

Results

Among 2986 patients diagnosed with breast cancer in Kuwait, 363 patients (12.2%) were TNBC. The median age was 48 years, 57.2% had lymph nodes (LN) metastasis, 56.9% were of grade III tumor and 41.9% had stage II disease. 81% developed recurrences and 75% of deaths occurred by 2.5 years after treatment. There is marked variation of clinicopathologic features according to country of patients’ cohort.

Conclusion

The incidence of TNBC in our study is similar to other studies. TNBC patients showed an early major recurrence surge peaking at approximately year 2.5. Regional variation of clinicopathologic features indicates a need for molecular studies to define underlying molecular features and its impact on survival.     

Human breast cancer associated fibroblasts exhibit subtype specific gene expression profiles

ABSTRACT: BACKGROUND: Breast cancer is a heterogeneous disease for which prognosis and treatment strategies are largely governed by the receptor status (estrogen, progesterone and Her2) of the tumor cells. Gene expression profiling of whole breast tumors further stratifies breast cancer into several molecular subtypes which also co-segregate with the receptor status of the tumor cells. We postulated that cancer associated fibroblasts (CAFs) within the tumor stroma may exhibit subtype specific gene expression profiles and thus contribute to the biology of the disease in a subtype specific manner. Several studies have reported gene expression profile differences between CAFs and normal breast fibroblasts but in none of these studies were the results stratified based on tumor subtypes. METHODS: To address whether gene expression in breast cancer associated fibroblasts varies between breast cancer subtypes, we compared the gene expression profiles of early passage primary CAFs isolated from twenty human breast cancer samples representing three main subtypes; seven ER+, seven triple negative (TNBC) and six Her2+. RESULTS: We observed significant expression differences between CAFs derived from Her2+ breast cancer and CAFs from TNBC and ER + cancers, particularly in pathways associated with cytoskeleton and integrin signaling. In the case of Her2+ breast cancer, the signaling pathways found to be selectively up regulated in CAFs likely contribute to the enhanced migration of breast cancer cells in transwell assays and may contribute to the unfavorable prognosis of Her2+ breast cancer. CONCLUSIONS: These data demonstrate that in addition to the distinct molecular profiles that characterize the neoplastic cells, CAF gene expression is also differentially regulated in distinct subtypes of breast cancer.     

三阴性乳腺癌靶向治疗相关信号通路及其临床应用

三阴性乳腺癌(triple negative breast cancer, TNBC)占全部乳腺癌病例的15%~20%,其雌激素受体、孕激素受体和人表皮生长因子受体2均为阴性表达,也是所有乳腺癌亚型中侵袭性和恶性程度较高的一种。TNBC还具有较高的复发风险和较差的预后特性。由于异质性高、临床特征复杂,化疗、放疗和手术切除等手段仍是当前TNBC治疗的主要方法。然而,严重的副作用、高复发风险和健康损伤等问题仍然不容忽视。随着TNBC基础研究的进展,越来越多的TNBC靶向治疗相关信号通路被揭示,而且其中有一部分已进入临床试验,为TNBC的治疗提供了充满希望和前景的分子靶点。此外,其中一些治疗靶点在TNBC精确分型和精准治疗的临床实践中发挥着重要的作用。本文对TNBC靶向治疗中经典的合成致死通路、PI3K/AKT/mTOR通路、PD-1/PD-L1免疫通路等信号通路及其临床试验进行了综述,同时介绍了近几年比较具有潜力的TNBC靶向治疗信号通路,包括肿瘤血管生成通路、多胺合成和分解代谢通路、SLC3A2/LAT1通路以及IGF-1/IGF-1R/FAK/YAP信号转导通路等。     

Comprehensive analysis of novel three-long noncoding RNA signatures as a diagnostic and prognostic biomarkers of human triple-negative breast cancer

