Breast Caner Research Articles

ZNF775 inhibits MCF-7 breast cancer cell migration by downregulating Wnt5a

C2H2 zinc finger protein is widely involved in the occurrence and development of cancer. However, the function and mechanism of most C2H2 zinc finger proteins in breast caner (BC) remains unclear. Here, we reported the expression prognosis of C2H2 type zinc finger protein ZNF775 in BC patients and its possible biological mechanism. First, multiple public databases showed that ZNF775 was significantly overexpressed in BC tissues. Interestingly, high expression of ZNF775 was significantly associated with a better prognosis. Immunohistochemistry were used for verification, and the expression of ZNF775 was consistent with the databases. Considering the large heterogeneity of different breast cancer cells, we temporarily selected MCF-7 cell line for verification. In vitro overexpression experiments showed that overexpression of ZNF775 significantly inhibited the proliferation and migration of MCF-7 BC cell. We further combined RNA-sequencing (RNA-seq) and CUT & Tag, and found that overexpression of ZNF775 can down-regulate the expression of most genes in the Wnt signaling pathway. The cBioportal database showed that ZNF775 was negatively correlated with the expression of Wnt5a, suggesting that its downstream target was likely Wnt5a. Finally, we discovered that Wnt5a could partially reverse the inhibitory effect of ZNF775 on MCF-7 BC cell migration through transwell migration experiments. In conclusion, our findings will provide new ideas for the diagnosis, treatment and prognosis assessment of BC in the future.

Analisis Perbandingan Pengukuran Jarak Algoritma K-Nearest Neighbor Dengan Menggunakan Data Breast Cancer Dan Data Heart Disease


 
 
 
 Breast Cancer is a cancerous condition that appears in the breast area. This type of cancer is often experienced by women with a characteristic feature of Breast Cancer, namely the appearance of unusual lumps in the breast area. Heart or Heart Disease is a type of Non-Communicable Disease (PTM): which results in a fairly high mortality rate. Heart Disease is caused by several risk factors including smoking, an unhealthy lifestyle, high cholesterol, hypertension, and diabetes.
 Based on these facts, an appropriate algorithm is needed to classify Breast Caner and Heart Disease as an effort to prevent an increase in mortality rates due to Breast Cancer and Heart Disease. And the algorithm that will be used is the K-Nearest Neighbor algorithm with 3 distance measurement methods, namely Euclidean distance, Manhattan distance, and Minkowsky distance .
 
 
 
 
 From the stages that have been carried out, the final results of the Euclidean distance method obtained an Accuracy value of 80.88% Breast Cancer data at K = 11, and 78.69% heart Disease data at K = 11. The Manhattan distance method obtained an Accuracy value of 89.71% of Breast Cancer data on K=11, and 78.69% of Heart Disease data on K=20.The Minkowsky distance method obtained an Accuracy value of 98.53% of Breast Cancer data on K=11, and 79.41% of Heart Disease data on K=11. This shows that the Minkowsky distance method works more optimally than the Euclidean distance and Manhattan distance methods.

Evaluation of sister chromatid exchange in patients with breast cancer in relation to clinical stage

High levels of sister chromtid exchanges (SCE) in patients with breast caner reflects a genomic instability that may be operative in carcinogenesis.The frequency of sister chromatid exchanges in peripheral blood lymphocytes was analyzed in patients with breast cancer in relation to clinical stage in 12 stage II and 5 Stage IV untreated breast cancer patients.Moreover,SCE baseline values in patients were compared with a control group of 12 healthy women. A significant difference emerged between patients and control (P<0.01).SCE increased significantly in stage IV breast cancer patients compared with those patients of stage II (P<0.05).The results reveal that SCE frequency increases with the progression of clinical stage of breast cancer.

Prognostic factors and survival outcomes according to tumor subtype in patients with breast cancer lung metastases.

