This investigation identifies several potential molecular targets of crucifers that may aid in studying established and emerging health benefits of consuming cruciferous vegetables and related bioactive compounds.
Objectives Triple negative breast cancer (TNBC) accounts for 12% to 24% of all breast cancer cases and is characterized by higher proliferation rates and an increased likelihood of tissue invasion. Tumor cells and cells in the tumor microenvironment (TME), specifically tumor associated macrophages (TAMs), interact through signaling (e.g., cytokines) to promote cancer progression by increasing proliferation and invasion capacity. Sulforaphane (SFN) is an isothiocyanate derived from cruciferous vegetables (i.e., broccoli) that has shown promising results in hindering TNBC progression. SFN reduced proliferation in breast cancer cells and altered cytokine signaling between breast cancer cells and cells in the TME (adipocytes). SFN also decreased invasion markers in TNBC cells grown in isolation. However, TAMs promote tumor cell aggression, and SFN's effect in multicellular environments is unclear. To determine SFN's potential in cancer treatment, it is critical to investigate SFN's effect on proliferation and invasion capacity of TNBC cells grown under TAMs influence, not just grown in isolation. The objective of this study was to determine if SFN can reduce proliferation and invasion capacity of TNBC cells grown in TAM secretions. Methods For cell proliferation, TNBC cells (MDA-MB-231) were exposed to TAM secretions using conditioned media and were then treated with SFN (10 μM) or DMSO vehicle control for 24 and 48 hours. Proliferation was measured using a MTT-based assay and cell counts. For invasion, TAMS and TNBC cells were co-cultured for 48 hours in transwell plates. Prior to co-culture period, cells were treated with SFN (15 μM) or vehicle control. Invasion capacity was measured through a transwell invasion assay with collagen representing the tumor basement membrane. ANOVA and t-tests were used to determine statistical differences with significance at P = 0.05. Results Preliminary analysis revealed significant reductions in proliferation for TAM exposed TNBC cells after 24 hours (P = 0. 0132) and 48 hours (P = 0. 0190) of SFN treatment. Conclusions SFN reduced proliferation and can potentially reduce invasion capacity of TNBC cells influenced by TAM secretions thus enhancing future utilization of SFN in TNBC treatment regimens. Funding Sources California State University, Chico.
Objectives Triple-negative breast cancer (TNBC) comprises 10–20% of breast cancer cases. It is particularly aggressive with limited and deleterious treatment options. Increasingly, research confirms that communication between cancer cells and neighboring macrophages promotes disease progression in part by secretion of cytokines that increase tumor cell proliferation, invasion, and metastasis. Sulforaphane (SFN) is a chemopreventive phytochemical found in cruciferous vegetables (broccoli) shown to alter cytokine secretion in macrophages and breast cancer cells grown in single culture. However, its effect in the tumor microenvironment remains unclear. This study aims to characterize cytokine profiles in media where TNBC cells and macrophages are grown in coculture with and without SFN treatment. We expect SFN to modify cytokine secretions in coculture media, suggesting SFN may disrupt vital cell-cell signaling needed for cancer progression. Methods TNBC cells (MDA-MB-231) were grown in Transwell plates with and without macrophages (THP-1 cells differentiated with PMA). Cell cultures (n = 3) were treated with either 15 μM SFN, DMSO (vehicle-control), or a non-treatment control. Cytokine levels were evaluated in media at 24 and 48 hours after treatment using BioPlex 2000 assay. Results Treatment with sulforaphane significantly reduced the levels of several targets in coculture including IL-1ra, IL-4, IL-5, IL-10, IL-12, IL-13, IL-15, IL-17, CCL2 (MCP-1), CCL11, CCL22, CCL26, CXCL12, IFN-y, G-CSF, GM-CSF, Eotaxin, and VEGF. Conversely, MIF was elevated following treatment. Effects were discovered at 24-hour and 48-hour time points. Conclusions We demonstrated that SFN altered the levels of numerous cellular signaling proteins in cancer cell-macrophage coculture, many of which are known to be involved with breast cancer progression. These results reveal mechanistic links underlying SFNs chemopreventive function and bolster SFNs potential as a treatment strategy for TNBC. Funding Sources Department of Nutrition and Food Science, CSU Chico; Graduate Studies, CSU Chico; CSUPERB: CSU Program for Education and Research in Biotechnology.
Objectives Triple negative breast cancer (TNBC) makes up approximately 10–20% of all breast cancer cases and is more common in younger women and in Hispanic and African American populations. It is particularly difficult to treat, exhibiting high-metastasis rates, poor prognosis, and limited treatment options. Mortality from TNBC is largely due to the tumor cells high invasive capacity and rapid progression to metastasis. Evidence suggests that macrophages in the breast tumor microenvironment release cytokines that increase tumor cell proliferation, invasion and metastasis. Sulforaphane (SFN) is a broccoli phytochemical that has been identified to slow the progression of breast cancer as well as alter cytokine secretion from macrophages and breast cancer cells grown in single culture. SFN effects on cytokine secretion in the breast tumor microenvironment remain unclear. This study is investigating the effect of SFN on cytokine levels in cell culture media of TNBC cells grown with and without macrophages. Our hypothesis is that cytokine levels differ in media from cocultured cells versus singly cultured cells, and SFN treatment further alters cytokine levels in media. Methods In this study, TNBC cells (MDA-MB-231) were grown in transwell plates with and without macrophages (THP-1 cells differentiated with phorbol-myristate acetate). Cell cultures (n = 3) were treated with either 15 µM SFN, DMSO (vehicle-control), or a non-treatment control. We evaluated the levels of 44 individual cytokines in cell culture media at 24 and 48 hours after treatment using a multi-plex (BioPlex) assay. Control groups included single-cultured MDA-MB-231 and differentiated THP-1 cells. Results Preliminary analyses revealed that cytokine levels differed in the media of single versus cocultured cells and among treatment groups after 24 and 48 hours of treatment. Conclusions The profile of cytokines in the media of TNBC cells grown with macrophages was influenced by SFN treatment. This information may help establish mechanisms underlying SFN effects on TNBC behavior and identify new treatment strategies. Funding Sources California State University Program for Education and Research in Biotechnology, California State University-Chico.
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