luminal-A Breast Cancer Research Articles

Abstract B06: Similarity in breast cancer diversity: A novel target for breast cancer therapy

Abstract There is cancer; there is breast cancer (BCA); then there is luminal-A BCA, luminal-B BCA, HER2-enriched BCA, triple-negative/basal-like BCA; and, yes, normal-like BCA. In each of these distinct forms of BCA, unique targetable vulnerabilities in the tumors’ biologic makeup have been exploited, in many cases rather successfully, for drug development. To be sure, this divide and conquer approach has worked to a point. However, limitations include the targeting of proteins that, while overexpressed in a particular cancer, are also found in normal cells and the fact that many of these targets appear in very small subsets of BCA and thus the need to extensively type each cancer and to treat each individual cancer in a very specific fashion. Here, we are proposing another time-proven success strategy: unite and govern. We are targeting a commonality within the diverse array of breast cancers: the ASPH gene and its product, human aspartyl (asparaginyl) β-hydroxylase (HAAH). This enzyme has been detected specifically in more than 90% of all breast cancers by immunostaining (IHC/ICC), gene expression analysis (RT-PCR), and immunoassay (serum ELISA). When normal cells are transfected with ASPH, as HAAH is produced and translocated to the surface, they undergo malignant transformation and when ASPH is silenced in tumor cells, they revert to normal phenotype. Moreover, HAAH has a pivotal role in cell proliferation, motility, and invasiveness, the hallmarks of cancer. Also, the HAAH level in tumors is directly correlated with severity of disease and patient survival. HAAH is an embryonic enzyme silenced post-birth, and as such it is a self-protein. We have designed, produced, tested, manufactured, and are now clinically evaluating an HAAH-targeting therapeutic vaccine. This vaccine has effectively overcome self-tolerance and has the ability to become the ultimate cancer immunotherapy agent. In syngeneic rodent models, it is highly immunogenic, and in challenge models using various cell lines, including BCA lines, it has been effective in preventing tumor growth (up to 100%), reducing tumor size (over 90%), controlling metastasis (to over 80%), and increasing survival (to 100%). Even using a full human dose (40 μg total protein), no animal toxicities were reported (FDA-reviewed protocol). The vaccine is introduced intradermally using 3M’s hollow microneedle array device. In the ongoing phase I clinical trial, to date there have been no adverse complications or side effects reported. Furthermore, the vaccine has been demonstrated to be significantly immunogenic. We are now planning phase 2 clinical trials and propose to use this therapeutic cancer vaccine in HAAH-positive breast cancer patients (as high as 90% of all breast cancers). We intend to extend the clinical trials ultimately to all HAAH-positive cancer patients. Citation Format: Michael S. Lebowitz, Steven Fuller, Hossein A. Ghanbari. Similarity in breast cancer diversity: A novel target for breast cancer therapy [abstract]. In: Proceedings of the AACR Special Conference: Advances in Breast Cancer Research; 2017 Oct 7-10; Hollywood, CA. Philadelphia (PA): AACR; Mol Cancer Res 2018;16(8_Suppl):Abstract nr B06.

Metabolic profiles of triple-negative and luminal A breast cancer subtypes in African-American identify key metabolic differences.

Breast cancer, a heterogeneous disease with variable pathophysiology and biology, is classified into four major subtypes. While hormonal- and antibody-targeted therapies are effective in the patients with luminal and HER-2 subtypes, the patients with triple-negative breast cancer (TNBC) subtype do not benefit from these therapies. The incidence rates of TNBC subtype are higher in African-American women, and the evidence indicates that these women have worse prognosis compared to women of European descent. The reasons for this disparity remain unclear but are often attributed to TNBC biology. In this study, we performed metabolic analysis of breast tissues to identify how TNBC differs from luminal A breast cancer (LABC) subtypes within the African-American and Caucasian breast cancer patients, respectively. We used High-Resolution Magic Angle Spinning (HR-MAS) 1H Nuclear magnetic resonance (NMR) to perform the metabolomic analysis of breast cancer and adjacent normal tissues (total n=82 samples). TNBC and LABC subtypes in African American women exhibited different metabolic profiles. Metabolic profiles of these subtypes were also distinct from those revealed in Caucasian women. TNBC in African-American women expressed higher levels of glutathione, choline, and glutamine as well as profound metabolic alterations characterized by decreased mitochondrial respiration and increased glycolysis concomitant with decreased levels of ATP. TNBC in Caucasian women was associated with increased pyrimidine synthesis. These metabolic alterations could potentially be exploited as novel treatment targets for TNBC.

