Arctigenin (Arc), a novel anti-inflammatory lignan derived primarily from Arctium lappa, has demonstrated promising anticancer activity in multiple cancer types. This study was designed to evaluate the anticancer efficacy of Arc across distinct molecular subtypes of breast cancer in vitro and in vivo and to gain mechanistic insights into its mode of action. In vitro evaluation was conducted in estrogen-receptor-positive MCF-7, human epidermal growth factor receptor 2 (HER2)-positive SKBR3, and triple-negative MDA-MB-231 breast cancer cell lines. In vivo efficacy and safety were evaluated using female severe combined immunodeficient (SCID) mice (5-7 weeks old) bearing MCF-7 or MDA-MB-231 xenografts. Mice received daily oral gavage of Arc at 50 mg/kg body weight for 8 weeks. In vitro, Arc inhibited cell proliferation across all three breast cancer subtypes in a dose-dependent manner. PCR-array analysis of gene expression revealed that Arc targets multiple signaling molecules involved in cell proliferation, cell cycle regulation, apoptosis, migration/invasion, and drug transport, demonstrating a subtype-specific target profile. Arc induced cell-cycle arrest at the G2/M phase in MCF-7 cells and at G0/G1 in MDA-MB-231 cells, accompanied by significant induction of apoptosis in both cell lines. Migration assays further demonstrated marked inhibition of wound closure in Arc-treated cells. In vivo, Arc treatment significantly inhibited tumor growth in both xenograft models, decreased Ki67 expression, and produced no overt toxicity. In summary, Arc exhibits potent anticancer activity against distinct breast cancer subtypes through multi-targeting mechanisms. Given the heterogeneity of breast cancer, Arc appears to be a promising candidate for further preclinical investigation.

Background: Obesity is associated with aggressive prostate cancer, but the links between metabolic dysregulation, inflammation, adipocyte-associated signaling, and tumor growth remain incompletely defined. This study examined whether high-fat diet (HFD)-associated systemic changes and adipocyte-derived paracrine interactions are linked to prostate cancer growth in preclinical models. Methods: An HFD xenograft model and adipocyte co-culture systems were used to evaluate systemic and local tumor-supportive features. Pharmacologic/nutritional modulation was tested using green tea or EGCG, arctigenin, and the CCR2 antagonist RS 504393, alone or in combination. Tumor growth, cell proliferation, angiogenesis-related features, circulating metabolic and cytokine levels, and selected tumor-associated signaling proteins were analyzed. Results: HFD feeding was associated with increased circulating free fatty acids, IGF-1, MCP-1, IL-6, and VEGF, together with increased tumor growth, Ki67 staining, and CD31-positive microvessel density. Adipocyte co-culture systems were used to evaluate treatment-associated changes in prostate cancer cell proliferation under adipocyte-associated conditions. Combined modulation with green tea/EGCG, arctigenin, and RS 504393 was associated with greater reductions in adipocyte-associated proliferation, tumor growth, Ki67 staining, and CD31-positive microvessel density than single or dual interventions. Antibody array analysis showed treatment-associated changes in selected stress- and apoptosis-related proteins, including cleaved caspase-7 and phosphorylated Chk1. Conclusions: HFD-associated metabolic and inflammatory alterations, adipocyte-associated interactions, proliferative activity, angiogenesis-related features, and stress/apoptosis-related signaling changes were linked within a tumor-supportive framework in preclinical prostate cancer models. Combined pharmacologic/nutritional modulation was associated with reduced tumor-supportive features under HFD conditions. Further mechanistic and translational validation is needed.

Breast cancer (BC) remains among the most commonly diagnosed cancers in women worldwide. Triple-negative BC (TNBC) is a subset of BC characterized by aggressive behavior, a high risk of distant recurrence, and poor overall survival rates. Chemotherapy is the backbone for treatment in patients with TNBC, but outcomes remain poor compared to other BC subtypes, in part due to the lack of recognized functional targets. In this study, the expression of the tetraspan protein epithelial membrane protein 2 (EMP2) was explored as a predictor of TNBC response to standard chemotherapy. We demonstrate that EMP2 functions as a prognostic biomarker for patients treated with taxane-based chemotherapy, with high expression at both transcriptomic and protein levels following treatment correlating with poor overall survival. Moreover, we show that targeting EMP2 in combination with docetaxel reduces tumor load in syngeneic and xenograft models of TNBC. These results provide support for the prognostic and therapeutic potential of this tetraspan protein, suggesting that anti-EMP2 therapy may be beneficial for the treatment of select chemotherapy-resistant TNBC tumors.

