Rho GTPase-activating protein 10 (ARHGAP10) is recognized as a tumor suppressor, yet the functional impact of its alternative splicing isoforms on breast cancer metastasis remains unclear. This study aimed to elucidate the role and regulatory mechanism of ARHGAP10 exon 21 skipping in breast cancer progression. Our research results indicate that in metastatic breast cancer cells, the full-length isoform ARHGAP10-L is downregulated, whereas the truncated ARHGAP10-S is upregulated. The RNA-binding protein HNRNPA0 directly binds to intron 21 of ARHGAP10 pre-mRNA, promoting exon-21 skipping and ARHGAP10-S production. Functionally, ARHGAP10-L and ARHGAP10-S exert opposing effects on breast cancer cell malignancy: ARHGAP10-L suppresses migration, invasion, and lung metastasis, whereas ARHGAP10-S promotes these aggressive phenotypes. Moreover, ARHGAP10-S exhibits enhanced binding to CDC42 and is associated with increased AKT phosphorylation. In a nude mouse model, HNRNPA0 drove lung metastasis by upregulating ARHGAP10-S. These findings establish the HNRNPA0-ARHGAP10 splicing axis as a key regulator of breast cancer metastasis, in which ARHGAP10-S promotes progression via the AKT pathway whereas ARHGAP10-L acts as a tumor suppressor, highlighting the therapeutic potential of targeting this splicing event to combat metastasis.

Hypoxia-inducible factor 1-alpha (HIF1A) is a core regulator of cellular adaptation to hypoxic environments and is extensively involved in various cancer processes. Although it is known that HIF1A produces two isoforms, HIF1A-L and HIF1A-S, through the alternative splicing of exon 14, the specific functional differences between these isoforms in cancer development and progression remain unclear, and the molecular mechanisms regulating this exon skipping event have yet to be elucidated. Clinical IHC and FISH analyses demonstrate that HIF1A-L expression is elevated in high-grade breast cancer tissues and correlates with malignant progression. Using transcriptomics and functional assays, we demonstrate that HIF1A-L enhances, while HIF1A-S inhibits, cancer cell proliferation, migration, and invasion. Mechanistically, through luciferase reporter and Western blot analyses, we found that HIF1A-L upregulates CXCR4 to activate the AKT pathway, whereas HIF1A-S antagonizes this axis. Furthermore, via CL-RIP, MS2-RIP, and RNA-pull down assays, we identify the RNA-binding protein HNRNPF as the key upstream regulator that specifically binds to a conserved G-rich sequence (gggaggtggaggttgcgatgagctgagatcagg) within intron 13 of HIF1A pre-mRNA to promote exon 14 retention and HIF1A-L production. Critically, in vivo metastasis assays in nude mice reveal that knockdown of HNRNPF or HIF1A-L suppresses lung metastasis, along with reduced CXCR4 in lung tissues and lower serum levels of the pro-metastatic cytokines IL-6 and CXCL12, whereas knockdown of HIF1A-S exacerbates metastasis. This study elucidates the HNRNPF/HIF1A-L/CXCR4/AKT axis as a central regulatory circuit controlling breast cancer metastasis and suggests that correcting the aberrant splicing of HIF1A may represent a novel therapeutic strategy for cancer.

Objective:To investigate the impact of circASH1L on subcutaneous tumor growth in nude mice with triple-negative breast cancer via the PI3K/AKT pathway. Methods:The study was conducted using bioinformatics and animal experimental verification methods. circASH1L levels in triple-negative breast cancer (TNBC) were analyzed using a dataset from the gene expression omnibus database. In the animal experiment part, nude mice were divided into shNC group, shcircASH1L-1 group, Oe-NC group, and Oe-circASH1L group. Each group was treated with corresponding circASH1L overexpression or knockdown and transplanted tumor modeling. Immunohistochemistry and western blot experiments were used to verify the effect of circASH1L on the growth of nude mouse transplanted tumors and the PI3K/AKT pathway. Results:A total of 43 circRNAs were significantly associated with TNBC, among which circASH1L was significantly highly expressed in TNBC. circASH1L-1 negatively regulates tumor volume, mass, expression rate of Ki67 cells, and PI3K/AKT pathway marker proteins. Conclusions:CircASH1L is a tumor promoter in TNBC. The expression level of circASH1L influences both the proliferation of TNBC cells and the growth of TNBC nude mice tumors by modulating the PI3K/AKT pathway.