Differentiation Between Luminal-A and Luminal-B Breast Cancer Using Intravoxel Incoherent Motion and Dynamic Contrast-Enhanced Magnetic Resonance Imaging

The study aimed to investigate whether intravoxel incoherent motion (IVIM) and dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) can differentiate luminal-B from luminal-A breast cancer MATERIALS AND METHODS: Biexponential analyses of IVIM and DCE MRI were performed using a 3.0-T MRI scanner, involving 134 patients with 137 pathologically confirmed luminal-type invasive breast cancers. Luminal-type breast cancer was categorized as luminal-B breast cancer (LBBC, Ki-67 ≧ 14%) or luminal-A breast cancer (LABC, Ki-67 < 14%). Quantitative parameters from IVIM (pure diffusion coefficient [D], perfusion-related diffusion coefficient [D*], and fraction [f]) and DCE MRI (initial percentage of enhancement and signal enhancement ratio [SER]) were calculated. The apparent diffusion coefficient (ADC) was also calculated using monoexponential fitting. We correlated these data with the Ki-67 status. The D and ADC values of LBBC were significantly lower than those of LABC (P = 0.028, P = 0.037). The SER of LBBC was significantly higher than that of LABC (P = 0.004). A univariate analysis showed that a significantly lower D (<0.847 x 10-3 mm2/s), lower ADC (<0.960 × 10-3 mm2/s), and higher SER (>1.071) values were associated with LBBC (all P values <0.01), compared to LABC. In a multivariate analysis, a higher SER (>1.071; odds ratio: 3.0099, 95% confidence interval: 1.4246-6.3593; P = 0.003) value and a lower D (<0.847 × 10-3 mm2/s; odds ratio: 2.6878, 95% confidence interval: 1.0445-6.9162; P = 0.040) value were significantly associated with LBBC, compared to LABC. The SER derived from DCE MRI and the D derived from IVIM are associated independently with the Ki-67 status in patients with luminal-type breast cancer.

The Clinicopathological Factors Associated with Disease Progression in Luminal A Breast Cancer and Characteristics of Metastasis: A Retrospective Study from A Single Center in China.

This study investigated the clinicopathological factors associated with outcomes in patients with Luminal A breast cancer. Retrospective analysis of the association of clinicopathological factors and breast cancer outcome in 421 patients with newly-diagnosed Luminal-A breast cancer that were enrolled from January 2008 to December 2014. Clinicopathological data were analyzed to validate the relationship with disease-free survival (DFS) and overall survival (OS). Kaplan-Meier curves and log-rank tests were used to analyze the value of clinicopathological factors (tumor size, node status and lymphovascular invasion), and subsequent Cox regression analysis revealed significant prognostic factors. With a median of 61 months follow-up, the 5-year DFS and 5-year OS rate were 98.3% and 99.3%. Cox multivariate regression analysis showed that clinical anatomic stage, tumor size, status of lymph nodes, lymphovascular invasion and systemic treatment are strong prognostic factors for clinical outcome in patients with Luminal-A breast cancer. Of all 413 patients with stage I-III breast cancer, 14 presented with metastasis (3.4%) during the follow up. Bone (6/14, 42.9%) was the most common site of metastasis followed by liver (5/14, 35.7%) and lung (4/14, 28.6%). The median survival time after metastasis was 20.4 months. Of all the sites of distant metastasis, liver metastasis was the only factor that affected survival time after metastasis (χ2=6.263, p=0.012). Patients with Luminal A breast cancer have excellent outcomes. Liver metastasis is an important factor compressing the survival time after distant metastasis presents.

