Growth Of Breast Carcinoma Cells Research Articles

3,3’4-trimethoxy-4’-rutinosylellagic acid and its acetylated derivative: Antioxidant activity and antiproliferative effects on breast cancer cells and molecular docking study

Cancers account for many deaths worldwide and natural compounds and their derivatives are interesting chemotherapeutic agents for cancer drug development. In this study, a natural compound 3,3’4-trimethoxy-4’-rutinosylellagic acid (TR2) and its acetylated derivative 3,3’4-trimethoxy-4’-hexaacetylrutinosylellagic acid (TR22) were evaluated for their antioxidant and anticancer effects against estrogen sensitive (MCF-7) and estrogen non-sensitive (MDA-MB 231) breast adenocarcinoma. In the β-Carotene-linoleic acid assay, DPPH• radical scavenging and CUPRAC assay, the compound TR2 had better activity than the standard α-Tocopherol, while in the ABTS•+ assay, it was more active than both standards α- α-Tocopherol and BHA. Both compounds had good antioxidant effects with TR2 being more active than TR22. Both compounds inhibited growth of breast carcinoma cells when compared to the untreated controls after 72 h. Compound TR22 significantly (p < 0.001) inhibited proliferation of both MCF-7 and MDA-MB 231 breast carcinoma cell lines suggesting that acetylation reaction improves inhibition of breast cancer cells growth. On the contrary, TR2 exhibited better inhibitory effect of clone formation than TR22 suggesting that acetylation reduces the activity in this assay. Both compounds inhibited migration of the cancer cells when compared to the untreated control cells and compound TR2 exhibited greater cellular anti-migration effect than TR22 at the same concentration and after the same period of incubation. Molecular docking studies supplemented the results and revealed that TR2 and TR22 had appreciable interactions with tyrosine kinase with negative binding energies suggesting that they are potent receptor tyrosine kinase inhibitors which can impede on cancer progression.

In Vitro and In Vivo Antiproliferative Actions of Solanum gilo Raddi (Solanaceae) Fruit Extract on Breast Tissues.

Background Menopause is a normal event characterized by a drop in estrogen's production, leading to numerous symptoms. To face these later, women rely on hormone replacement therapy (HRT), which alleviates numerous menopausal symptoms. Unfortunately, long-term exposure to estrogens is associated with an increase in endometrial and breast cancers. This study dealt with the evaluation of in vitro and in vivo antiproliferative effects of Solanum gilo Raddi, a plant used in folk medicine to treat tumors in Cameroon. Materials and Methods The in vitro antiproliferative effect of S. gilo fruit extract was investigated through the well-characterized MTT assay in one normal and three cancerous breast cells. For the in vivo study, one normal group (NOR) of rats received distilled water (vehicle), and five other groups (n = 6) were treated either with tamoxifen (3.3 mg/kg BW) as standard or with the vehicle (negative control) or S. gilo fruit hydroethanolic extract (125, 250, and 500 mg/kg BW). The treatments were administered concomitantly with the E2V to induce breast hyperplasia for 16 weeks, and the endpoints were the histopathology of the mammary glands and some biochemical parameters. Results The S. gilo extract significantly inhibited human (MCF-7 and MDA-MB-231) and rodent (4T1) breast carcinoma cell growth. Rats exposed only to E2V presented atypical mammary hyperplasia compared to the normal parenchyma observed in normal rats. While rats treated with S. gilo extract at the dose of 125 mg/kg BW showed a microarchitecture of mammary glands with moderate hyperplasia, the higher doses (250 and 500 mg/kg) inhibited mammary gland hyperplasia compared to the E2V group. Conclusion S. gilo fruit extract has antiproliferative constituents that could help to fight against estrogen-dependent breast cancer, thanks to their ability to scavenge free radicals, as exhibited in this study.

MiR-450a exerts oncosuppressive effects in breast carcinoma by targeting CREB1.

