Evaluating the Effects of an Organic Extract from the Mediterranean Sponge Geodia cydonium on Human Breast Cancer Cell Lines

Abstract

Marine sponges are an excellent source of bioactive secondary metabolites for pharmacological applications. In the present study, we evaluated the chemistry, cytotoxicity and metabolomics of an organic extract from the Mediterranean marine sponge Geodia cydonium, collected in coastal waters of the Gulf of Naples. We identified an active fraction able to block proliferation of breast cancer cell lines MCF-7, MDA-MB231, and MDA-MB468 and to induce cellular apoptosis, whereas it was inactive on normal breast cells (MCF-10A). Metabolomic studies showed that this active fraction was able to interfere with amino acid metabolism, as well as to modulate glycolysis and glycosphingolipid metabolic pathways. In addition, the evaluation of the cytokinome profile on the polar fractions of three treated breast cancer cell lines (compared to untreated cells) demonstrated that this fraction induced a slight anti-inflammatory effect. Finally, the chemical entities present in this fraction were analyzed by liquid chromatography high resolution mass spectrometry combined with molecular networking.

Keywords: breast cancer; cytokines; cytotoxicity; metabolomics; sponges.

Figure 1

Cell proliferation. Cell viability rate (CR) related to breast cancer cells: (a) MCF-7; (b) MDA-MB231; and (c) MDA-MB468, after treatment with the active sponge Geodia cydonium sub-fraction for 24 (blue line) and 48 (red line) h.

Figure 2

(Left) Two-dimensional molecular network of the active fraction from the sponge Geodia cydonium. In the clusters indicated with (a–f), nodes are labeled with parent m/z ratio (M+H)⁺ ions; edge thickness is related to cosine similarity score; (Right) Liquid chromatography-high resolution mass spectrometry (LC-HRMS) profile of the active fraction; the most abundant ions are represented by colored bold contour squares.

Figure 3

RT-qPCR analysis: mRNA fold changes were evaluated as ratios between the expression levels of five genes in three breast cancer cell lines, MCF-7, MDA-MB231 and MDA-MB468, after treatment with the active fraction compared to untreated cells.

Figure 4

Orthogonal Projections to Latent Structures discriminant analysis (OPLS-DA) plots: (A) OPLS-DA and variable importance in projection (VIP) analysis where the metabolites increased or decreased in the endo-metabolome of: (B) MCF-7; (C) MDA-MB231; and (D) MDA-MB468 cells after treatment with the active fraction from the sponge Geodia cydonium, compared to untreated cells.

Figure 5

Cytokine levels in breast cancer cells. Scheme reporting the cytokines modulated by the active fraction obtained from the sponge Geodia cydonium. In particular, anti-inflammatory cytokines that are increased after treatment are reported in red and pro-inflammatory and pro-angiogenic cytokines that are decreased after treatment are reported in green. Blank cells indicate that the cytokines are not modulated by active fraction.