BackgroundReports on the incidence and prognoses of lung metastases when diagnosing breast cancer patients with different subtypes are limited. Our study investigated the effect of molecular sub-typing stratification on the prognoses of lung metastatic breast caner patients.MethodsPatients with breast cancer and lung metastases were identified from Surveillance, Epidemiology and End Results population-based data between 2010 and 2015. Univariate and multivariate Cox regression analyses were performed to identify risk factors and prognoses, overall survival (OS) and breast cancer-specific survival for patients with breast cancer lung metastases.ResultsWe identified 6,516 patients with lung metastatic breast cancer, representing 1.7% of the entire cohort and 30.4% of the subset with metastatic disease. This included 2,940 hormone receptor (HR)+/HER2− patients, 852 HR+/HER2+ patients, 547 HR−/HER2+ patients and 983 triple-negative patients. The median OS for all lung metastatic patients was 13 months. Multivariate analysis revealed that those lung metastatic breast cancer patients of older age (>80), black race, with poorly differentiated tumors, carcinoma histology, triple-negative subtype, more metastatic sites and no surgery, and no chemotherapy showed significantly poor survival, both overall and breast cancer-specific.ConclusionsOur findings show that molecular sub-type and more metastatic sites might have significant influence on the incidence and prognosis of breast cancer lung metastases. We also identified several prognostic factors that could guide therapy selection in the treatment of lung metastatic patients.

9PD - Aurora-B mediated snail phosphorylation is essential for mitotic spindle checkpoint and for preventing chromosomal instability in breast cancer

BackgroundSnail is a transcriptional repressor that promotes epithelial-to-mesenchymal transition (EMT). Snail is upregulated in breast cancer and it is associated with chromosomal instability (CIN) and metastasis. The mitotic spindle assembly checkpoint (SAC) is a critical mechanism to prevent CIN. Extensive studies have investigated signaling pathways that involve in protein-protein interactions and post translational modifications. However, it not known how mitosis might be impacted by transcriptional regulation. Due to the fact that Snail is a transcriptional regulator and that Snail is upregulated in breast cancer with CIN, it is critical to understand whether and how abnormally regulated Snail contributes to breast cancer CIN via transcriptional regulation. MethodsWe used siRNA to knockdown Snail in breast caner cell lines to look for a change in SAC. We developed a phosphor-specific antibody to study Aurora-B mediated phosphorylation. Site specific mutagenesis was used to generate phosphorylation site mutants, and exogenous proteins were expressed in cells to look for an effect in SAC and CIN. ResultsWe found that Snail is an essential element that is required for the SAC. Genetic depletion of Snail in breast cancer cells by siRNA led to a defect in activation of SAC (measured by accumulation of Histone Serine 10 phosphorylation in response to spindle damaging agents). Interestingly we also found that the Snail protein level was significantly enhanced in the presence of nocodazole. Further, we found that Snail upregulation was through phosphorylation in an Aurora-B dependent manner. We identified a serine site that can be phosphorylated by Aurora-B. Using a phosphor-specific antibody we have developed, we further proved that the phosphorylation event was initiated in mitosis. Functional studies reveal a critical role for Snail phosphorylation in activation of the SAC and prevention of CIN. Conclusions1. Snail is required for mitotic spindle checkpoint activation; 2. Snail is phosphorylated by Aurora B in mitosis; 3. Aurora-B medicated Snail phosphorylation is required for optinal activation of the spindle checkpoint and prevention of chromosomal instability in breast cancer. Legal entity responsible for the studyThe authors. FundingTianjin Medical University Cancer Institute and Hospital. DisclosureAll authors have declared no conflicts of interest.

Long non-coding RNA-FOXF1 adjacent non-coding RNA as a good prognostic factor in breast cancer