Currently, traditional predictors of prognosis (tumor size, nodal status, progesterone receptor [PR], estrogen receptor [ER], or human epidermal growth factor receptor-2 [HER2]) are insufficient for precise survival prediction for triple-negative breast cancer (TNBC). Long noncoding RNAs (lncRNAs) have been observed to exert critical functions in cancer, including in TNBC. Nevertheless, systematically tracking expression-based lncRNA biomarkers based on the sequence data for the prediction of prognosis in TNBC has not yet been investigated. To ascertain whether biomarkers exist that can distinguish TNBC from adjacent normal tissue or nTNBC, we implemented a comprehensive analysis of lncRNA expression profiles and clinical data of 1097 BC samples from The Cancer Genome Atlas database. A total of 1510 differentially expressed lncRNAs in normal and TNBC samples were extracted. Similarly, 672 differentially expressed lncRNAs between nTNBC and TNBC samples were detected. The receiver operating characteristic curve analysis indicated that three upregulated lncRNAs (AC091043.1, AP000924.1, and FOXCUT) may be of strong diagnostic value for predicting the existence of TNBC in the training and validation sets (area under the curve (AUC > 0.85). Kaplan-Meier analysis demonstrated that the other three lncRNAs (AC010343.3, AL354793.1, and FGF10-AS1) were associated with the prognosis of TNBC patients (P < 0.05). We used the three overall survival (OS)-related lncRNAs to establish a three-lncRNA signature. Multivariate Cox regression analysis suggested that the three-lncRNA signature was a prognostic factor independent of other clinical variables ( P < 0.01) for predicting OS in TNBC patients that could be utilized to classify patients into high- or low-risk subgroups. Our results might provide efficient signatures for clinical diagnosis and prognostic evaluation of TNBC.     

IL-26 mediates epidermal growth factor receptor-tyrosine kinase inhibitor resistance through endoplasmic reticulum stress signaling pathway in triple-negative breast cancer cells

Triple-negative breast cancer (TNBC) has a poor prognosis compared to other breast cancer subtypes. Although epidermal growth factor receptor (EGFR) is overexpressed in TNBC, clinical trials with EGFR inhibitors including tyrosine kinase inhibitors (EGFR-TKI) in TNBC have heretofore been unsuccessful. To develop effective EGFR-targeted therapy for TNBC, the precise mechanisms of EGFR-TKI resistance in TNBC need to be elucidated. In this study, to understand the molecular mechanisms involved in the differences in EGFR-TKI efficacy on TNBC between human and mouse, we focused on the effect of IL-26, which is absent in mice. In vitro analysis showed that IL-26 activated AKT and JNK signaling of bypass pathway of EGFR-TKI in both murine and human TNBC cells. We next investigated the mechanisms involved in IL-26-mediated EGFR-TKI resistance in TNBC. We identified EphA3 as a novel functional receptor for IL-26 in TNBC. IL-26 induced dephosphorylation and downmodulation of EphA3 in TNBC, which resulted in increased phosphorylation of AKT and JNK against EGFR-TKI-induced endoplasmic reticulum (ER) stress, leading to tumor growth. Meanwhile, the blockade of IL-26 overcame EGFR-TKI resistance in TNBC. Since the gene encoding IL-26 is absent in mice, we utilized human IL-26 transgenic (hIL-26Tg) mice as a tumor-bearing murine model to characterize the role of IL-26 in the differential effect of EGFR-TKI in human and mice and to confirm our in vitro findings. Our findings indicate that IL-26 activates the bypass pathway of EGFR-TKI, while blockade of IL-26 overcomes EGFR-TKI resistance in TNBC via enhancement of ER stress signaling. Our work provides novel insights into the mechanisms of EGFR-TKI resistance in TNBC via interaction of IL-26 with its newly identified receptor EphA3, while also suggesting IL-26 as a possible therapeutic target in TNBC.Subject terms: Stress signalling, Cell death and immune response, Breast cancer     

Putative tumor suppressor cytoglobin promotes aryl hydrocarbon receptor ligand–mediated triple negative breast cancer cell death

Nearly 40 000 women die annually from breast cancer in the United States. Clinically available targeted breast cancer therapy is largely ineffective in triple negative breast cancer (TNBC), characterized by tumors that lack expression of the estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (Her2). TNBC is associated with a poor prognosis. Previous reports show that aryl hydrocarbon receptor (AhR) partial agonist 2-(4-amino-3-methylphenyl)-5-fluorobenzothiazole (5F 203) selectively inhibits the growth of breast cancer cells, including those of the TNBC subtype. We previously demonstrated that 5F 203 induced the expression of putative tumor suppressor gene cytoglobin (CYGB) in breast cancer cells. In the current study, we determined that 5F 203 induces apoptosis and caspase-3 activation in MDA-MB-468 TNBC cells and in T47D ER⁺ PR ⁺ Her2 ⁻ breast cancer cells. We also show that caspases and CYGB promote 5F 203–mediated apoptosis in MDA-MB-468 cells. 5F 203 induced lysosomal membrane permeabilization (LMP) and cathepsin B release in MDA-MB-468 and T47D cells. In addition, silencing CYGB attenuated the ability of 5F 203 to induce caspase-3/-7 activation, proapoptotic gene expression, LMP, and cathepsin B release in MDA-MB-468 cells. Moreover, 5F 203 induced CYGB protein expression, proapoptotic protein expression, and caspase-3 cleavage in MDA-MB-468 cells and in MDA-MB-468 xenograft tumors grown orthotopically in athymic mice. These data provide a basis for the development of AhR ligands with the potential to restore CYGB expression as a novel strategy to treat TNBC.     