Chemoresistance remains a crucial obstacle in breast cancer therapy. The mechanisms underlying chemoresistance need to be explored urgently and in depth. Breast cancer metastasis suppressor 1 like (BRMS1L), a core component of the Sin3A–histone deacetylase (HDAC) co-repressor complex, has been reported to suppress breast cancer metastasis through epigenetically regulating the Wnt signal pathway. However, whether BRMS1L could regulate chemosensitivity has not been explored. Herein, we found that higher BRMS1L expression was significantly correlated with increased chemotherapy sensitivity and better prognosis in patients receiving neoadjuvant chemotherapy. In vitro experiments confirmed that chemoresistant breast cancer cells exhibited decreased BRMS1L expression compared to chemosensitive cells. In vivo experiments in nude mice demonstrated that BRMS1L markedly strengthened the chemotherapy effects on xenografts. RNA sequencing (RNA-seq) was performed to elucidate the molecular mechanism underlying BRMS1L-mediated chemosensitivity. Bioinformatics analysis indicated that BRMS1L promotes chemotherapy sensitivity by regulating cellular autophagy. Furthermore, chemoresistant breast cancer cells exhibited elevated autophagy levels, and ectopic expression of BRMS1L significantly suppressed protective autophagy through downregulating ATG5. Collectively, these results revealed that BRMS1L enhances chemotherapy sensitivity via inhibiting protective autophagy. To our knowledge, this is the first study that showed that reduced BRMS1L expression is associated with poor response to neoadjuvant chemotherapy and unfavorable prognosis in breast cancer patients. Our findings reveal a novel role of BRMS1L in chemosensitivity and highlight its potential clinical application in the treatment of breast cancer.

Breast cancer is a leading cause of cancer-related deaths among women globally. It is imperative to explore novel biomarkers to predict breast cancer treatment response as well as progression. Here, we collected six breast cancer samples and paired normal tissues for high-throughput sequencing. By differential expression analysis, we found 1687 DEGs and identified the top 10 hub genes, including TOP2A, CDK1, BUB1B, KIF11, CCNA2, BUB1, CCNB1, KIF20A, DLGAP5 and CDC20. Univariate and multivariate Cox analyses on the METABRIC database and GSE96058 dataset demonstrated that KIF20A was an independent prognostic predictor for overall survival. KIF20A was positively correlated with cell cycle phases, including the cell cycle process, cycle G2 M phase transition and cell cycle DNA replication initiation. Single-cell analyses revealed that KIF20A was enriched in fibroblasts and endothelial within breast cancer stroma. Meanwhile, multidrug resistance (MDR) genes ABCB1, ABCC1 and ABCG2 were co-expressed with KIF20A in fibroblasts and endothelial cells within the stroma. MTABRIC database confirmed that high expression of KIF20A was positively correlated with treatment efficacy in patients with breast cancer. In conclusion, KIF20A could be served as a predictive biomarker for breast cancer prognosis and treatment outcomes. KIF20A may play a significant role by regulating cell cycle progression and modulating stromal progression in breast cancer. Our findings provided novel molecular insights that can guide personalized treatment strategies in breast cancer.