Objective To test FOXF1 adjacent non-coding RNA (FENDRR) expression levels in breast cancer samples, and evaluate its effects on recurrence-free survival (RFS) in breast cancer and how FENDRR modulates drug sensitivity to doxorubicin. Methods Real-time quantitative polymerase chain reaction (Real-time PCR) and pared t test were utilized to analyze FENDRR levels in normal breast tissues and cancer tissues. Online survival analysis website was used to test the effects of FENDRR on breast cancer survival. Small interfering RNAs (siRNAs) were used to knock down FENDRR and SRB was carried out to test the response to doxorubicin. We also tested the mRNA levels of adenosine triphosphate binding cassette subfamily B member 1 (ABCB1) and multidrug resistance associated protein 1 (ABCC1) after down-regulating FENDRR. Results The FENDRR levels were lower in cancer samples than those in normal breast tissues in breast caner (1.00±0.13 vs. 2.50±0.34, t=4.528, P<0.01). RFS rate of breast cancer patients with lower levels of FENDRR was lower in all patients, the subgroups of luminal A, luminal B and basal, and patients receiving systemic treatment [P<0.01, <0.01, <0.05, <0.05 and <0.01, hazard ratio (HR)=0.67 (0.57-0.79), 0.60 (0.47-0.78), 0.70 (0.51-0.96), 0.65 (0.47-0.91) and 0.61 (0.49-0.76)]. Down-regulation of FENDRR could up-regulate the expression of ABCB1 and ABCC1, conferring resistance to doxorubicin in MCF-7 cells. Conclusion FENDRR was lower in breast cancer samples, and correlated to better survival in breast cancer. By regulating MDR-related gene expression, FENDRR modulated drug sensitivity in MCF-7 cells. Key words: Breast cancer; Long non-coding RNA; Survival; Drug resistance

TS-1 add-on therapy in Japanese patients with triple-negative breast cancer after neoadjuvant or adjuvant chemotherapy: a feasibility study

SummaryPurpose We examined the feasibility, efficacy, and safety of TS-1 add-on therapy (TAT) in Japanese patients with triple-negative breast caner (TNBC). Methods TAT (TS-1, 80 mg/m2/day, BID, PO), consisting of the 21-day cycles of 14-day consecutive administration followed by 7-day drug holiday, was conducted for 365 days. The median follow-up was 75.2 months (range, 7.3–103.3 months). The primary endpoint was the feasibility of TAT. The secondary endpoints included relapse-free survival (RFS), overall survival (OS), and safety. Results 63 Japanese patients with TNBC (median age, 52.5 years; range, 23.7–68.6 years) were examined. Among them, 34 (54.0%) were postmenopausal, 54 (93.7%) had TNBC of common histological type, 51 (81.0%) had T1 to 3 tumors, 63 (100%) had undergone standardized surgery, and 44 (69.8%) and 19 (30.2%) had undergone neoadjuvant chemotherapy and adjuvant chemotherapy, respectively. The 365-day cumulative rate of TS-1 administration was 68.3% (95% confidence interval, 55.3–79.4), being comparable to 65.8% previously reported for gastric cancer. The 5-year RFS rates were 52.3% and 84.2% in the neoadjuvant and adjuvant chemotherapy groups, respectively, and the 5-year OS rates were 68.0% and 89.5%, respectively. The most common adverse events (AEs) were leucocyte count decreased (50.8%), total bilirubin decreased (44.4%), and pigmentation (42.9%). AEs were manageable clinically, and any grade 4 AEs did not develop. Conclusions The 365-day cumulative rate of TS-1 administration in TNBC patients was comparable to that in gastric cancer patients despite previous chemotherapy with anthracyclines and/or taxanes. TAT was feasible for TNBC patients after standard primary therapy.

Prognostic value of Tfh-like cells in breast cancer.