Following the publication of the above paper, it was drawn to the Editor's attention by concerned readers that β‑actin bands shown in Figs. 1, 2 and 4 were strikingly similar, where the experimental conditions reported in Fig. 4 differed from those in Figs. 1 and 2; moreover, the Slug protein bands featured in Figs. 4a and 5a were remarkably similar in spite of the different experimental conditions that were reported in the respective figure legends, and the shape of the vimentin protein bands in Fig. 5e bore a strong similarity to the Slug protein bands that were featured in Fig. 2c, in spite of the bands being of slightly different sizes and arranged in a different orientation.  Although the possibility of publishing a corrigendum was considered, software analysis of the highlighted bands performed independently by the Editorial Office demonstrated that the bands in question were likely to have been matching bands. Therefore, given the number of potential concerns that were identified with the assembly of various of the figures in this paper, the Editor of International Journal of Oncology has decided not to proceed with a corrigendum, and has determined that the paper should instead be retracted from the Journal on account of an overall lack of confidence in the originally presented data. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a satisfactory reply. The Editor apologizes to the readership for any inconvenience caused. [International Journal of Oncology 46: 1461‑1472, 2015; DOI: 10.3892/ijo.2015.2878].

Hematopoietic stem cells (HSCs) are critical to the development and preservation of the immune system. These scarce cells are distinguished by their remarkable capacity for self-renewal and their potent pluripotency, which allows them to produce all blood cell types, thus maintaining the balance and homeostasis of the hematopoietic system. Their clinical significance extends to regenerative medicine and immunotherapy. Furthermore, HSCs are pivotal in precision medicine, ushering in a new era of monitoring and tracking their reproducibility through methods like liquid biopsy and various immunosurveillance and functional assays. These tests are key for assessing the presence of transplanted cells in the bloodstream or the target organ. Alterations to human cells, whether through the addition, modification, or removal of certain characteristics, can steer them toward becoming fully or partially stem-like cells. This is exemplified by chimeric antigen receptor (CAR) T cells, which are at the forefront of cellular immunotherapy for cancer and are considered a type of engineered stem cells. Nonetheless, disturbances in the development of HSCs caused by genetic or epigenetic mutations, be they inherited or acquired, can compromise immune function, leading to blood-related disorders or cancers. Advances in medical technology, such as improved instruments, reagents, and a deeper understanding of the molecular and biochemical processes of stem cell and HSC development, along with the pathways leading to various immune cell clones, are optimizing the methods for harvesting, culturing, and genetically reprogramming HSCs. These advancements enhance the clinical outcomes for numerous diseases and are transforming the fields of regenerative, precision, and preventive medicine. Within this framework, we are analyzing the connection between normal and dysfunctional HSCs and immunological disorders, focusing on how HSCs are linked with risk factors for genomic instability. These factors include oxidative stress (OS), microsatellite instability (MSI), telomerase activity, the activity of APOBEC enzymes, and epigenetic changes. We are also highlighting the involvement of HSCs in clonal diseases and immune dysfunctions.

Figure S2 A, ERK activation is responsible for FFA promotion of TGF-beta-induced inhibition of SMAD4 monoubiquitination, Related to Figure 2. MDA-MB-231 cells were treated with BSA (vehicle) or PA in the presence or absence of AZD6244 or WP1130 for 4 h followed by treatment with or without 3 ng/ml of TGF-beta for another 4 h. Nuclear extracts were made and SMAD4 monoubiquitination was detected. The blot is a representative of 3 blots from 3 independent experiments. *P<0.01 vs control; #P<0.05, PA vs BSA; $P<0.01, PA+AZD vs PA, PA+WP vs PA). B, ERK activation is responsible for FFA promotion of TGF-beta-induced nuclear SAMD4 retention, Related to Figure 2. MDA-MB-231 cells were treated with 3 ng/ml of TGF-beta in the presence or absence of PA, AZD6244 or WP1130 for the times shown and processed for immunofluorescence with anti-SMAD4 antibody. The same cells were also stained with DAPI to visualize nuclei. Intensity of nuclear SMAD4 among these cells was quantified with Image-Pro Plus 6.0 software. The percentages of nuclear SMAD4 levels illustrated at the lower panel represent the mean of three independent experiments, and error bars indicate the SD.