Emerging evidence greatly implicates that microRNA-450a (miR-450a) plays an essential role in cancer pathobiology. While the pathological role of miR-450a in breast carcinogenesis remains enigmatic. Herein, we showed that miR-450a was lowly expressed in breast cancer cell lines compared with normal, and low miR-450a expression was associated with poor survival in patients with breast cancer. We revealed that miR-450a mimic transfected breast cancer cells (T47D and BT474) exhibited attenuated capacities of proliferation, migration, and invasion in vitro, and miR-450a suppressed T47D cell growth in a xenograft tumor model. Mechanistically, cAMP response element-binding protein 1 (CREB1) was negatively targeted by miR-450a, and CREB1 deletion mimicked the effects of miR-450a mimic treatment. Bioinformatics analysis further revealed that elevated expression of CREB1 correlated with poor prognosis in patients with breast cancer and miR-450a level was negatively correlated with CREB1 level in breast cancer. Additionally, miR-450a inhibited the phosphorylation of phosphatidylinositol 3-kinase/V-akt murine thymoma viral oncogene homolog (PI3K/AKT) and the activities of matrix metalloproteinase-2/9 (MMP-2/9). The following rescue assay indicated that CREB1 was implicated in the anti-tumoral effect of mR-450a in breast carcinoma. All these observations disclosed that miR-450a negatively regulates the growth and metastatic property of breast carcinoma cells.

Synthesis, characterization, and biological activity of Co(II) and Zn(II) complexes of imidazoles-based azo-functionalized Schiff bases

A new series of azo-functionalized Schiff base ligands containing different imidazole derivatives (HL1‒HL3) and their Co(II)/Zn(II) complexes were successfully prepared. The molecular structures of the new ligands and their metal complexes have been deduced using microanalytical and spectral techniques. The thermal stability study revealed that the Co(II)/Zn(II) complexes are more stable than their corresponding ligands. The results of in vitro cytotoxicity test revealed that all compounds have the capacity to inhibit the growth of breast carcinoma (MCF-7) cells with a structure-dependent performance. Among all, complex CoL3, (IC50 = 0.90±0.05 µg/mL) was the most potent and selective (SI = 90.56) anti-breast cancer agent. Flow-cytometric study revealed that CoL3 dramatically enhanced the proportion of MCF-7 cells in the sub G1 phase while decreasing the proportion of cells in the S phase. In addition, CoL3 showed better inhibitory activity on DNA topoisomerase-II.

Exhausting hsa_circ_0072088 restrains proliferation, motility and angiogenesis of breast carcinoma cells through regulating miR-1236-3p and RRM2 in a ceRNA pathway

BackgroundHsa_circ_0072088 (circ_0072088) is a novel and informative circular RNA in several human cancers, including breast carcinoma (BRCA). Herewith, we aimed at investigating the role and mechanism of circ_0072088 in BRCA cells. MethodsExpression of circ_0072088, microRNA (miR)-1236-3p, ribonucleotide reductase subunit M2 (RRM2), and epithelial-to-mesenchymal transition (EMT)-related markers E-cadherin, N-cadherin and Vimentin was detected by quantitative PCR and/or western blotting. Cell proliferation was measured by assays using MTS, EdU, colony formation, immunohistochemistry and nude mice tumorigenicity. Migration and/or invasion and angiogenesis were determined by scratch wound assay, transwell assay, and Capillary-like tube formation assay. Intermolecular interdependence was determined by bioinformatics algorithms, dual-luciferase reporter assay, RNA pull-down assay and Pearson's correlation coefficient analysis. ResultsCirc_0072088 was upregulated in BRCA patients’ tumors, and its high expression was associated with clinical metastasis and unfavorable disease-free survival. Interfering circ_0072088 could weaken cell viability, colony formation, EdU incorporation, and tumor growth of BRCA cells; moreover, circ_0072088 interference suppressed wound healing, migration/invasion and EMT process of BRCA cells, and tube formation of endothelial cells. Strikingly, either miR-1236-3p inhibitor or RRM2 overexpression could significantly abate in vitro roles of circ_0072088 blockage. Allied with that, circ_0072088 was a sponge of miR-1236-3p in regulating expression of miR-1236-3p-specific RRM2. ConclusionInhibiting circ_0072088 might treat BRCA via miR-1236-3p/RRM2 axis, suggesting circPHKA2 as a novel molecular inhibitor of RRM2, a Food and Drug Administration (FDA)-approved drug target.