Background:Non-coding RNAs have received increasing attention in human tumors, with RNA interaction networks playing important roles in breast cancer. This study aims to explore novel circular RNAs and their mechanisms of biological function in breast cancer. Methods:Six HER2-positive breast cancer tissues and paired normal tissues were obtained for the whole transcriptome RNA sequencing. Differentially expressed (DE) circRNAs, miRNAs and mRNAs were identified and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of DERNAs were performed. DECircRNAs- DEmiRNAs- DEmRNAs networks were constructed and further verified by bioinformatics database analyses, luciferase assays and RIP assays. The expression level of circDOCK1 in breast cancer specimens was measured using qRT-PCR. Functional rescue experiments were conducted to explore the role of circDOCK1/miR-138-5p/GRB7 axis in breast cancer cells. The correlation of circDOCK1 expression and clinicopathologic features of 102 HER2 positive breast cancer patients was analyzed. Results:A total of 6960 DEmRNAs, 133 DE miRNAs and 1691 DE circRNAs were identified from HER2-positive breast cancer tissues and paracancerous tissues. Enrichment Analysis showed that the differential mRNAs were associated with cell division in biological processes and cell cycle and signaling pathways. GO and KEGG analysis demonstrated that DE circRNAs were mainly enriched in double-strand break repair, positive regulation of transcription by RNA polymerase II, nucleoplasma, nucleus, chromatin binding and protein binding. Forty networks of competing endogenous RNAs (ceRNAs) were constructed and circDOCK1/miR-138-5p/GRB7 axis was verified. Functional experiments revealed that the axis promotes migration and invasion of breast cancer cells. CircDOCK1 expression was elevated in breast cancer patients and correlated with adverse clinicopathologic parameters. Patients with high circDOCK1 level had poor outcomes. Conclusion:A novel circDOCK1/miR-138-5p/GRB7 axis promotes HER2 positive breast cancer metastasis and progression, providing a potential therapeutic target in the treatment of breast cancer.

Background Lung metastasis is a significant adverse predictor of prognosis in patients with breast cancer. Accurate estimation for the prognosis of patients with lung metastasis and population-based validation for the models are lacking. In the present study, we aimed to establish the nomogram to identify prognostic factors correlated with lung metastases and evaluate individualized survival in patients with lung metastasis based on SEER (Surveillance, Epidemiology, and End Results) database. Methods We selected 1197 patients diagnosed with breast cancer with lung metastasis (BCLM) from the SEER database and randomly assigned them to the training group ( n = 837) and the testing group ( n = 360). Based on univariate and multivariate Cox regression analysis, we evaluated the effects of multiple variables on survival in the training group and constructed a nomogram to predict the 1-, 2-, and 3-year survival probability of patients. The nomogram were verified internally and externally by Concordance index (C-index), Net Reclassification (NRI), Integrated Discrimination Improvement (IDI), Decision Curve Analysis (DCA), and calibration plots. Results According to the results of multi-factor Cox regression analysis, age, histopathology, grade, marital status, bone metastasis, brain metastasis, liver metastasis, human epidermal growth factor receptor 2 (HER2), estrogen receptor (ER), progesterone receptor (PR), surgery, neoadjuvant therapy and chemotherapy were considered as independent prognostic factors for patients with BCLM. The C-index in the training group was 0.719 and the testing group was 0.695, respectively. The AUC values of the 1-, 2-, and 3-year prognostic nomogram in the training group were 0.798, 0.790 and 0.793, and the corresponding AUC values in the testing group were 0.765, 0.761 and 0.722. The calculation results of IDI and NRI were shown. The nomograms significantly improved the risk reclassification for 1-, 2-, and 3-year overall mortality prediction compared with the AJCC 7th staging system. According to the calibration plot, nomograms showed good consistency between predicted and actual overall survival (OS) values for the patients with BCLM. DCA showed that nomograms had better net benefits at different threshold probabilities at different time points compared with the AJCC 7th staging system. Conclusions Nomograms that predicted 1-, 2-, and 3-year OS for patients with BCLM were successfully constructed and validated to help physicians in evaluating the high risk of mortality in breast cancer patients.

癌细胞通过调节代谢方式以提供大量的ATP和生物分子,来满足细胞生长、分裂和生存的需要.近年来,癌细胞中脂质代谢的变化受到越来越多的关注,因为脂肪酸不仅是膜基质的主要成分,也是ATP和次级信使的主要来源.乳腺癌细胞的脂质代谢受Warburg效应的调控,癌细胞合成大量脂肪酸并促进其生成膜磷脂来进行信号传导和癌细胞的日常代谢.脂肪酸合成酶FASN是脂肪酸从头合成的关键酶,越来越多的研究认为FASN过表达可促进乳腺癌细胞对治疗的抵抗.本综述中,我们重点介绍FASN诱导乳腺癌全身系统治疗抵抗方面的最新进展,并展望靶向抑制FASN为乳腺癌治疗增敏的潜在临床应用价值.