e14285 Background: The treatment of breast cancer, one of the most common malignant tumors for female, focuses on the combined treatments based on subtypes including Luminal A, Luminal B, HER2 overexpression and triple negative. Given that none of those subtypes takes into consideration of immunological parameters, it is valuable to explore the biological characteristics and prognostic value of immune cells in breast caner. T follicular helper cells (Tfh cells) have been identified in different types of tumors with rare datas in breast cancer. One of them indicated the positive prognostic value of 8-gene Tfh signature. Exploration of Tfh cells may provide new clues to the potential immunotherapies in breast cancer. Methods: Freshly resected invasive breast cancer tissue from Fudan University Shanghai Cancer Center were collected during 06/2015 to 05/2017. Patients were divided into Luminal A (N = 61), Luminal B (HER2-) (N = 138), Luminal B (HER2+) (N = 72), HER2+ (non-luminal) (N = 57) and triple negative (N = 58) based on St Gallen International Expert Consensus. Results: A subpopulation of PD1hiCD4+ T cells in the tumor tissues of breast cancer was identified as Tfh-like cells, with the specific expression of Bcl-6 and CXCL13, confirmed by IHC. Phenotypes of those cells were checked by flow cytometry. Tfh-like cells were of a high level of ICOS but negative for CXCR5, suggesting that they were atypical Tfh cells. Data showed that high grade (N = 166; mean±SEM, 17.62±0.87) compared with low grade (N = 220; mean±SEM, 12.41±0.46) or high Ki-67 level (N = 300; mean±SEM, 15.38±0.58) with low Ki-67 level (N = 86; mean±SEM, 12.10±0.67), and negative ER expression (N = 115; mean±SEM, 17.47±1.05) compared with positive ER expression (N = 271; mean±SEM, 13.46±0.50) or negative PR expression (N = 177; mean±SEM, 16.87±0.83) compared with positive PR expression (N = 209; mean±SEM, 12.77±0.50) was associated with higher frequency of Tfh-like cells (P &lt; 0.01 to all). Patients with triple negative had higher frequency of Tfh-like cells than those with Luminal A (difference, 6.83; P = 0.0006) and Luminal B (HER2-) (difference, 4.61; P = 0.0124). Patients with HER2+ (non-luminal) had higher frequency of Tfh-like cells than those with Luminal A (difference, 5.34; P = 0.0146). These results indicate that higher frequency of Tfh-like cells could have negative prognostic influence. Conclusions: Our study defined a PD1hiBcl6+CXCL13+CD4+ T cell subpopulation, which specifically secretes cytokine IL-21 as Tfh-like cells. Higher frequency of Tfh-like cells is more likely to be seen in patients with poor prognosis.

An 8 Years Retrospective Study of Breast Cancer Incidence in Ilam Province, Western Iran

Breast cancer is the most common cancer (27% of all cancers) and common cause of death (16%) which occurs due to cancers among women, either in developed or developing countries. The current study aimed to assess incidence of breast cancer in west of Iran (Ilam province). During a cross-sectional study, all documented records of patients who were referred to the health centre of Ilam province in a period of 8 years (2002-2009) were investigated and 82 files which were related to breast cancer were identified. Patients' data were entered into SPSS, version 16 and using X2, t-test and descriptive statistics, they were analyzed. Totally, 82 confirmed breast cancer cases were diagnosed between 2002-2009 and this figure accounted for 21.4% of all cancers in Ilam province. The mean age with standard deviation (SD) of patients was 47.4 ± 14.5 years and the disease was most frequent in the age group of 30-40 years (38.3%). The highest incidence rate was seen in 2006- 2007 and the lowest rate was seen in 2002-2003. The most prevalent morphologic pattern of breast caner (86.2%) was invasive ductal carcinoma (IDC). There was a 23% incidence rate for breast cancer, with a significant increase in its incidence rate during 2002-2009. Due to diagnosis of the disease in its advanced stages, and also involvement of low age groups and young population in the country, screening programs such as self examination, examination by doctors and mammography should be started in the lower age groups, in ages which are lower than 30 years.

Anti-HER-2 therapy following severe trastuzumab-induced cardiac toxicity

The human epidermal growth factor receptor 2 (HER2) is overexpressed in 25%–30% of breast cancer patients. Anti-HER2 therapies have changed the aggressive course of HER2+ breast cancer. In spite of the therapeutic benefits, their cardiotoxicities are major concerns, especially when used concurrently with anthracyclines.Here we present an elderly patient with relapsed HER2+ breast cancer. Her presentation for relapsed disease was unusual for the physical finding as well as the history of trastuzumab-induced severe cardiotoxicity while requiring additional anti-HER2 therapy. She received neoadjuvant anti-HER2 treatment for stage III breast caner. Due to severe reduction of cardiac ejection fraction (EF), she only received five doses of adjuvant transtuzumab. Unfortunately her disease relapsed one year later with chest wall lesions and a persistent low EF. We treated the patient with lapatinib combined with capecitabine which resulted rapid resolution of her chest wall lesion. More importantly, the patient had one year of disease control without deterioration in her ejection fraction. We discussed the management of recurrent HER2+ breast cancer with chest wall disease and the choice of anti-HER2 therapy in patients with a history of transtuzumab-induced cardiac dysfunction.