Role of miR-100-5p and CDC25A in breast carcinoma cells.

ObjectiveTo inquiry about mechanism of miR-100-5p/CDC25A axis in breast carcinoma (BC), thus offering a new direction for BC targeted treatment.MethodsqRT-PCR was employed to explore miR-100-5p and CDC25A mRNA levels. Western blot was employed for detecting protein expression of CDC25A. Targeting relationship of miR-100-5p and CDC25A was verified by dual-luciferase assay. In vitro experiments were used for assessment of cell functions.ResultsIn BC tissue and cells, miR-100-5p was significantly lowly expressed (P < 0.05) while CDC25A was highly expressed. Besides, miR-100-5p downregulated CDC25A level. miR-100-5p had a marked influence on the prognosis of patients. The forced miR-100-5p expression hindered BC cell proliferation, migration and invasion, and facilitated cell apoptosis. Upregulated miR-100-5p weakened promotion of CDC25A on BC cell growth.ConclusionTogether, these findings unveiled that CDC25A may be a key target of miR-100-5p that mediated progression of BC cells. Hence, miR-100-5p overexpression or CDC25A suppression may contribute to BC diagnosis.

Mediation of circ_RPPH1 on miR-146b-3p/E2F2 pathway to hinder the growth and metastasis of breast carcinoma cells.

Background: Nova Circular RNA (circRNA) of non-coding RNA has gradually become an important regulatory factor, and it has made people attach great concern over the occurrence and development of many diseases, particularly carcinomas. circ_RPPH1 is a newly discovered circRNA. Gene Expression Omnibus (GEO) analysis showed that there are high contents of circ_RPPH1 in breast cancer (BC), but the mechanism of circRNA in BC remains unclear.Methods: Real-time quantitative PCR (qRT-PCR) was applied to test the role of circ_RPPH1 in BC patients, and functional experiments were applied to test the role of circ_RPPH1 on BC tumor. Fluorescence in situ hybridization, double luciferase reporter gene analysis, RNA pull-down and RNA immunoprecipitation experiments were performed to explore the correlation of circ_RPPH1 with miR-146b-3p/E2F2 in BC.Results: circ_RPPH1 was evidently enhanced in BC, and its content was related to the clinical stage and pathological grade. circ_RPPH1 can accelerate the proliferation, migration and invasion, and promote tumorigenesis and metastasis. Mechanism exploration indicated that circ_RPPH1 acted as ceRNA (competing endogenous RNA) of miR-146b-3p, so as to reduce the inhibitory role of miR-146b-3p on its target E2F2.Conclusion: Circ_RPPH1/miR-146b-3p/E2F2 axis can promote the progression of BC, and it might be a latent therapeutic target for clinical BC.