不同分子分型乳腺癌患者血清IGFBP-3、Angptl-2表达水平及其与骨转移、预后的相关性

摘要 目的：分析不同分子分型乳腺癌患者血清胰岛素样生长因子结合蛋白3（IGFBP-3）、生成素养蛋白2（Angptl-2）表达水平及其与骨转移、预后的相关性。方法：选取2018年3月-2021年3月东南大学附属中大医院收治的128例乳腺癌骨转移患者进行研究,其中包括Luminal A型50例、42例Luminal B型（HER-2阴性）42例、HER-2过表达型16例、三阴性乳腺癌（TNBC）20例,并分析4种分子分型乳腺癌的临床病理特征,同时采用酶联免疫吸附法检测其血清IGFBP-3、Angptl-2表达水平;随访24个月后记录两组患者的预后情况,并采用多因素Logistic模型分析影响4种分子分型乳腺癌骨转移患者预后的独立危险因素,以及血清IGFBP-3、Angptl-2与不同分子分型乳腺癌骨转移患者预后的相关性。结果：Luminal A型、Luminal B型、HER-2过表达型、TNBC型TNM分期、淋巴结转移比较,差异有统计学意义（P＜0.05）。与Luminal A型、Luminal B型、TNBC型乳腺癌骨转移患者相比,HER-2过表达型乳腺癌骨转移患者的血清IGFBP-3表达水平较低,Angptl-2表达水平较高（P＜0.05）。Luminal A型、Luminal B型、HER-2过表达型、TNBC型乳腺癌骨转移患者的死亡率分别为13.46%、38.46%、23.08%、25.00%。多因素Logistic结果显示,TNM分期、淋巴结转移、血清IGFBP-3、Angptl-2均是影响不同分子分型乳腺癌骨转移患者预后的独立危险因素（P＜0.05）。血清IGFBP-3异常高表达提示4种分子分型乳腺癌骨转移患者的不良预后,而Angptl-2表达水平与4种分子分型乳腺癌的预后呈正相关性（P＜0.05）。针对不同分子分型乳腺癌骨转移患者的预后预测中,血清IGFBP-3、Angptl-2、IGFBP-3+Angptl-2均呈现AUC＞0.75。结论：血清IGFBP-3、Angptl-2可作为HER-2过表达乳腺癌骨转移患者的潜在生物标志物;同时还可根据血清IGFBP-3、Angptl-2表达水平预测不同分子分型乳腺癌骨转移患者的预后。     

GABA(A) Receptor Pi (GABRP) Stimulates Basal-like Breast Cancer Cell Migration through Activation of Extracellular-regulated Kinase 1/2 (ERK1/2)

Breast cancer is a heterogeneous disease comprised of distinct subtypes predictive of patient outcome. Tumors of the basal-like subtype have a poor prognosis due to inherent aggressiveness and the lack of targeted therapeutics. Basal-like tumors typically lack estrogen receptor-α, progesterone receptor and HER2/ERBB2, or in other words they are triple negative (TN). Continued evaluation of basal-like breast cancer (BLBC) biology is essential to identify novel therapeutic targets. Expression of the pi subunit of the GABA(A) receptor (GABRP) is associated with the BLBC/TN subtype, and herein, we reveal its expression also correlates with metastases to the brain and poorer patient outcome. GABRP expression in breast cancer cell lines also demonstrates a significant correlation with the basal-like subtype suggesting that GABRP functions in the initiation and/or progression of basal-like tumors. To address this postulate, we stably silenced GABRP in two BLBC cell lines, HCC1187 and HCC70 cells. Decreased GABRP reduces in vitro tumorigenic potential and migration concurrent with alterations in the cytoskeleton, specifically diminished cellular protrusions and expression of the BLBC-associated cytokeratins, KRT5, KRT6B, KRT14, and KRT17. Silencing GABRP also decreases phosphorylation of extracellular regulated kinase 1/2 (ERK1/2) in both cell lines and selective inhibition of ERK1/2 similarly decreases the basal-like cytokeratins as well as migration. Combined, these data reveal a GABRP-ERK1/2-cytokeratin axis that maintains the migratory phenotype of basal-like breast cancer. GABRP is a component of a cell surface receptor, thus, these findings suggest that targeting this new signaling axis may have therapeutic potential in BLBC.