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

癌症是德国第二大死因，约占20-30%。肿瘤的侵袭性取决于其转移潜力，其特点是浸润、内渗和外渗。新的药物治疗方法特别针对抑制肿瘤细胞的细胞粘附分子，如整合素、选择素和免疫球蛋白超家族成员，其表达和模式与转移潜力密切相关。它们的目的是抑制肿瘤细胞向组织外渗和转移的形成。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）属于次生植物成分，具有特别重要的意义。它们的生物利用度很低，到达下胃肠道时基本不变。在此过程中，微生物发酵产物被吸收进入体循环后可能产生抗癌作用。初步实验表明，ACN由大肠杆菌和Haf等人类肠道微生物群中常见的细菌发酵而成。肺泡在体外可明显减少结直肠癌和胰腺癌细胞的迁移。同样，在摄入富含ACN的果汁后，从人血浆中分离出的ACN及其代谢物（PiACN）能够影响肿瘤细胞的迁移，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），而且取决于个体受试者。目前还完全不清楚是哪些化合物造成了这些影响，以及它们通过何种机制引起了这些影响。该提案的目的是研究在使用富含acn的果汁进行为期四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。所有受试者干预前后的PiACN将采用固相萃取法制备，用于体外功能研究。胰腺癌（Panc-1, Asp-1）和结肠癌（HT-29, Caco-2）细胞系以及内皮细胞（HUVEC）将被研究其β整合素，选择素和免疫球蛋白超家族成员的表达水平。相关信号通路（FAK-SFK-和氧化还原敏感NF-b相关信号通路）的参与也令人感兴趣。除了PiACN对这些细胞本身的作用外，其与药理学相关化合物的作用还有待于在体外研究，以确定可能的相互作用。对血液和粪便样本进行ACN及其代谢物的分析，以及肠道微生物群的组成，应能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.