The Extracts of Epilobium Parviflorum Inhibit MCF-7 Breast Cancer Cells

Background: Traditional medicine is inspiring in drug development research. Epilobium parviflorum extracts have shown promising therapeutic effects on prostate cancer cells. The similarities between breast and prostate cancers at molecular and metabolic levels prompted us to explore its effects on human breast cancer. Methods: The root, aerial parts and flowers of the plant were, collected and dried separately at ambient temperature and away from direct sunlight. The aquatic and methanolic extracts of each part was prepared. The effect of each extract on the growth of MCF-7 breast carcinoma cells and HEK293 normal cell line was evaluated, using MTT assay. Each experiment was repeated at least three times independently. The IC50 values for each treatment time point were analyzed, using ANOVA at P&lt;0.05. Results: While none of the extracts had considerable toxicity on normal HEK293 cells, some of them showed varying levels of toxicity on the MCF-7 cells. The methanolic extracts were more cytotoxic than the aqueous counterpart. The roots’ methanolic extract showed the strongest cytotoxicity on the MCF-7 cells in a dose and exposure time dependent manner. The IC50 after 48 hours of treatment was determined at 73µg/ml. Conclusion: This study is the first to demonstrate that Epilobium parviflorum had a strong growth inhibiting property on MCF-7 cell line, as a potential model to treat human breast cancer cells. The most cytotoxic effect was noted for the methanolic root extract. Determination of the effective biochemical constituents of the extract against cancer cells is the focus of our future research.

LncRNA ADAMTS9-AS1 Restrains the Aggressive Traits of Breast Carcinoma Cells via Sponging miR-513a-5p.

PurposeLong noncoding RNAs (lncRNAs) exert important functions in the progression of cancers. Currently, we aim to investigate the potential roles of lncRNA ADAM Metallopeptidase with Thrombospondin Type 1 Motif 9 Antisense RNA 1 (ADAMTS9-AS1) in breast carcinoma.Materials and MethodsThe expressions of ADAMTS9-AS1 and miR-513a-5p in breast carcinoma tissues and cell lines were detected using qRT-PCR. Cell Counting Kit-8 (CCK-8) and transwell assays were used to assess the viability and invasive ability of breast cancer cells. The direct interaction between ADAMTS9-AS1 and miR-513a-5p was predicted using bioinformatics tools. The target of miR-513a-5p, ZFP36 Ring Finger Protein (ZFP36) was validated by luciferase assay. The expression of ZFP36 was measured using Western blot assay. Breast cancer MDA-MB-231 cells growth in vivo was evaluated using xenograft tumor assay.ResultsADAMTS9-AS1 was downregulated in breast cancer tissues as well as cell lines. Upregulation of ADAMTS9-AS1 suppressed the growth and invasiveness of breast carcinoma cells in vitro as well as inhibiting cellgrowth in vivo. Furthermore, ZFP36 was manifested as the target gene of miR-513a-5p and negatively modulated by ADAMTS9-AS1. In addition, overexpression of ADAMTS9-AS1 neutralized the promoting impact of miR-513a-5p on the aggressiveness of breast cancer cells.ConclusionIn conclusion, lncRNA ADAMTS9-AS1 inhibited the aggressive phenotypes of breast carcinoma cells via sponging miR-513a-5p and regulating ZFP36.

CYP24A1‐induced vitamin D insufficiency promotes breast cancer growth

Vitamin D plays a critical role in the maintenance of tissue homeostasis by regulating the expression of genes affecting cell proliferation, differentiation, and apoptosis. The vitamin D 24‐hydroxylase CYP24A1 functions in vitamin D target tissues to degrade the hormonal form of vitamin D. Existing knowledge regarding dysregulated CYP24A1 expression supports its candidacy as a putative oncogene. In our presentation, we demonstrate that the suppression of constitutive CYP24A1 expression by CYP24A1‐specific shRNA conferred target cells with increased susceptibility to apoptosis and consequently inhibited anchorage‐independent growth in breast carcinoma cells. In addition, suppression of vitamin D metabolism following knockdown of CYP24A1 significantly reduced tumor growth in vivo. Since these data provide substantial evidence for a pro‐survival and stimulatory oncogenic effect of CYP24A1 in breast carcinoma cells, we will discuss the potential feasibility of CYP24A1‐inhibitory cancer therapies.

Oncogenic microRNA-765 promotes the growth and metastasis of breast carcinoma by directly targeting ING4.