Abstract 5552: Differential proteomic responses of luminal-A and basal-like breast cancer cell lines during growth inhibition induced by co-culture with agarose encapsulated murine renal adenocarcinoma (RENCA) cells

Abstract The existence of a tumor growth regulatory network that is conserved across tumor types and species has been hypothesized. Our evidence for this regulatory network is derived from studies demonstrating tumor growth inhibition by agarose encapsulated cancer cells (cancer macrobeads). The ability of encapsulated, thus growth restricted cells, to inhibit freely growing cancer cells has been shown when the encapsulated cells are murine (RENCA) or human (e.g., J82). Clinical trials are underway (NCT01053013, NCT02046174). We have shown that RENCA macrobeads release &amp;gt;10 known tumor inhibitory proteins targeting several signaling pathways including Akt/PI3. To better understand the mechanism(s) of growth inhibition, we used a systems biology approach to identify protein profiles and interactions from macrobead-treated human breast carcinoma cell lines that are either mildly aggressive (MCF7) or highly aggressive (MDA-MB231 [MDA]). Target cells were co-cultured with RENCA macrobeads or left untreated for 5 days. Lysates of target cells were prepared in Laemmli buffer, frozen, and sent for MS/MS. Samples were run in duplicate, protein intensities normalized, and the treated/untreated ratio log10 transformed to get a normal distribution. Protein profiles were analyzed using Key Pathway Analysis (KPA) and Metacore software. Growth inhibition by RENCA macrobeads was confirmed (MCF7 25%; MDA 57%). KPA of MCF7 proteins showed upregulation of ubiquitin pathways involved in degradation of misfolded proteins. Stress induced apoptosis and DNA damage (which correlates with the growth inhibition) were the top 2 pathways upregulated using Pathway and Process Enrichment analysis. This is in line with a strong epigenetic gene deregulation (Metacore). Transcription Factor and Network analyses show upregulation of CREB, A2MR and ATF3 networks. Also, Regulated Network analysis suggests a role for A2MR or RAGE in the inhibition of proliferation and increased apoptosis. Macrobead-treated MDA cells showed no significant changes using KPA. Pathway, Network and Process enrichment (Metacore) showed 5 of the top 10 upregulated processes were associated with cytoskeleton remodeling. Cell cycle and protein folding processes were also upregulated. Transcription factors associated with this response were similar to that of MCF7. RAGE again appears in many of the pathways as indicated by Network and Regulated Network analysis. The protein response to RENCA macrobeads differs for these 2 cell lines. Proteins related to apoptosis are upregulated in the MCF7 cells whereas MDA have a preference for cytoskeleton remodeling. These data support the hypothesis that distinctive tumors may respond differently to RENCA macrobead exposure at both an epigenetic and protein level, nonetheless resulting in growth inhibition. Citation Format: Melissa A. Laramore, Peter James, Prithy C. Martis, Atira Dudley, Lawrence S. Gazda, Carl A. Borrebaeck, Barry H. Smith. Differential proteomic responses of luminal-A and basal-like breast cancer cell lines during growth inhibition induced by co-culture with agarose encapsulated murine renal adenocarcinoma (RENCA) cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5552. doi:10.1158/1538-7445.AM2017-5552

Network Analysis Reveals A Signaling Regulatory Loop in the PIK3CA-mutated Breast Cancer Predicting Survival Outcome

Mutated genes are rarely common even in the same pathological type between cancer patients and as such, it has been very challenging to interpret genome sequencing data and difficult to predict clinical outcomes. PIK3CA is one of a few genes whose mutations are relatively popular in tumors. For example, more than 46.6% of luminal-A breast cancer samples have PIK3CA mutated, whereas only 35.5% of all breast cancer samples contain PIK3CA mutations. To understand the function of PIK3CA mutations in luminal A breast cancer, we applied our recently-proposed Cancer Hallmark Network Framework to investigate the network motifs in the PIK3CA-mutated luminal A tumors. We found that more than 70% of the PIK3CA-mutated luminal A tumors contain a positive regulatory loop where a master regulator (PDGF-D), a second regulator (FLT1) and an output node (SHC1) work together. Importantly, we found the luminal A breast cancer patients harboring the PIK3CA mutation and this positive regulatory loop in their tumors have significantly longer survival than those harboring PIK3CA mutation only in their tumors. These findings suggest that the underlying molecular mechanism of PIK3CA mutations in luminal A patients can participate in a positive regulatory loop, and furthermore the positive regulatory loop (PDGF-D/FLT1/SHC1) has a predictive power for the survival of the PIK3CA-mutated luminal A patients.