<p>Legend for Supplementary Figures S1-S6.</p>

Abstract Background Gynecomastia is a common condition in clinical practice. The present study aimed to review the clinical data of ER-positive gynecomastia patients treated by tamoxifen (TAM) versus surgery and discussed the clinical effects of the two treatment strategies. Method We retrospectively collected the clinical indicators of patients with unilateral or bilateral gynecomastia who received treatment at our hospital between April 2018 and December 2021. Depending on the treatment received, the patients were divided into TAM and surgery groups. Result A total of 170 patients were recruited, including 91 patients in TAM group and 79 patients in surgery group. The age of the patients differed significantly between the TAM and surgery groups (P < 0.01). The estrogen level was closer in patients with stable and progressive disease, but significantly different in patients of glandular shrinkage in TAM group (P < 0.01). The proportion of patients achieving stable disease was higher among those with clinical grade 1–2. Among patients classified as clinical grade 3, the proportion of patients achieving glandular shrinkage of the breast was higher after TAM treatment (P < 0.05). The age and length of hospital stay were significantly different in patients undergoing open surgery than minimally invasive rotary cutting surgery and mammoscopic-assisted glandular resection (P < 0.01). Patients had significantly different complications including mild postoperative pain, hematoma, nipple necrosis, nipple paresthesias and effusions among the surgery subgroups (all P < 0.05). The estrogen level and the type of surgery were significantly different between the surgical recurrence and non-recurrence subgroups (P < 0.05). The difference in the thickness of glandular tissues upon the color Doppler ultrasound also reached a statistical significance between the two groups (P = 0.050). An elevated estrogen level was a factor leading to TAM failure. Among surgical patients, the thickness of glandular tissues, estrogen level, and type of surgery performed were risk factors for postoperative recurrence (all P < 0.05). Conclusion Both treatment strategies can effectively treat gynecomastia, but different treatment methods can benefit different patients. TAM treatment is more beneficial than surgery for patients who cannot tolerate surgery, have a low estrogen level, and are clinical grade 1–2. Surgery treatment is better than TAM for patients of clinical grade 3. Different surgery options may lead to different complications. Patients with a greater glandular tissue thickness and a higher estrogen level were shown to have a higher risk of recurrence.

In recent years, breast cancer attracts more and more attention because of its high incidence. To explore the molecular functions and mechanisms, we performed RNA sequencing on the tumor tissues and their paired normal tissues from three breast cancer patients. By differential expression analysis, we found 3764 differentially expressed (DE) mRNAs, 5416 DE lncRNAs, and 148 DE circRNAs. Enrichment analysis suggested that the DE lncRNAs and DE circRNAs were enriched in mitochondria and nucleus, which indicated that they may participate in the vital metabolism directly or indirectly, such as fatty acid metabolism. Subsequently, the protein-protein interaction (PPI) network was constructed and we got 8 key proteins, of which the matrix metalloproteinase-9 (MMP9; degree 5) draws our attention. Based on the 38 up-regulated circRNAs and 14 down-regulated circRNAs, we constructed competing endogenous RNA (ceRNA) networks, from which the has-miR-6794-5p has been identified to enriched in the up-regulated network and correlated with the circNFIX directly. At this point, we presented that the circNFIX and MMP9 may play a significant role by regulating fatty acid metabolism in breast cancer.

Abstract Background: Ubiquitin-conjugating enzyme E2T (UBE2T), an E2 ubiquitin-conjugating enzyme, is verified to play an important role in the tumorigenesis of various cancers. By downregulation of BRCA1, UBE2T affects the growth of BC cells. However, the clinical significance of UBE2T is uncertain. To explore the role of UBE2T in breast cancer (BC). Methods: Firstly, we analyzed the correlations between UBE2T and clinicopathological characteristics of BC by the TCGA database. Next, Cox regression and Kaplan-Meier analysis were utilized for association between clinicopathological features and the overall survival (OS). Kaplan-Meier analysis and ROC plotter were used for survival analysis and drug sensitivity. Results：UBE2T was significantly upregulated in BC in comparison with para cancer tissue(p<0.001). The high expression of UBE2T is related to lager tumor sizes (p=0.006) and later clinical stages (p=0.001). UBE2T is verified to be correlated with ER, PR, HER2 status of BC. The expression level of UBE2T varies in different molecular subtypes of BC. Further research revealed that UBE2T mainly affected the recurrence-free time of BC in subgroups. Conclusions: Univariate analysis and multivariate analysis certified that UBE2T was an independent indicator for BC prognosis. UBE2T is proven to be a good biomarker for predicting the responds of BC patients to aromatase inhibitor (AI).