Previous investigations have proved that microRNA (miR)-765 is significantly overexpressed in multiple tumor types. Nevertheless, the underlying molecular mechanism of miR-765 in mediating breast carcinoma cell growth and metastasis remains unclear. Quantitative real-time polymerase chain reaction was used to determine the levels of miR-765 and inhibitor of growth 4 (ING4) in breast carcinoma tissues and breast carcinoma cells. Cell proliferation, colony formation, wound healing, and Transwell invasion assays were used to analysis the role of miR-765 on breast carcinoma cell growth and aggressiveness. The expressions of ING4 were determined using Western blot analysis and immunohistochemical staining. The direct target of ING4 and miR-765 was confirmed using the luciferase reporter assay. Nude mice were subcutaneously implanted with miR-765 inhibitor transfected MDA-MB-231 cells to determine the potential role of miR-765 in tumor growth in vivo. We observed that miR-765 is overexpressed in breast carcinoma tissue and breast cancer cells. By using luciferase reporter gene bioassay, we find that ING4 is the direct target of miR-765 in breast carcinoma. The level of ING4 is inversely associated with the level of miR-765. The gain-of-function and loss-of-function experiments in vitro indicate that thedownregulation of miR-765 suppresses the growth, mobility, and invasion abilities of breast cancer cells by inhibiting ING4. In addition, overexpression of ING4 suppresses the aggressiveness of the MDA-BA-231 cell that is induced by miR-761 in vitro. In this study, we prove that miR-765 regulates the growth and metastasis of breast cancer via modulating miR-765-ING4-negative feedback loop.

Danshen Improves Survival of Patients With Breast Cancer and Dihydroisotanshinone I Induces Ferroptosis and Apoptosis of Breast Cancer Cells.

Danshen (salvia miltiorrhiza Bunge) is widely used in traditional Chinese medicine. However, it is definite clinical effort and mechanism on breast cancer is unclear. In our study, we used the real-world database to investigate in vivo protective effort of danshen in the breast cancer patients through using population-based data from the Taiwan National Health Insurance Research Database (NHIRD). In vitro, human breast cancer cells (MCF-7 cells and MDA-MB-231 cells) were used to investigate the effect and the underlying mechanism through XTT assay, flow cytometry, glutathione peroxidase (GPX) activity assay, GSH (reduced glutathione)/GSSG (oxidized glutathione), malondialdehyde (MDA), and western blot analysis. The in vivo effect was investigated through a xenograft nude mouse model. We found that dihydroisotanshinone I (DT), a pure compound present in danshen, can inhibit the growth of breast carcinoma cells, including MCF-7 cells and MDA-MB-231 cells. Moreover, DT induced apoptosis and ferroptosis in these breast cancer cells. DT also repressed the protein expression of GPX4 (Glutathione peroxidase 4). For in vivo study, DT treatment also significantly inhibited the final tumor volume without adverse effects in a xenograft nude mouse model. In conclusion, danshen has protective efforts in breast cancer patients, which could be attributed to DT through inducing apoptosis and ferroptosis of breast cancer cells.

Cyanidin-3-O-glucoside chloride acts synergistically with luteolin to inhibit the growth of colon and breast carcinoma cells.

In this study, we investigated whether the combination of cyanidin-3-O-glucoside chloride and luteolin could inhibit the growth of HCT-8 colon and MCF-7 breast carcinoma cells, and whether the effect of the combination was greater than the effect of either drug on its own. Growth inhibition was assessed using the CCK-8 assay, level of apoptosis and cell cycle distribution were determined using flow cytometry, and the mechanism of apoptosis induction was explored using a colorimetric assay of caspases-3 and -9. Experiments indicated that combination treatment inhibited proliferation and increased apoptosis in both cell lines to a greater extent than either drug on its own. These results suggest that luteolin and cyanidin-3-O-glucoside chloride synergistically inhibitthe growth of colon cancer and breast cancer cells. This work justifies further effort to develop this potential combination therapy.