IODNE: An integrated optimization method for identifying the deregulated subnetwork for precision medicine in cancer.

Subnetwork analysis can explore complex patterns of entire molecular pathways for the purpose of drug target identification. In this article, the gene expression profiles of a cohort of patients with breast cancer are integrated with protein‐protein interaction (PPI) networks using, simultaneously, both edge scoring and node scoring. A novel optimization algorithm, integrated optimization method to identify deregulated subnetwork (IODNE), is developed to search for the optimal dysregulated subnetwork of the merged gene and protein network. IODNE is applied to select subnetworks for Luminal‐A breast cancer from The Cancer Genome Atlas (TCGA) data. A large fraction of cancer‐related genes and the well‐known clinical targets, ER1/PR and HER2, are found by IODNE. This validates the utility of IODNE. When applying IODNE to the triple‐negative breast cancer (TNBC) subtype data, we identified subnetworks that contain genes such as ERBB2, HRAS, PGR, CAD, POLE, and SLC2A1.

Microenvironmental networks promote tumor heterogeneity and enrich for metastatic cancer stem-like cells in Luminal-A breast tumor cells.

The roles of the tumor microenvironment (TME) in generating intra-tumoral diversity within each specific breast cancer subtype are far from being fully elucidated. In this study, we exposed Luminal-A breast cancer cells in culture to combined “TME Stimulation”, representing three typical arms of the breast TME: hormonal (estrogen), inflammatory (tumor necrosis factor α) and growth-promoting (epidermal growth factor). In addition to enriching the tumor cell population with CD44+/β1+ cells (as we previously published), TME Stimulation selected for CD44+/CD24low/− stem-like cells, that were further enriched by doxorubicin treatment and demonstrated high plasticity in vitro and in vivo. Knock-down experiments revealed that CD44 and Zeb1 regulated CD24 and β1 expression and controlled differently cell spreading and formation of cellular protrusions. TME-enriched CD44+/CD24low/− stem-like cells promoted dissemination to bones and lymph nodes, whereas CD44+/β1+ cells had a low metastatic potential. Mixed co-injections of TME-enriched CD44+/CD24low/− and CD44+/β1+ sub-populations generated metastases populated mostly by CD44+/CD24low/−-derived cells. Thus, combined activities of several TME factors select for CD44+/CD24low/− stem-like cells that dictate the metastatic phenotype of Luminal-A breast tumor cells, suggesting that therapeutic modalities targeting the TME could be introduced as a potential strategy of inhibiting the detrimental stem-like sub-population in this disease subtype.

PV-0512: Accelerated partial breast irradiation for Luminal-A breast cancer: analysis from a phase 3 trial

PV-0512: Accelerated partial breast irradiation for Luminal-A breast cancer: analysis from a phase 3 trial

Abstract 121: The EGFR gatekeeper mutation T790M is present in selected patients with early breast cancer