TRPV4 plays a role in breast cancer cell migration via Ca2+-dependent activation of AKT and downregulation of E-cadherin cell cortex protein

TRPV4 belongs to the ‘Transient Receptor Potential’ (TRP) superfamily. It has been identified to profoundly affect a variety of physiological processes, including nociception, heat sensation and inflammation. Unlike other TRP superfamily channels, its role in cancers are unknown until recently when we reported TRPV4 to be required for cancer cell softness that may promote breast cancer cell extravasation and metastasis. Here, we elucidated the molecular mechanisms mediated by TRPV4 in the metastatic breast cancer cells. TRPV4-mediated signaling was demonstrated to involve Ca2+-dependent activation of AKT and downregulation of E-cadherin expression, which was abolished upon TRPV4 silencing. Functionally, TRPV4-enhanced breast caner cell transendothelial migration requires AKT activity while a combination of transcriptional and post-translational regulation contributed to the TRPV4-mediated E-cadherin downregulation. Finally, mass spectrometry analysis revealed that TRPV4 is required for the expression of a network of secreted proteins involved in extracellular matrix remodeling. In conclusion, TRPV4 may regulate breast cancer metastasis by regulating cell softness through the Ca2+-dependent AKT-E-cadherin signaling axis and regulation of the expression of extracellular proteins.

Effect of long noncoding RNA PANDAR on the proliferation and invasion of breast cancer cells

Objective To investigate the effect of long noncoding RNA (lncRNA) PANDAR on the proliferation and invasion of breast cancer cells. Methods We constructed the stable MCF-7 cells with overexpression or knockdown of PANDAR by using lentivirus system and detected the cell proliferation and invasion by cell counting kit-8 (CCK-8) assay and matrigel Transwell assay. Results The relative expression of PANDAR in MCF-7 cells with overexpression of PANDAR was 20.05±1.08, higher than in the control group (P=0.000), and that in MCF-7 cells with knockdown of PANDAR was 0.16±0.02, lower than in the control group (P=0.008). The cell proliferation activity of PANDAR overexpression group (0.489±0.067, 0.945±0.025, 1.236±0.072 ) at 24, 48 and 72 h, were higher than these in control group (0.385±0.024, 0.576±0.059, 0.737±0.041; P=0.045, P=0.024, P=0.012, respectively). The cell proliferation activity of PANDAR knockdown group (0.292±0.055, 0.371±0.041, 0.455±0.068) at 24, 48 and 72 h, were significantly lower than these in control group (P=0.036, P=0.030, P=0.014), respectively. The amount of invasion cells in PANDAR overexpression group were significantly more than control group (48.37±13.08 vs. 25.15±6.75; P=0. 011). While the amount of invasion cells in PANDAR knockdown group were significantly less than control group (23.35±2.96 vs. 50.13±11.87; P=0. 015). Conclusion PANDAR promotes cell proliferation and invasion of breast caner cells. Key words: Long noncoding RNA; Breast cancer

Neoadjuvant Chemotherapy Creates Surgery Opportunities For Inoperable Locally Advanced Breast Cancer

Neoadjuvant chemotherapy (NAC), the systematic chemotherapy given to patients with locally advanced and inoperable breast caner, has been proven to be of great clinical values. Many scientific reports confirmed NAC could effectively eliminate sub-clinical disseminated lesions of tumor, and improve long-term and disease-free survival rate of patients with locally advanced breast cancer (LABC); however, up to now, LABC is still a serious clinical issue given improved screening and early diagnosis. This study, with main focus on inoperable LABC, investigated the values of NAC in converting inoperable LABC into operable status and assessed the prognosis. Sixty-one patients with inoperable LABC were initially treated with neoadjuvant chemotherapy; their local conditions were improved to operable status. Radical surgery was exerted on 49 patients. Original chemotherapy was performed after surgery, followed by local radiotherapy. And endocrine therapy was optional according to the hormone receptor status. The quality of life for most patients with skin diabrosis was obviously improved because their local conditions were under control. For all recruited cases, the survival duration and life quality were significantly improved in patients who finished both NAC and surgery compared to those who did not. Further more, this study demonstrates improved prognostic consequences.