Abstract 2485: CYP24A1-induced vitamin D insufficiency promotes breast cancer growth

Abstract Vitamin D plays a critical role in the maintenance of tissue homeostasis by regulating the expression of genes affecting cell proliferation, differentiation, and apoptosis. The vitamin D 24-hydroxylase CYP24A1 functions in vitamin D target tissues to degrade the hormonal form of vitamin D. Existing knowledge regarding dysregulated CYP24A1 expression supports its candidacy as a putative oncogene. In our presentation, we demonstrate that the suppression of constitutive CYP24A1 expression by CYP24A1-specidic shRNA conferred target cells with increased susceptibility to apoptosis and consequently inhibited anchorage-independent growth in breast carcinoma cells. In addition, suppression of vitamin D metabolism following knockdown of CYP24A1 significantly reduced tumor growth in vivo. Since these data provide substantial evidence for a pro-survival and stimulatory oncogenic effect of CYP24A1 in breast carcinoma cells, we will discuss the potential feasibility of CYP24A1-inhibitory cancer therapies. Citation Format: Makoto Osanai, Yusuke Ono, Akira Takasawa, Kumi Takasawa, Masaki Murata, Norimasa Sawada. CYP24A1-induced vitamin D insufficiency promotes breast cancer growth [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2485.

Sema4C/PlexinB2 signaling controls breast cancer cell growth, hormonal dependence and tumorigenic potential.

Semaphorin 4C (Sema4C) expression in human breast cancers correlates with poor disease outcome. Surprisingly, upon knock-down of Sema4C or its receptor PlexinB2 in diverse mammary carcinoma cells (but not their normal counterparts), we observed dramatic growth inhibition associated with impairment of G2/M phase transition, cytokinesis defects and the onset of cell senescence. Mechanistically, we demonstrated a Sema4C/PlexinB2/LARG-dependent signaling cascade that is required to maintain critical RhoA-GTP levels in cancer cells. Interestingly, we also found that Sema4C upregulation in luminal-type breast cancer cells drives a dramatic phenotypic change, with disassembly of polarity complexes, mitotic spindle misorientation, cell-cell dissociation and increased migration and invasiveness. We found that this signaling cascade is dependent on the PlexinB2 effectors ErbB2 and RhoA-dependent kinases. Moreover, Sema4C-overexpressing luminal breast cancer cells upregulated the transcription factors Snail, Slug and SOX-2, and formed estrogen-independent metastatic tumors in mice. In sum, our data indicate that Sema4C/PlexinB2 signaling is essential for the growth of breast carcinoma cells, featuring a novel potential therapeutic target. In addition, elevated Sema4C expression enables indolent luminal-type tumors to become resistant to estrogen deprivation, invasive and metastatic in vivo, which could account for its association with a subset of human breast cancers with poor prognosis.

β-inducible gene-h3 promotes human breast carcinoma cell metastasis by activating the phosphatidylinositol 3-kinase/protein kinase B signaling pathway.

Metastatic breast cancer is one of the most common metastatic tumors. Although studies have validated the role of β-inducible gene-h3 (βig-h3) in human biology and disease, the detailed mechanisms mediated by βig-h3 in breast carcinoma metastasis remain unclear. Thus, the present study investigated the role and potential mechanism of βig-h3 during breast carcinoma cell metastasis. The results indicated that the upregulation of βig-h3 significantly promotes the growth and inhibits the cisplatin-induced apoptosis of breast carcinoma cells. It was also demonstrated that βig-h3 promoted the migration and invasion of human breast carcinoma cells in vitro and in vivo. Furthermore, the results demonstrated that βig-h3 upregulated the overall expression and phosphorylation of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt) in human breast carcinoma cells. By contrast, βig-h3 knockdown reversed the βig-h3-mediated characteristics of breast carcinoma cells. Thus, the current study demonstrated that the PI3K/Akt signaling pathway serves a role in βig-h3-induced human breast cancer cell metastasis and that βig-h3 transfection enhances the metastatic potential of human breast carcinoma cells via the PI3K/Akt signaling pathway. These observations contribute to the understanding of the potential mechanism of human breast carcinoma cell growth and metastasis and suggest that βig-h3 may be a promising therapeutic target for the treatment of human breast carcinoma.