Abstract Introduction. The epidermal growth factor receptor (EGFR) is one of the major oncogenes in a variety of human cancers including breast cancer (BC). While EGFR overexpression and/or amplification has been shown to occur frequently in human breast cancer, EGFR-activating mutations are suggested to be rare if not absent. Thus, the aim of our study was to examine the presence of somatic EGFR gene mutations in a group of norwegian patients diagnosed with early breast cancer. Patients. We investigated tumor biopsies obtained from 132 patients with early breast cancer treated at the Akershus University Hospital (University of Oslo) in 2007/2008. The majority of patients (n = 82) belonged to the luminal- A/B subtypes of BC. In addition, 33 patients were classified as HER-2 positive patients (IHC 3+ or amplification verified by FISH) and 17 as triple-negative breast cancer (TNBC) patients. Methods. Briefly, DNA was extracted from formalin-fixed, paraffin-embedded (FFPE) tumor tissue using QIAsymphony DSP DNA Mini Kit (Qiagen Inc.). The extraction of DNA was performed by fully automated purification using the QIAsymphony SP/AS system. EGFR-mutations were detected using a Therascreen EGFR RGQ PCR kit according to the manufacturer´s protocol. We tested for point mutations in exon 18 (G719A/C/S), in-frame deletions in exon 19 (ΔE746-A750), in-frame insertions as well as point mutations in exon 20 (S768I, T790M) and a point mutation in exon 21 (L858R). Results. In the present study we detected three individuals with an EGFR-T790M mutation. Two of the patients were diagnosed with TNBC, while the disease of the third patient was classified as a luminal-A type breast cancer. Interestingly, the T790M-mutation was present in the biopsies obtained during surgery before adjuvant therapy was initiated. No anti-cancer therapy has been given prior to surgery. According to our best knowledge, the T790M-”TKI-resistance-mutation” has not been detected in early breast cancer previously. This finding contrasts the observations made in lung cancer patients where the T790M mutation in general is a classical second mutation causing drug resistance during ongoing TKI-therapy. No other EGFR-mutations were detected in our patients. Conclusion: To our best knowledge we provide first evidence for the presence of the EGFR-T790M-mutation in human early breast cancer. Thus, treatment with novel EGFR-targeting cancer drugs (like “pan-HER-inhibitors”) affecting also T790M-mutated-EGFR may be worthwhile to be tested in highly selected breast cancer patients. Citation Format: Vahid Bemanian, Torill Sauer, Joel Touma, Bjørn A. Lindstedt, Ying Chen, Hilde P. Ødegård, Ida R. Bukholm, Jürgen Geisler. The EGFR gatekeeper mutation T790M is present in selected patients with early breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 121. doi:10.1158/1538-7445.AM2015-121

Abstract P4-05-04: Proteomic patterns unravel a new luminal-A breast cancer molecular subgroup with prognostic value

Abstract Background: Breast cancer is a heterogeneous disease including a variety of entities with different genetic background. Estrogen receptor-positive, HER2 negative tumors (ER+) usually have a favorable outcome, although some patients eventually relapse, which suggests some heterogeneity within this category. In the last years, proteomic approaches have been incorporated to the study of clinical samples as a way to complement the information provided by gene analysis. Shotgun proteomics allows measuring over 1,000 proteins in clinical samples. In the present study, we combined genomic and proteomic techniques to characterize a set of breast tumors. Methods: The study population consisted of 102 patients with lymph-node positive breast cancer who had received anthracycline-based adjuvant chemotherapy. Protein extracts from FFPE samples were prepared in 2% SDS buffer and digested with trypsin. SDS was removed from digested lysates, and resulting peptides were analyzed in an Orbitrap Velos. Protein abundance was calculated on the basis of the normalized spectral protein intensity (LFQ intensity) using MaxQuant. A prognostic protein signature was built. Findings were verified using whole genome gene expression data from 1,141 patients included in public repositories. To this purpose, the protein signature was converted to a gene signature. Data analysis was done using MeV, BRBArray Tools, R and Cytoscape software suites and Uniprot (http://www.uniprot.org/) and DAVID (http://david.abcc.ncifcrf.gov) webtools. Results: We identified 3,000 protein groups in FFPE breast cancer samples and selected 1,000 that were identified at least in 75% of the samples. Significance Analysis for Microarrays analysis revealed 224 protein groups differentially expressed between ER+ and triple-negative (TN) samples (False Discovery Rate set at &amp;lt;0.001). Hierarchical clustering analyses of protein expression showed that some ER+ samples had a protein expression profile similar to that of TN samples: patients with TN-like tumors had a clinical outcome similar to those with TN disease. Gene ontology analyses unraveled a reduced expression of proteins related with cellular adhesion in the TN-like and the TN groups. A TN-like predictive protein signature was built, converted to a gene signature and evaluated in the whole-genome expression data. The signature had prognostic value in patients with luminal-A breast cancer. This prognostic information was independent from that provided by standard genomic tests for breast cancer, such as MammaPrint, OncoType Dx and the 8-gene Score. Conclusions: Proteomic profiling showed that cellular adhesion is a differential process between ER+ and TN breast cancer, and is reduced in the TN tumors. A group of ER+ breast tumors with reduced cellular adhesion was identified (TN-like). Patients with this luminal-A, TN-like breast cancer type had a poor outcome. This prognostic information was complementary to that offered by genomic tests such as OncoType. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-05-04.