BACKGROUND Granulocytic sarcoma (GS) is a rare malignant tumor, and relapse is even rarer in the breast and dorsal spine following allogeneic hematopoietic stem cell transplantation. Currently, a standard treatment regimen is not available. CASE SUMMARY A rare case of GS of the right breast and dorsal spine after complete remission of acute myelogenous leukemia is reported here. A 55-year-old female patient presented with a palpable, growing, painless lump as well as worsening dorsal compressive myelopathy. She had a history of acute myelomonocytic leukemia (AML M4) and achieved complete remission after chemotherapy following allogeneic hematopoietic stem cell transplantation. Imaging examinations showed the breast lump and C7-T1 epidural masses suspected of malignancy. Histologic results were compatible with GS in both the right breast and dorsal spine, which were considered extramedullary relapse of the AML treated 4 years earlier. CONCLUSION A rare case of GS relapse following allogeneic hematopoietic stem cell transplantation and guidelines for treatment are discussed.

Background: Breast phyllodes tumor (PT), the rare fibroepithelial mammary tumor is composed by mesenchymal and epithelial components, only 1%~2% of which were accompanied by epithelial breast cancer. The incidence of non-invasive ductal carcinoma (DCIS) arising in benign PTs is even rarer. There is no certain treatment for it currently because of the low incidence of PT with DCIS. Case Description: Here, we report a 57-year-old female patient who was diagnosed with ductal carcinoma in situ arising in a benign phyllodes tumor of the breast. Mastectomy and sentinel lymph nodes biopsy were performed, and endocrine therapy was administrated to the patient. No recurrence was noted during follow-up. Conclusion: Since PTs are prone to recurrence, adjuvant therapy should be considered comprehensively by the histopathological results after surgical treatment. In addition, regular follow-up is recommended.

乳腺癌是全球女性发病率最高的恶性肿瘤, 2020年全球乳腺癌新发病例高达226万例,超过了肺癌成为全球第一大肿瘤,严重威胁着女性健康[1].乳腺癌是高度异质性的疾病,第2代测序技术证实,在93个编码蛋白质的乳腺癌基因中携带可能的驱动突变[2].因此,现行的基于乳腺癌临床病理特征以及雌激素受体(estrogen receptor,ER)、孕激素受体(progesterone receptor,PR)及人表皮生长因子受体 2 (human epidermal growth factor receptor 2, HER2)的表达制定综合方案,并非适用于所有的乳腺癌患者.传统的二维细胞培养和人源性肿瘤异种移植模型为乳腺癌的研究探索提供了大量的材料,但由于实验技术缺陷、培养周期长等因素难以满足深入的研究要求.因此,建立一种能够准确模拟人体肿瘤特征的实验模型成为迫切需要.近年来,通过优化各种培养条件建立的乳腺癌类器官模型,由于其取材方便、培养周期短和遗传稳定等特点,能够最大程度的还原乳腺癌细胞在体内的组织和分子特性,重现部分乳腺功能[3],有望在乳腺癌发病机制研究、个体化治疗、新药开发等领域发挥重要作用.