Abstract: Analysis of Proinflammatory Markers in Lipofilled Breast Microenvironment in a Murine Model

INTRODUCTION: Fat grafting to the breast has a questionable oncologic risk according to laboratory reports on adipose tissue. One possible reason for this is the theoretical chronic inflammation due to adipokynes released by the grafted white adipose tissue (WAT).1–4 Thus, a translational study was designed to analyze inflammatory activity in lipofilled breast through proinflammatory markers. METHODS: Fifty-four paired-breasts of female rats were divided into three groups (control, grafted with autologous WAT of subcutaneous, and of omentum). The procedures of WAT preparation and grafting were performed as described previously.1–5 Briefly, a piece of WAT was harvested from omentum or subcutaneous and manually chopped with a scalpel into very small pieces and then placed into a 10-ml syringe. The WAT was then transferred into another 10-ml syringe through an emulsifier; in this way, the samples were homogeneously fragmented. Next, the samples were centrifuged at 3,000 rpm for 3 minutes. The lower and uppermost levels were discharged, such that only the middle layer (containing viable cells) was transferred into a 1-ml syringe. Finally, 0.2 ml of prepared autologous WAT (from omentum or subcutaneous tissue) was grafted through a 0.9-mm diameter cannula with a blunt tip into each of the 6 thoracic breasts of each rat. The rats were euthanized at 8 weeks post-operatively, and their breasts were harvested for analysis of gene quantification (via real-time PCR) for Mcp-1, F4/80, Cox-2 and IL-6. Statistical analyses were performed using paired t-tests, analysis of variance (ANOVA, one way) and Tukey’s test (post hoc). RESULTS: The markers Mcp-1, F4/80 and Cox-2 were similarly expressed among the groups (respectively p=0.422, p=0.143 and p=0.209). The expression of IL-6 marker was different between samples of breast grafted with WAT of omentum and subcutaneous (p=0.015), but not between samples of control and omentum (p=0.752), and control and subcutaneous (p=0.056). CONCLUSION: The difference in fold-change for IL-6 only between groups of lipofilled breasts, but not with control, certainly resulted from the different concentration of WAT of omentum and subcutaneous (they are from different embryological origin) in the breast composition in each group. Our results showed that no inflammation activity was identified in the lipofilled breast microenvironment, this illustrates that the tumorigenic potential of WAT reported for the mammary gland (regarding chronic inflammation) was not identified in our study. Reference Citations: 1. Claro, F., Jr., Moreira, L. R., Morari, J., et al. Assessment of the Cancer Risk of the Fat-Grafted Breast in a Murine Model. Aesthet Surg J 2016. 2. Gutowski, K. A., Force, A. F. G. T. Current applications and safety of autologous fat grafts: a report of the ASPS fat graft task force. Plast Reconstr Surg 2009;124:272–280. 3. Claro Jr, F., Figueiredo, J. C. A., Zampar, A. G., Pinto-Neto, A. M. Applicability and safety of autologous fat for reconstruction of the breast. British Journal of Surgery 2012;99:768–780. 4. Manabe, Y., Toda, S., Miyazaki, K., Sugihara, H. Mature adipocytes, but not preadipocytes, promote the growth of breast carcinoma cells in collagen gel matrix culture through cancer–stromal cell interactions. Journal of Pathology 2003;201:221–228. 5. Coleman, S. R. Structural fat grafting. St. Louis: Quality Medical Publishing; 2004.

