1. Quercetin and Apoptosis within Cancer cells Lauren Walker Andrews University Dietetic Intern April 2015

2. Objectives: Define cellular apoptosis Define quercetin List quercetin rich food sources Explain how quercetin initiates apoptosis

3. Apoptosis Pre-programmed cell death ▫Cell shrinkage ▫Nuclear condensation ▫Nuclear fragmentation ▫Loss of adhesion to extracellular matrix

4. What is quercetin? Quercetin Flavonoid Anti- inflammatory Anti-viralAnti-tumor

5. Hypothesis: Can dietary quercetin initiate apoptosis within cancer cells? ?

6. Additional Questions: Dose of quercetin? Side effects of increased quercetin?

7. Forms of Quercetin: Quercetin tablets Quercetin powder Emergen-C Quercetin found in food

8. Quercetin-containing Foods Onion41.9 mg Asparagus23.6 mg Romaine lettuce12.0 mg Green pepper9.9 mg Cherry tomato3.3 mg Fuji Apple2.3 mg Broccoli1.6 mg Green Tea (100 mL)2.1 mg 100 milligrams = 330 micromoles

9. Comparing quercetin content (mg)

10. Zhaohui Study (2013) What?  The effect of quercetin among cell proliferation Hypothesis  quercetin would be able to induce apoptosis of human colon carcinoma cells (HT- 29 cells) through: ▫ capsases, ▫Bcl-2 family members, ▫and mitochondrial potential in HT-29 cells.

11. Zhaohui Study (2013) HT-29 Human colonic epithelial cells Incubated with 0, 60, 90, 120 micromoles of quercetin for 24 hours. Quercetin at 120 micromoles most effective (p<0.01) Results  Quercetin induces the activation of capsase 9 and capsase 3, which then stimulates the apoptosis signals.

12. Dose-dependent apoptosis Zhaohui L, Quanxin G. Induction of apoptosis in HT-29 cells by quercetin through mitochondrial-mediated apoptotic pathway. Animal Cells and Systems. 2013. (17) No.3 147-153

13. Senping et al. Study (2010) Focuses on Mcl-1 and Bax Human leukemia cells (U937 and HL-60) In Vivo ▫Reduced tumor size in quercetin exposed mice In Vitro

14. Senping et al Study (2010) In Vitro Results: Downregulation of Mcl-1 Activation of Bax Dose dependent Senping C, Ning G, Zhuo Z, et al. Quercetin Induces Tumor-Selective Apoptosis through Downregulation of Mcl-1 and Activation of Bax. Clinical Cancer Research. 2010;16:5679-5691

15. Senping et al Study (2010) Time dependent Senping C, Ning G, Zhuo Z, et al. Quercetin Induces Tumor-Selective Apoptosis through Downregulation of Mcl-1 and Activation of Bax. Clinical Cancer Research. 2010;16:5679-5691

16. Hyeonji et al study. (2013) Human Glioblastoma Cells (U373MG cells) Mitochondrial Apoptosis Protective Autophagy

17. Hyeonji et al study. (2013) Hyeonji K et al. Quercetin Induces Mitochondrial Mediated Apoptosis and Protective Autophagy in Human Glioblastoma U373MG Cells. Oxidative Medicine and Cellular Longevity. 2013. Article ID 596496. 10 pages

18. Hyeonji et al study (2013) Sub G-1 phase cells TreatmentSub G-1 Phase Control2.21 + 1.1 Quercetin Alone1.45 + 0.21 Chloroquine Alone5.18 + 0.96 Chloroquine + Quercetin18.68 + 3.02 Hyeonji K et al. Quercetin Induces Mitochondrial Mediated Apoptosis and Protective Autophagy in Human Glioblastoma U373MG Cells. Oxidative Medicine and Cellular Longevity. 2013. Article ID 596496. 10 pages

19. Hyeonji et al study. (2013) Pre-treatment of cells with chloroquine Hyeonji K et al. Quercetin Induces Mitochondrial Mediated Apoptosis and Protective Autophagy in Human Glioblastoma U373MG Cells. Oxidative Medicine and Cellular Longevity. 2013. Article ID 596496. 10 pages

20. Hyeonji Kim et al study. (2013) Quercetin combined with an autophagy inhibitor may be an excellent therapeutic approach. ▫First de-sensitize ▫Then attack!

21. Summary Quercetin has been shown to induce apoptosis within different types of cancer cells. ▫Capsase-9, Capsase-3 ▫Bcl-2 family members (Mcl-1, Bax, Bcl-2) ▫mitochondrial potential in HT-29 cells. Difficult to control the exact plasma concentration of quercetin, unsure how to dose it for individuals. Further research is needed on dose and different lines of cancer cells.

22. Questions?

23. Bibliography Zhaohui L, Quanxin G. Induction of apoptosis in HT-29 cells by quercetin through mitochondrial- mediated apoptotic pathway. Animal Cells and Systems. 2013. (17) No.3 147-153 Wong R. Apoptosis in cancer: from pathogenesis to treatment. Journal of Experimental and Clinical cancer research. 2011, 30:87 Zhaohui L, Quanxin G. Induction of apoptosis in HT-29 cells by quercetin through mitochondrial- mediated apoptotic pathway. Animal Cells and Systems. 2013. (17) No.3 147-153 Haruno et al. Estimated Daily Intake and Seasonal Food Sources of Quercetin in Japan. Nutrients. 2015. 7, 2345-2358. Walle T, Walle UK, Halushka PV. Carbon Dioxide is the major metabolite of quercetin in humans. Journal of Nutrition. 2001. (10):2648-52 Senping C, Ning G, Zhuo Z, et al. Quercetin Induces Tumor-Selective Apoptosis through Downregulation of Mcl-1 and Activation of Bax. Clinical Cancer Research. 2010;16:5679-5691 Hyeonji K et al. Quercetin Induces Mitochondrial Mediated Apoptosis and Protective Autophagy in Human Glioblastoma U373MG Cells. Oxidative Medicine and Cellular Longevity. 2013. Article ID 596496. 10 pages Ouyang L et al. Programmed cell death pathways in cancer: a review of apoptosis, autophagy, and programmed necrosis. Cell Proliferation. 2012 (45) 487-498 Guomin Niu et al. Quercetin induces apoptosis by activating caspase-3 and regulating Bcl-2 and cyclooxygenase-2 pathways in human HL-60 cells. Biochemistry and Cell Biology. 2011. (43) 30-37