PDF - 64K, Supplementary Table 4. Evaluation of EMP2 IgG1 toxicity compared to control IgG following 7 weeks of treatment delivered systemically IP. Supplementary Table 5. Evaluation of EMP2 IgG1 toxicity compared to control IgG following systemic IP treatment of up to 40mg/kg.

PDF - 54K, Supplementary Table 2. Clinical Data for Invasive Breast Cancer Patients used in the TMA.

Figure S4 Quantification of Western blot analysis (related to Figure 4) (B-G). Data are presented as mean fold increases ({plus minus}SD) in treated groups over basal values from three independent experiments. B, *p<0.01 vs BSA; #p<0.01 PA+AZD vs PA. E, *p<0.01 vs EV/-PA; #p<0.01 vs EV/+PA. F, *p<0.01 vs EV/-TGF-beta-PA; #p<0.05 vs EV/+TGF-beta-PA; G, *p<0.01 vs WT.

Selected functional pathways enriched with EMP2 mRNA based on TCGA data.

<p>Figure S3 FFA promotes TGF-beta-induced migration by activating ERK and USP9x, Related to Figure 3. A, BT549 cells were plated in Ibidi culture insert dishes (Madison, WI). Cells were cultured in complete medium to 80% confluence, inserts were removed to create a gap and cells were then incubated with BSA, PA, PA plus AZD6244 or WP1130 in the absence or presence of TGF-beta1. Migrating cells were photographed under a phase contrast microscope. B, The percentage of the wound closed was quantified from three independent replicates and is expressed as mean {plus minus} SD. *p<0.05 vs BSA/TGF-beta; #p<0.05 vs PA/TGF-beta.</p>

EMP2 and ALDH1 staining in breast cancer

<p>Figure S5 USP9x is responsible for TGF-beta-induced SMAD4 deubiquitination, Related to Figure 5. MDA-MB-231 cells were transfected with WT-SMAD4, HA-ubiquitin, and the indicated shRNAs and incubated with BSA or PA in the presence or absence of 3 ng/ml TGF-beta1 for 2 h. The effective knockdown of USP9x was confirmed by Western blot analysis before ubiquitination assays. Nuclear SMAD4 monoubiquitination (Mono-Ub) was detected by anti-SMAD4 immunoprecipitation and immunoblot with HA-ubiquitin. The blot is a representative of 3 blots from 3 independent experiments. *P<0.05 vs Control (shControl/-TGF-beta/-PA); #P<0.01 vs shControl/-PA; $P<0.05 shControl/+PA.</p>

Characteristics of 23 mammary and breast cancer cell lines.

<p>Figure S6 The comparison of expression levels of phospho-ERK and phospho-Smad4 between lean and overweight/obesity patients, Related to Figure 7. Staining was analyzed by using a system based on the percentage of positively stained cells and the staining intensity. Integrated optical density of all the positive staining in each image was determined, and its ratio to total area of each photograph was calculated as density.</p>

Supplementary Tables 1-3, Figures 1-2 from Aromatase Expression Predicts Survival in Women with Early-Stage Non–Small Cell Lung Cancer

Selected functional pathways affected by changes in EMP2 expression.

Abstract Little is known about the role of epithelial membrane protein-2 (EMP2) in breast cancer development or progression. In this study, we tested the hypothesis that EMP2 may regulate the formation or self-renewal of breast cancer stem cells (BCSC) in the tumor microenvironment. In silico analysis of gene expression data demonstrated a correlation of EMP2 expression with known metastasis-related genes and markers of cancer stem cells (CSC) including aldehyde dehydrogenase (ALDH). In breast cancer cell lines, EMP2 overexpression increased and EMP2 knockdown decreased the proportion of stem-like cells as assessed by the expression of the CSC markers CD44+/CD24−, ALDH activity, or by tumor sphere formation. In vivo, upregulation of EMP2 promoted tumor growth, whereas knockdown reduced the ALDHhigh CSC population as well as retarded tumor growth. Mechanistically, EMP2 functionally regulated the response to hypoxia through the upregulation of HIF-1α, a transcription factor previously shown to regulate the self-renewal of ALDHhigh CSCs. Furthermore, in syngeneic mouse models and primary human tumor xenografts, mAbs directed against EMP2 effectively targeted CSCs, reducing the ALDH+ population and blocking their tumor-initiating capacity when implanted into secondary untreated mice. Collectively, our results show that EMP2 increases the proportion of tumor-initiating cells, providing a rationale for the continued development of EMP2-targeting agents.

PDF - 51K, Supplementary Table 3. Expression levels of EMP2 in triple negative breast cancer.