Abstract 2567: CYP24A1-induced vitamin D insufficiency promotes breast cancer growth

Abstract Vitamin D plays a critical role in the maintenance of tissue homeostasis by regulating the expression of genes affecting cell proliferation, differentiation, and apoptosis. The vitamin D 24-hydroxylase CYP24A1 functions in vitamin D target tissues to degrade the hormonal form of vitamin D. Existing knowledge regarding dysregulated CYP24A1 expression supports its candidacy as a putative oncogene. In our presentation, we demonstrate that the suppression of constitutive CYP24A1 expression by CYP24A1-specidic shRNA conferred target cells with increased susceptibility to apoptosis and consequently inhibited anchorage-independent growth in breast carcinoma cells. In addition, suppression of vitamin D metabolism following knockdown of CYP24A1 significantly reduced tumor growth in vivo. Since these data provide substantial evidence for a pro-survival and stimulatory oncogenic effect of CYP24A1 in breast carcinoma cells, we will discuss the potential feasibility of CYP24A1-inhibitory cancer therapies. Citation Format: Makoto Osanai, Akira Takasawa, Kumi Takasawa, Masaki Murata, Norimasa Sawada. CYP24A1-induced vitamin D insufficiency promotes breast cancer growth [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 2567. doi:10.1158/1538-7445.AM2017-2567

Interaction with adipocyte stromal cells induces breast cancer malignancy via S100A7 upregulation in breast cancer microenvironment

BackgroundBreast adipocytes play important roles in both the development and function of mammary epithelial cells. Therefore, carcinoma–adipose stromal cell (ASC) interactions have been considered pivotal in supporting tumor growth in breast cancer. In addition, it has been demonstrated that the biological features of cancer-associated adipocytes differ from those of normal ASCs. Therefore, we investigated an interaction between ASCs and carcinoma cell lines to identify genes associated with ASC invasion of carcinoma cells.Methods3T3-L1 ASC-derived conditioned medium (CM) was treated to measure the proliferation rate of breast cancer cells. To further examine the effect of ASCs, breast cancer cells were cocultivated with either primary human or 3T3-L1 ASCs for migration assays, DNA microarrays, quantitative real-time polymerase chain reactions, and Western blotting experiments. Furthermore, immunoreactivity of S100A7, the most upregulated gene in MCF7, after coculture with ASCs was evaluated for 150 breast cancer tissues to statistically analyze its association with clinicopathological parameters.ResultsWe first confirmed that ASC-derived CM treatment enhanced the cell proliferation rate of MCF7, T47D, SK-BR-3, and ZR-75-1 cell lines, whereas the migration rate of breast cancer cells was promoted by coculture with ASCs. We identified that a small calcium-binding protein, S100A7, was markedly upregulated (by 5.8-fold) in MCF7 cells after coculture with primary human ASCs. Knockdown of S100A7 significantly suppressed ASC-stimulated cell proliferation and migration rate, indicating a possible involvement of S100A7 in the carcinoma–ASC interaction in breast tumors. Furthermore, strong S100A7 immunoreactivity was detected at the invasive front of adipose stromal tissues compared with that at the intratumoral area. The status of S100A7 was also significantly correlated with adverse pathological parameters, and multivariate analysis revealed that S100A7 could be an independent prognostic marker for a poor relapse-free survival rate. Moreover, induction of oncostatin M was detected in cancer-stimulated ASCs, whereas the downstream S100A7 binding proteins/receptor for advanced glycation endproducts were significantly upregulated in correspondence with S100A7 expression in breast cancer cells after coculture with ASCs.ConclusionsThe results of our study suggest that paracrine production of cytokines from ASCs stimulates breast carcinoma cell growth via upregulation of S100A7 expression in breast cancer cell lines.

Identification of Novel Human Breast Carcinoma (MDA-MB-231) Cell Growth Modulators from a Carbohydrate-Based Diversity Oriented Synthesis Library.

The application of a cell-based growth inhibition on a library of skeletally different glycomimetics allowed for the selection of a hexahydro-2H-furo[3,2-b][1,4]oxazine compound as candidate inhibitors of MDA-MB-231 cell growth. Subsequent synthesis of analogue compounds and preliminary biological studies validated the selection of a valuable hit compound with a novel polyhydroxylated structure for the modulation of the breast carcinoma cell cycle mechanism.