Triple Negative Breast Cancers Have a Reduced Expression of DNA Repair Genes

DNA repair is a key determinant in the cellular response to therapy and tumor repair status could play an important role in tailoring patient therapy. Our goal was to evaluate the mRNA of 13 genes involved in different DNA repair pathways (base excision, nucleotide excision, homologous recombination, and Fanconi anemia) in paraffin embedded samples of triple negative breast cancer (TNBC) compared to luminal A breast cancer (LABC). Most of the genes involved in nucleotide excision repair and Fanconi Anemia pathways, and CHK1 gene were significantly less expressed in TNBC than in LABC. PARP1 levels were higher in TNBC than in LABC. In univariate analysis high level of FANCA correlated with an increased overall survival and event free survival in TNBC; however multivariate analyses using Cox regression did not confirm FANCA as independent prognostic factor. These data support the evidence that TNBCs compared to LABCs harbour DNA repair defects.

Abstract 5003: The functional interplay between activated Stat5 and Stat3 in breast cancer formation and progression.

Abstract Cancer cells with stem cell-like characteristics are defined as cancer stem cells (CSCs) that are proposed to drive tumor initiation, progression and recurrence. Recent studies show the significance of epithelial-mesenchymal transition (EMT) in the induction of CSCs and their metastatic dissemination. Growth and colonization of the migrating stem cells require their re-differentiation and the reversal of EMT, mesenchymal to epithelial transition (MET), at the site of distant metastasis. We study the functional interplay between two related transcription factors, signal transducer and activator of transcription 5 (Stat5) and Stat3 in the genetic and epigenetic regulation of non-metastatic and metastatic breast cancer cells. Simultaneous activity of both Stats is observed in high percentage of human luminal breast cancers. In both normal and malignant cells, Stat5 activity is associated with cell proliferation, differentiation and survival, however, its activity in tumors is defined as a good prognostic marker due to response to antiestrogen therapy. During postlactational gland regression, called involution, P-Stat3 regulates lysosomal-mediated cell death, which is in contrast to its strong oncogenic function during mammary tumorigenesis. To gain insights into oncogenic functions of Stat5, we established a new mouse model which allows the investigation of genetic and epigenetic contributions in the initiation and progression of mammary tumors. In this model we used genetic manipulation of mammary stem cells (MaSCs) derived from primary tissue and the reconstitution of functional epithelium through transplantation into cleared fat pads to generate transgenic glands. The persistent activation of Stat5 during post-lactational involution resulted in the induction of non-metastatic adenocarcinomas, resembling human luminal-A breast cancer subtype. These tumors could be serially transplanted in BALB/c wild-type recipient mice and contain a small fraction of Lin-CD24lowCD44high tumor initiating cells. Both primary and secondary tumors show simultaneous activity of Stat5 and Stat3 and express estrogen and progesterone receptors (ER+PR+) and their downstream target genes like amphiregulin, RANKL and Wnt4. We performed microarray gene expression analysis of tumor tissues and P-Stat5 transgenic grafts and studied potential target genes, which can explain Stat5 oncogenic functions in tumor formation in contrast to its suppression effects on tumor cell migration. We are now studying the relative contribution of Stat5 and Stat3 in the regulation of metastatic process and transient EMT-MET changes. Our tumor model allows the identification of new genetic and epigenetic biomarkers, which are necessary for a better diagnosis and treatment of breast cancer and the prediction of invasive relapse. Citation Format: Vida Vafaizadeh, Bernd Groner. The functional interplay between activated Stat5 and Stat3 in breast cancer formation and progression. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5003. doi:10.1158/1538-7445.AM2013-5003

A prospective cohort study on hormone-dependant metastatic breast cancer treated with Shuganyishen formula

