Anti-metastatic potential of anthocyanins and their metabolites (metabotypes) on cancer cell lines with different phenotypes - role of intestinal fermentation and metabolization.

在德国，癌症约占20 - 30%，是第二大死亡原因。肿瘤的侵袭性取决于其转移潜力，其特征在于侵袭、内渗和外渗的步骤。新的药理学治疗剂特别旨在抑制肿瘤细胞的细胞粘附分子，例如整合素、选择素和免疫球蛋白超家族的成员，其表达和模式与转移潜能强烈相关。它们旨在抑制肿瘤细胞外渗到组织中和转移的形成。目前正在深入研究能够影响这些过程的营养因素。首先，花青素（ACN）属于植物次生成分，具有特别重要的意义。它们的生物利用度很低，并且它们到达下胃肠道时基本上没有变化。在这里，可能会产生微生物发酵产物，这些产物在被吸收到体循环中后可以诱导抗癌作用。初步实验表明，由典型存在于人类肠道微生物群中的细菌如E。coli和Haf. alvei显著降低结肠直肠癌和胰腺癌细胞的体外迁移。同样，摄入富含ACN的果汁后，从人血浆中分离的ACN及其代谢产物（PiACN）能够影响肿瘤细胞迁移，但不影响生长相关参数。这种抗迁移作用不仅取决于采血时间（果汁摄入后60分钟或14天），而且还取决于个体受试者。目前完全不清楚哪些化合物负责这些作用，以及它们通过何种机制诱导这些作用。该提案的目的是研究在使用富含ACN的果汁进行为期4周的干预研究后产生了哪些ACN和代谢物，以及它们是否能够诱导抗癌作用。将通过固相萃取法制备干预阶段前后所有受试者的PiACN，并用于体外功能研究。将研究胰腺（Panc-1、Asp-1）和结肠（HT-29、Caco-2）癌细胞系以及内皮细胞（HUVEC）的β-整合素、选择素和免疫球蛋白超家族成员的表达水平。相关信号传导途径（FAK-SFK-和氧化还原敏感性NF-κ B相关信号传导途径）的参与也是令人感兴趣的。除了PiACN对这些细胞本身的作用外，还将在体外研究其与细胞相关化合物的作用，以确定可能的相互作用。分析血液和粪便样本中的ACN及其代谢物，以及肠道微生物群的组成，应提供有关功能和代谢之间相互作用的进一步信息。

With about 20-30%, cancer is the second leading cause of death in Germany. The aggressiveness of a tumor depends on its metastasizing potential, which is characterized by the steps of invasion, intravasation and extravasation. New pharmacological therapeutics particularly aim at inhibiting cell adhesion molecules of tumor cells, e.g. integrins, selectins, and members of the immunoglobulin superfamily whose expression and pattern is strongly associated with the metastatic potential. They aim at the inhibition of tumor cell extravasation into the tissues and the formation of metastases. Nutritive factors that are able to influence these processes are currently being intensively investigated. Primarily, anthocyanins (ACN) belonging to the secondary plant ingredients are of particular importance. Their bioavailability is low, and they reach the lower gastrointestinal tract largely unchanged. Here, microbial fermentation products may be generated which could induce anti-carcinogenic effects after being absorbed into the systemic circulation. Preliminary experiments showed that ACN fermented by bacteria typically present in the human intestinal microbiota such as E. coli and Haf. alvei significantly reduced the migration of colorectal carcinoma and pancreatic carcinoma cells in vitro. Similarly, after the intake of an ACN-rich juice, ACN and their metabolites isolated from human plasma (PiACN) were able to influence tumor cell migration, but not to affect growth-associated parameters. This anti-migrative effect was not only dependent on the time of blood collection (60 min or 14 days after juice intake), but also on the individual subject. It is completely unclear which compounds are responsible for these effects and by which mechanism they induce these effects. The aim of the proposal is to investigate which ACNs and metabolites were generated after a 4-week-intervention study with an ACN-rich juice and if they are able to induce an anti-cancerogenic effect. The PiACN derived from all subjects before and after the intervention phase will be prepared by solid phase extraction and used for the in vitro functional studies. Pancreatic (Panc-1, Asp-1) and colon (HT-29, Caco-2) cancer cell lines as well as endothelial cells (HUVEC) will be investigated regarding their expression levels of β integrins, selectins and members of the immunoglobulin superfamily. The involvement of relevant signaling pathways (FAK-SFK- and redox-sensitive NF-B-associated signaling pathway) are also of interest. Besides the effects of PiACN on these cells itself, its effect regarding with pharmacologically relevant compounds is to be investigated in vitro in order to identify possible interactions. The analysis of blood and fecal samples with regard to ACN and their metabolites, as well as the composition of the intestinal microbiota, should provide further information on the interactions between function and metabolism.