Correlation of AS160 hyperphosphorylation and 18F-FDG uptaking in invasive breast cancer

Objective To investigate the correlation between AS160 hyperphosphorylation and 18F-fluorodeoxyglucose (18F-FDG) uptaking in invasive breast cancer. Methods Thirty-nine primary breast caner patients received PET/CT scan before operation and their maximum standard uptake values (SUVmax) were recorded. The levels of p-AS160 (Thr642) were detected using immunohistochemical method in breast cancer tissues. The association between p-AS160 (Thr642) levels and SUVmax was assessed using the Spearman nonparametric test. Results The 18F-FDG uptake of tumors of the 39 patients with primary breast caner was 6.1±2.7 (SUVmax). There was one false negative case in PET/CT scan imaging, compared with postoperative pathologic result. The positive rate of p-AS160 (Thr642) was 76.9%, and it was positively correlated with SUVmax (r=0.672, P<0.001). Conclusion AS160 plays a significant role in the glucose metablism of breast cancer tissues and its hyperphosphorylation may contribute to the increased uptaking of glucose in patients with breast cancer. Key words: Breast neoplasms; Phosphorylation; Fluomdeoxyglucose F18; AS160

Abstract 3325: Combination of DNMT and HDAC inhibitors reprogram cancer stem cell signaling to overcome drug resistance

Abstract In recent years, impressive technical advancements have been made in the isolation and validation of the mammary stem cells (MaSCs) and cancer stem cells (CSCs). However, the signaling pathways that regulate stem cell self-renewal are largely unknown. Further, CSCs are believed to contribute to resistance to chemotherapy and radiation therapy. However, an effective therapeutic strategy to overcome this resistance is yet to be identified. We have recently discovered that the DNA methyltransferases, especially DNMT1, play critical role in MaSCs and CSCs self-renewal and targeted deletion of this gene impaired mammary tumor formation by inhibiting CSCs formation. However, the molecular mechanism(s) by which DNMTs control CSCs and the therapeutic relevance of DNMTs inhibitors in regulation of CSCs and overcome drug resistance are also largely unknown. In this study, using MMTV-Neu-Tg mouse mammary tumor model, we found that both luminal progenitor and basal stem cells are susceptible to genetic and epigenetic modifications, which leads to activation of un-activated Neu-Tg into transformed tumor forming phenotype. Combination of 5-Azacytidine, a DNMT inhibitor, and butyrate, a HDAC inhibitor, markedly reduces CSCs and consequently increases the overall survival of the animal. RNA-seq analysis of the CSCs treated with 5-AzaC+butyrate provides evidence that combined inhibition of DNMTs and HDACs reduces CSCs pool in the mammary gland by blocking growth-promoting signaling molecules like RAD51AP1 and SPC25. RAD51AP1 and SPC25, which are known to play a key role in DNA damage repair and kinetochore assembly, are significantly overexpressed in breast tumor tissues and associated with decreased overall patients’ survival. Further, these two genes are overexpressed in Tamoxifen and Taxol resistant human breast cancer cell lines. Functional inactivation of these genes in breast caner cells facilitates chemotherapy-induced apoptosis and reduces tumor growth. Overall, our studies provide strong evidence that breast CSCs (both basal stem cells and luminal progenitor cells) are susceptible for genetic and epigenetic modifications and associated with resistance to chemo- and radiotherapy. Thus, combination of DNMT and HDAC inhibitors can serve as an effective therapeutic strategy to block mammary tumor growth and to overcome drug resistance by inhibiting CSCs. Citation Format: Rajneesh Pathania, Ravindra B. Kolhe, Sabarish Ramachandran, Gurusamy Mariappan, Priyanka Thakur, Puttur D. Prasad, Vadivel Ganapathy, Muthusamy Thangaraju. Combination of DNMT and HDAC inhibitors reprogram cancer stem cell signaling to overcome drug resistance. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3325.

Dihydroartemisinin prevents breast cancer-induced osteolysis via inhibiting both breast caner cells and osteoclasts.

