metavectum Recklinghausen Modern Moleculare Diagnostic in Oncology

Aims of Molecular Oncology Early Diagnosis of Tumors Detection of disseminated Tumor-Cells and Micrometastasis Deciding of therapeutic Aims und Therapy Resistance Monitoring of Therapy Individualisised Therapy

Tumorprogression by genetic Alterations

Dissemination/Metastasis

Cancer is caused by genetic Alteration cancer cells obtain unnormal features Loss of Apoptosis Unkontrolled Growth Signal Unresponsive against growth inhibitors Metastasis creation Unlimited Replication Angiogenesis

Genes for Growth Factors Genetic Changes in Tumor-Cells Erb-b2, EGFR, c-Kit, PDGFR, K-ras Replications facilitating Genes Telomerase, c-myc Angiogenesis facilitating Genes VEGF Apoptosis facilitating Genes Bcl-2, p53

Circulating Tumor-cells (CTC) Risk-Factor for recurrance and metastasis. indicate risk for micro-metastasis. Easier indentification of patients profiting from adjuvant therapy. Suggested further reading: 1.Cristofanilli M. et al. Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med Aug 19;351(8): Allard WJ et al.: Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases Clin Cancer Res Oct 15;10(20):

Molekular Charakterisation of CTCs Isolation of mRNA from CTCs Measuring different Gene-Expression Quantitative real-time PCR of molekular Tumor-Marker (e.g.): Cytokeratine PSA (Prostata), G250 (kidney), AFP (liver) Growth / Cell-cycle-Regulating Gene (c-myc, erbb2, Telomerase)

Effect of Soy on Gene-Expression disseminated Tumor-Cells Five Patients per group suffering from metastastastic Prostate-, Ovary- and Breast -Ca received Chemo + fermented soy Clinical response in all patients Question: Can molecular factors predict clinical outcome of Ca- Therapy by fermented Soy ? Clincial : Bestimmung der modulierende Wirkung von Soja auf die Gen-Expression vor und nach 100 Tagen Einnahme des Soja- Extraktes

ER-beta ER-alpha DNA-Doppelstrang Funktion der Östrogenrezeptoren (ER) in der Zelle Zellteilungsaktivität wird gesteigert (Wucherung) Zellteilungsaktivität wird verringert a ß ERE a ß

Normale Zellen Tumorzellen mit Soja-Extrakt ER-beta ER-alpha ER-beta ER-alpha Expression der Östrogenrezeptoren (ER) alpha und beta ß a ß ß a a konstantes Verhältnis Absinken Anstieg Absinken Anstieg unbehandelt Brust / Ovar Prostata

Estrogen-receptor beta is induced by Haelan 951

Modulation of estrogen-receptor expression by Haelan 951 Circulating Tumor Cells (CTCs) of prostate cancer patients were analysed for expression of ER-alpha and ER-beta -> Expression of ER-beta was significantly induced by Haelan 951

relative Genexpression Mamma-Ca. patients n= 5 all shown breast cancer patients are ER-beta positive + Estrogenreceptor ER-beta Modulation of estrogen-receptor expression by Haelan 951

Estrogenreceptor-beta (ER-beta) Expression Relative Genexpression Mamma-Ca. Patients with Haelan951 = No Haelan951 = Haelan951 application Healthy persones n=7 Mamma-Ca. patients n=5 33,9 6,2 Healthy persones without Haelan951

Estrogen-receptor beta is induced by Haelan 951

Modulation of estrogen-receptor expression by Haelan 951 Circulating Tumor Cells (CTCs) of ovarian cancer patients were analysed for expression of ER-alpha and ER-beta -> Expression of ER-beta was significantly induced by Haelan 951

Stoffwechselweg der Apoptose c-myc mitogens proliferation apoptosis Bcl2 (survival signals) blocking NF-kB Bax Duales Signal von c-myc: Der apoptotische Stoffwechselweg wird blockiert durch bcl2 / bax und c-myc fördert Proliferation Nach Verlust des bcl2 Überlebenssignals, fördert c-myc die Apoptose

Apoptotic conditioning induced by Healan 951 CTCs of prostate cancer patients showed an increased gene expression ratio of apoptosis-triggering genes bax/bcl2 after Haelan951-therapy

without Haelan951 Haelan951 application 100 days relative Genexpression Mamma-Ca. patients n= 5 BAX/BCL2 no significant change 1,2 1,1 Apoptotic conditioning induced by Healan 951

Ovarian-Ca. patients n=6 Apoptotic conditioning induced by Healan 951

Apoptotic conditioning induced by Healan 951

The p53 Signaling Pathway Bax Bcl-2 p21 MDM2 p53 Caspases Apoptosis Cell Cycle Progression Cyclin D1 CDK4 Cyclin E CDK2 block Cell cycle inhibitor block (Ovarian Ca.)

M G1 p21 Involvement in Cell Cycle p53 p21 CIP1 Cyclin A/B CDK1 CDK2 M-phase entry block Cell cycle inhibitor Mitosis Proliferation DNA-Synthesis S Chromosomes Duplication G2

37,8 Cellcycle Inhibitor P21 is induced by Haelan951 without Haelan951 Haelan951 (100) Days) 1,9 0,5 0,9 1,0 0,6 relative Genexpression Prostata-Ca. patients n=5 cases 56,3 7,1 12,4 0,9

14,3 Cellcycle Inhibitor P21 is induced by Haelan951 without Haelan951 Haelan951 (100 days) 4,5 0,11 1,11 332,5 2,28 relative Genexpression Mamma-Ca. patients n=5 cases scale unit progressive 15,8 15,0 7,36 211,9

without Haelan951 Haelan951 application 100 days relative Genexpression Mamma-Ca. patients n= 5 p21 CIP increased 2,0 13,1 Cellcycle Inhibitor P21 is induced by Haelan951

Cellcycle Inhibitor P21 is induced by Haelan951

Schlussfolgerungen Alle Patienten zeigten auf die Gabe fermentierten Soja-Extraktes klinischen Response Fermentiertes Soja-Extrakt fördert den pro-apoptotischen Zustand der Prostata-Karzinomzellen durch Herunterregulation von Bcl2. Dies dürfte die Zellen sensitiver machen für eine Chemotherapie und somit für eine Kombinationstherapie mit konventionellen Chemotherapeutika sprechen. Zugabe von fermentiertem Soja induziert die Expression des antiproliferativen ER-beta bei Prostata- und Ovar-Ca Patienten. Fermentiertes Soja-Extrakt induziert die Transkription des p21, welches zu einem Stillstand des Zellzyklus führt und damit das Krebswachstum unterdrückt, sowohl bei Prostata-, Ovar- als auch bei Mamma-Ca Patienten.

The classical view of biological organization is to consider the flow of information from the genome to the transcriptome, to the proteome and then the metabolome. The metabolome can be used to monitor changes in the genome or to measure the effects of downregulation or upregulation of a specific gene transcript. metavectum Interaction

Invluence of Haelan951 to VEGF Expression without Haelan951 Haelan951 (100 Days) relative Genexpression Prostata-Ca. patients n=5 Mamma-Ca. patients n=5 Vascular Endothelial Growth Factor No alteration 0,97 1,46 37,5