Objective Observing the effect of Shuganyishen formula on hormone-dependant metastatic breast cancer and the mechanisms of anti-resistance and endocrine therapy. Methods 226 ITT cases were divided into two cohorts by treatment. 126 cases were treated with the combination of Shuganyishen formula and standard western medicine including endocrine therapy, and 100 cases were treated with western medicine alone. The end points were time to progress (TTP) and overall survival and quality of life (QOF). The effect of Shuganyishen formula on growth of TAM-resistance cell line in vitro was observed. Results There were statistical significances of median TTP between combination and western medicine in Luminal A (ER+ /PR+, HER2-) as well as Luminal B (ER+/PR+, HER2+), but no significances of OS; The extent and incidence of hot flush, sweat and syndromes related to bone were decreased significantly in combination treated group. Shuganyishen formula could reverse resistance to TAM. Conclusion Patients with LuminalA and LuminalB metastatic breast cancer benefit from Shuganyishen formula, which maybe related with its reverse resistance to endocrine therapy. Key words: Shuganyishen formula; Hormone-dependant metastatic breast cancer

Selective Effects of Glucose, Insulin and Leptin by Molecular Breast Cancer Subtype.

Abstract Background: Type II diabetes and obesity are important risk factors for post-menopausal luminal A (LA) and triple negative (TN) pre-menopausal breast cancers, particularly in African American (AA) and Hispanic women. Breast cancer patients with these chronic metabolic diseases also have a worse prognosis, independent of other factors. We have published that metformin inhibits cell growth (S phase arrest) and induces apoptosis, only in TN cell lines in vivo and in vitro. It is less active, growth inhibitory (G1 arrest) and does not induce apoptosis in other breast cancer cell subtypes (Cell Cycle, 2009).Methods: We investigated the effects of glucose as a mitogen at physiologic (5mM), metabolic syndrome (7mM) or diabetic levels (10mM), with or without insulin (100 ng/ml) or leptin (100 ng/ml) using cell lines representing all molecular subtypes of breast cancer. Metformin was then used in combination with the above, to determine whether it would block the mitogenic or signaling effects of supraphysiological glucose, insulin or leptin.Results: The LA (MCF-7) and 2 of 5 TN cell lines (derived from AA patients; HCC 1806 and MDA 468) showed the most cell growth in response to glucose &amp;gt;5mM (75% for MCF-7, 30-50% for AA TN lines). Three TN cell lines from Caucasians (MDA231, HCC1937, BT20), the HER2 (SKBR3) and luminal B (BT-474) cells showed less growth induction with glucose &amp;gt;5mM. In TN lines only, glucose associated mitogenesis was associated with increased EGFR, pEGFR, IGF1R, pIGF1R, AKT and pAKT and decreases in AMPK, pAMPK, p38, IRS2, and the cyclins D1, E and A in a dose dependent manner. Metformin abrogated glucose induced cell growth and the aforementioned protein expression/phosphorylation changes involving EGFR, IGF1R, and AKT, increased AMPK and pAMPK and induced a profound reduction in Cyclin D1 across all glucose concentrations in TN cell lines from AA women. It reduced but did not eliminate glucose associated mitogenesis in the TN cell lines from Caucasian patients. Metformin had a more variable effect on cell lines of other molecular subtypes grown under high glucose conditions. The 5 TN breast cancer cell lines were uniformly resistant to both leptin and insulin associated mitogenesis, across a wide range of glucose concentrations. In contrast, both leptin and insulin significantly promoted LA breast cancer cell growth. These effects were resistant to metformin treatment. Leptin and insulin had the least growth promoting effects on HER2 breast cancer cell lines, whereas they induced modest growth induction in LB cell lines.Conclusions: All TN cell lines showed significant mitogenesis in response to glucose &amp;gt;5mM, whereas they were uniformly resistant to both leptin and insulin. The glucose associated mitogenesis was more pronounced in lines derived from AA patients, as were the anti-mitgenic effects of metformin. LA cells showed marked growth induction by glucose, leptin and insulin, whereas HER2 cell lines showed general resistance to all of these factors. These data suggest that metabolic and hormonal shifts with obesity and diabetes, as well as metformin response vary by the molecular subtype of breast cancer cells and ethnicity. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 5154.