Bone is the most common site of distant relapse in breast cancer, leading to severe complications which dramatically affect the patients’ quality of life. It is believed that the crosstalk between metastatic breast cancer cells and osteoclasts is critical for breast cancer-induced osteolysis. In this study, the effects of dihydroartemisinin (DHA) on osteoclast formation, bone resorption, osteoblast differentiation and mineralization were initially assessed in vitro, followed by further investigation in a titanium-particle-induced osteolysis model in vivo. Based on the proved inhibitory effect of DHA on osteolysis, DHA was further applied to MDA-MB-231 breast cancer-induced mouse osteolysis model, with the underlying molecular mechanisms further investigated. Here, we verified for the first time that DHA suppressed osteoclast differentiation, F-actin ring formation and bone resorption through suppressing AKT/SRC pathways, leading to the preventive effect of DHA on titanium-particle-induced osteolysis without affecting osteoblast function. More importantly, we demonstrated that DHA inhibited breast tumor-induced osteolysis through inhibiting the proliferation, migration and invasion of MDA-MB-231 cells via modulating AKT signaling pathway. In conclusion, DHA effectively inhibited osteoclastogenesis and prevented breast cancer-induced osteolysis.

The expression and clinical significance of CD47 and calreticulin in infiltrating ductal breast carcinomas

Objective To evaluate the expression of CD47 and calreticulin and their relationships with clinicopathologic features, prognosis in infiltrating ductal breast carcinomas. Methods The expression of CD47 and calreticulin was evaluated in 128 cases with infiltrating ductal breast carcinomas by means of immunohistochemistry, and the relationship between CD47 and other clinicopathologic factors as well as prognosis were analyzed. Results The expression level of CD47 in cancer nest was 81/128, which was lower than 106/128 in its adjacent normal epithelium (χ2=12.400,P<0.05),but calreticulin expression was 50/128 and was higher in cancer nest(χ2=21.510,P< 0.05).In addition,the expression level of calreticulin had certain difference in different molecular subtype(χ2= 21.510,P<0.05),and an increased expression in Basal-like and Her-(2+)/ER(-)subtypes.The expression level of CD47 was associated with tumor sizes,histological tumor grade and lymph node involvement(χ2=11.400,4.732,5.432,all P<0.05),and the expression level of calreticulin was associated with histological tumor grade and distant metastasis(χ2=10.810,6.770,all P<0.05).The 5-year survival rate of patients performed following-up was 68.37%. Univariate analysis indicated the survival rate in 5 years of high level CD47,calreticulin expression was higher than that of low level CD47,calreticulin expression(χ2=5.231,13.069,all P<0.05). Conclusion The expression of CD47, calreticulin is associated with index of dysprognosis and survival rate in 5 years, so combined CD47 and calreticulin detect may be of prognostic value in patients with breast caner. Key words: CD47; Calreticulin; Breast neoplasms; Immunohistochemistry

Abstract 4421: GPER-mediated activation of the cAMP response element-binding protein (CREB) and subsequent IGFBP-1 expression is altered during the development of tamoxifen resistance

Abstract Tamoxifen is a commonly prescribed anti-estrogen treatment for breast cancer patients; this selective estrogen receptor modulator functions as an ERα-antagonist in breast tissue and inhibits cell proliferation and survival of breast cancer cells. More recently, the active metabolite of tamoxifen, 4-OHT (Tam), has been shown to activate the G protein-coupled estrogen receptor (GPER). Previous studies from our laboratory identified a novel, GPER-dependent mechanism of Tam action in breast cancer cells that requires the extracellular accumulation of the insulin-like growth factor binding protein-1 (IGFBP-1). The molecular mechanisms of Tam-induced IGFBP-1 production mediated by GPER have not been determined. GPER activation has been shown to stimulate cAMP production and PKA activation in several cell types. We hypothesized the cAMP response element-binding protein (CREB) activation is required for Tam-induced IGFBP-1 production in breast caner cells. To determine if CREB activation may be involved in IGFBP-1 transcription, CREB phosphorylation (S133) status and nuclear accumulation were determined in MCF-7 cells after Tam treatment. The phosphorylation and nuclear accumulation of CREB were observed in Tam-treated MCF-7 cells suggesting that CREB activation may be required for Tam-induced IGFBP-1 transcription. Increased CREB phosphorylation and IGFBP-1 transcription were inhibited by pretreatment with the PKA inhibitor H-89 supporting a role for CREB in Tam-induced IGFBP-1 expression. H-89 pretreatment also significantly decreased the previously reported inhibition of IGF-1-stimulated signaling by Tam treatment in breast cancer cells. We have previously reported that Tam-induced IGFBP-1 expression is dysregulated in Tam-resistant MCF-7 cells (MCF-7TamR). To determine if aberrant CREB signaling results in the observed dysregulation, similar experiments were performed in MCF-7TamR cells. Our results show that CREB phosphorylation and nuclear accumulation are decreased in Tam-resistant cells compared to Tam-sensitive cells. Taken together, these results suggest that activation of GPER results in CREB-dependent IGFBP-1 production and subsequent inhibition of IGF-1-stimulated signaling in breast cancer cells treated with Tam. Furthermore, dysregulation of GPER-mediated CREB activation may be involved in the development of Tam resistance in breast cancer. Citation Format: Ali Vaziri-Gohar, Kevin D. Houston. GPER-mediated activation of the cAMP response element-binding protein (CREB) and subsequent IGFBP-1 expression is altered during the development of tamoxifen resistance. [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 4421. doi:10.1158/1538-7445.AM2015-4421