Gynecomastia is the most common benign disease in males with an increasing prevalence in recent years. It may cause local pain and psychological disorders. The vacuum-assisted breast biopsy system has been reported to be a novel surgical approach for the treatment of gynecomastia. However, there are little detailed reports comparing the curative effect between traditional surgery and vacuum-assisted breast biopsy for gynecomastia. Besides, there was little study which compared the application of two different systems for the treatment of gynecomastia. Our study aimed to investigate the effectiveness of vacuum-assisted breast biopsy systems for patients with gynecomastia. We retrospectively reviewed 83 patients with gynecomastia between January 2015 and December 2019. Open surgery was performed in 56 patients, and vacuum-assisted breast biopsy was performed in 27 patients. The characteristics of patients as well as the curative effects between the two groups were analyzed. The two vacuum-assisted breast biopsy systems (Mammotome and Encor) were performed for the patients with gynecomastia. The efficacy, safety, complications, and patient satisfactions were recorded during postoperative follow-up periods. Compared with the open surgery group, the vacuum-assisted breast biopsy group had significantly smaller scar sizes left after the operation (5.5 ± 1.3 cm vs 0.8 ± 0.2 cm, p < 0.001), and shorter hospital stay time (5.5 ± 2.4 ds vs 3.1 ± 1.6 ds, p < 0.001). Patients in vacuum-assisted breast biopsy group had a better cosmetic outcome than those in open surgery group. There were no statistically significant differences between the two vacuum-assisted breast biopsy systems according to the mean age, the mean operation time, sites, or grade. In addition, no serious complications were observed in vacuum-assisted breast biopsy group. All the patients recovered well and were satisfied with the cosmetic outcomes. The vacuum-assisted breast biopsy system can be used as a feasible and minimally invasive approach for the treatment of gynecomastia. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Background With the extensive application of autologous fat grafting (AFG) to the breasts, postoperative complications such as breast lumps attract high attention. Breast lumps greatly reduce patient satisfaction and bring mental stress. However, there are few detailed reports about minimally invasive treatment strategies for breast lumps after AFG. Our study aimed to investigate the effectiveness of the vacuum-assisted breast biopsy (VABB) system for patients with lumps after AFG. Materials and Methods We retrospectively reviewed 37 patients with breast lumps between April 2015 and January 2019. The characteristics of patients and breast lumps were analyzed. Breast lumps were classified into four types, including cystic, solid, complex and calcification. The vacuum-assisted breast biopsy (Mammotome and Encor) was performed for the patients with lumps after AFG. The efficacy, safety, complications and patient satisfactions were recorded during postoperative follow-up periods. Results Under the guidance of ultrasound, the breast lumps could be thoroughly and accurately excised by the vacuum-assisted biopsy system. No patient experienced breast infections or major complications requiring treatment. Hematoma was observed in only 2 patients and gradually resolved without any special management. With a median follow-up of 29 months, no recurrence was observed. Furthermore, there were no statistical differences in duration of the procedures and complications between the two VABB systems. All the patients recovered well and were satisfied with the cosmetic outcome. Conclusion The vacuum-assisted breast biopsy system can be used as an effective and minimally invasive approach for the surgical management of lumps after AFG. Level of Evidence IV This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

目的:研究肿瘤出芽与乳腺癌临床病理特征、肿瘤浸润淋巴细胞(TILs)以及患者预后的关系.方法:收集2012年1月～2016年12月于暨南大学附属第一医院行手术治疗的178例乳腺癌患者资料及肿瘤组织切片,显微镜下观察乳腺癌组织病理切片中肿瘤出芽和肿瘤浸润淋巴细胞水平,x2检验分析肿瘤出芽水平与乳腺癌患者临床病理特征和TILs的关系,Log-rank检验分析肿瘤出芽水平与乳腺癌患者无病生存期和总生存期的关系.结果:高肿瘤出芽组患者淋巴结阳性数目多、组织性分级高、脉管癌栓更多;肿瘤出芽数较多的患者TILs的水平较低,而肿瘤出芽数较少的患者TILs水平较高;高肿瘤出芽患者比低肿瘤出芽患者预后较差.结论:乳腺癌肿瘤出芽水平与恶性程度高的临床病理指标密切相关,肿瘤出芽水平与TILs水平呈负相关,是影响乳腺癌预后的重要因素.

Long non-coding RNA is a kind of endogenous RNA with more than 200 nucleotides in length that cannot be coded to proteins.The dysregulation of lncRNAs have been observed in various malignancies.Aberrant ex-pression of lncRNAs levels are closely associated with various malignant biological processes,including carcinogenesis, progression,drug resistance,metastasis and metabolism,through transcriptional,posttranscriptional and epigenetic regu-lation of different genes.Nowadays,breast cancer has become the most common malignant tumor and the leading course of cancer-related death among women in China.lncRNAs are significant in a series of biological processes and may be served as a potential biomarker in breast cancer.Based on the latest research evidence,the present review will summa-rize the role of lncRNA in breast cancer,which will provide a better understanding of lncRNAs in preventing,diagnos-ing,and treating breast cancer.