Efficacy and Safety of HER2-Targeted Agents for Breast Cancer with HER2-Overexpression: A Network Meta-Analysis.

BackgroundClinical trials of human epidermal growth factor receptor 2 (HER2)-targeted agents added to standard treatment have been efficacious for HER2-positive (HER2+) advanced breast cancer. To our knowledge, no meta-analysis has evaluated HER2-targeted therapy including trastuzumab emtansine (T-DM1) and pertuzumab for HER2-positive breast caner and ranked the targeted treatments. We performed a network meta-analysis of both direct and indirect comparisons to evaluate the effect of adding HER2-targeted agents to standard treatment and examined side effects.MethodsWe performed a Bayesian-framework network meta-analysis of randomized controlled trials to compare 6 HER2-targeted treatment regimens and 1 naïve standard treatment (NST, without any-targeted drugs) in targeted treatment of HER2+ breast cancer in adults. These treatment regimens were T-DM1, LC (lapatinib), HC (trastuzumab), PEC (pertuzumab), LHC (lapatinib and trastuzumab), and PEHC (pertuzumab and trastuzumab). The main outcomes were overall survival and response rates. We also examined side effects of rash, LVEF (left ventricular ejection fraction), fatigue, and gastrointestinal disorders, and performed subgroup analysis for the different treatment regimens in metastatic or advanced breast cancer.ResultsWe identified 25 articles of 21 trials, with data for 11,276 participants. T-DM1 and PEHC were more efficient drug regimens with regard to overall survival as compared with LHC, LC, HC and PEC. The incidence of treatment-related rash occurs more frequently in the patients who received LC treatment regimen than PEHC and T-DM1 and HC. In subgroup analysis, T-DM1 was associated with increased overall survival as compared with LC and HC. PEHC was associated with increased overall response as compared with LC, HC, and NST.ConclusionsOverall, the regimen of T-DM1 as well as pertuzumab in combination with trastuzumab and docetaxel is efficacious with fewer side effects as compared with other regimens, especially for advanced HER2+ breast cancer.ImpactThis study suggests that both T-DM1 and PEHC therapy are potentially and equally useful treatments for HER2+ breast cancer.

MiR-491-5p functions as a tumor suppressor by targeting JMJD2B in ERα-positive breast cancer

The involvement of miR-491-5p in breast cancer development is unclear. This study showed that miR-491-5p is significantly downregulated in ERα-positive breast cancer tissues and cell lines and is generally hypermethylated in ERα-positive breast cancer. MiR-491-5p overexpression significantly suppressed estrogen signaling and estrogen-stimulated proliferation of breast cancer cells. Furthermore, the histone demethylase JMJD2B was identified as a direct target of miR-491-5p. The ectopic expression of JMJD2B abrogated the phenotypic changes induced by miR-491-5p in breast cancer cells. Collectively, our data indicate that miR-491-5p plays a tumor suppressor role in the development and progression of breast caner and may be a novel therapeutic target against ERα-positive breast cancer.