Enhanced Chemopreventive Effect by Combining Quercetin and Green tea in Prostate Cancer

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1 Enhnced Chemopreventive Effect y Comining Quercetin nd Green te in Prostte Cncer Piwen Wng, MD, PhD Assistnt Professor, Division of Cncer Reserch nd Trining Chrles R. Drew University of Medicine nd Science

2 (Cncer Fcts & Figures 2015)

3 (Cncer Fcts & Figures 2015)

4 Age-Stndrdized Prostte Cncer Incidence nd Mortlity Rtes y World Are. Source: GLOBOCAN (Jeml A, et l. CA CANCER J CLIN 2011;61:69 90)

5 Prostte cncer Prostte cncer: Science nd clinicl prctice. Edited y: J Mydlo & C Godec. 2013)

6 Te From the plnt Cmelli sinensis Green te (GT): - Not fermented - Polyphenol monomer (EC, EGC,ECG, EGCG) Oolong te: - Prtilly fermented Blck te: - Fully fermented - Dimers nd polymers (theflvins, theruigins) (Alipoor, et l. Asin J Clin Nutr. 2012, 4 (1): 1-15

7 GT nd GT polyphenols Anti-crcinogenesis - Skin, lung, orl cvity, esophgus, stomch, intestine, colon, liver, pncres, ldder, mmmry glnd, prostte Dietes nd oesity Crdiovsculr diseses Neurodegenertive diseses Osteoporosis

8 Possile trgets for the cncer preventive ctivity of (-)-epiglloctechin-3-gllte (EGCG). (Yng CS, et l. Nture Reviews Cncer 2009, 9, )

9 Pooled odds rtio (OR) of GT consumption (highest versus non/lowest) nd risk of prostte cncer (Zheng J. et l Nutr nd Cncer 63(5): )

10 Intervention studies: GT nd prostte cncer Te, Country Effect Reference GT extrct, Itly, 600 mg GT extrct, USA, 800 mg EGCG (single rm) GT extrct, USA, 800 mg EGCG (plceo controlled) Reduced progression from HG-PIN to prostte cncer: Control 30% GTE 3% Decresed serum PSA, HGF, VEGF in prostte cncer ptients A decresing trend of PSA, in prostte cncer ptients Bettuzzi S et l McLrty J et l Nguyen MM et l. 2012

11 A phse II clinicl tril of te in prostte cncer Screening Rndomize of eligile sujects Plceo (N=33) GT (N=34) BT (N=26) 3-8 weeks intervention Urine polyphenol metolites 8-OHdG Prostte polyphenol metolites Serum polyphenol metolites ex vivo iossy PSA Prostte prolifertion poptosis ROS Bx/Bcl-2 NFkB

12 (Henning. et l Prostte 75: )

13 (Henning. et l Prostte 75: )

14 (Henning. et l Prostte 75: )

15 Limittions of GT in chemoprevention Low iovilility of GT polyphenols Extensively methyltion in vivo EGCG Concentrtion (pmol/g tissue) EGCG 4"-MeEGCG ECG 120 CH 3 Ctechol-O-methyltrnsferse (COMT) Cell viility (% of control) c NT 4"-MeEGCG 40uM EGCG 40uM 4"-MeEGCG 80uM EGCG 80uM cd cd 0 0h 24h 48h 72h (Wng P, et l Cncer Prev Res 3(8):985-93)

16 Quercetin (Q) A flvonoid commonly found in fruits nd vegetles - red onions, pples, roccoli, erries, nd te A nturl inhiitor of ctechol-o-methyltrnsferse (COMT) nd multidrug resistnce-relted proteins (MRPs)

17 Quercetin in comintion with EGCG in vitro cellulr sorption Concentrtion (pmol/mg cytosolic protein) EGCG 80uM EGCG 4"-MeEGCG Quercetin Isorhmnetin c d c Quer 5uM Quer 10uM EGCG + Quer 5uM EGCG + Quer 10uM d c LNCP d c d Concentrtion (pmol/mg cytosolic protein) EGCG 80uM EGCG 4"-MeEGCG Quercetin Isorhmnetin c d c Quer 10uM Quer 20uM EGCG + Quer 10uM EGCG + Quer 20uM d PC-3 c d c d The groups with the sme superscript letter represent significnt difference etween groups (P<0.05) (Wng P, et l Nutr Cncer 64(4): )

18 Quercetin in comintion with EGCG in vitro Antiprolifertive effect 110 LNCP 110 PC Cell viility (% of control) NT EGCG 40uM Quer 5uM Quer 10uM EGCG 40um + Quer 5uM EGCG 40um + Quer 10uM cd cd c cd Cell viility (% of control) c c NT EGCG 40uM Quer 10uM Quer 20uM EGCG 40um + Quer 10uM EGCG 40um + Quer 20uM cd cd Time (Hours) Time (Hours) Compred to, control (NT);, EGCG; c, Q 5umol/L; d, Q 10umol/L, p<0.05 (Wng P, et l Nutr Cncer 64(4): )

19 Quercetin in comintion with EGCG in vitro COMT inhiition Vnillic cid formed (nmol/h/mg protein) NT EGCG 40uM Quer 10uM Quer 20uM EGCG 40uM + Quer 10uM EGCG 40uM + Quer 20uM c c d e Time (Hours) d e c d e f 24h 48h COMT concentrtion (reltive to NT) NT EGCG 40 μm Quer 10 μm NT EGCG 40uM Quer 10uM EGCG 40uM + Quer 10uM c Time (Hours) EGCG 40 μm+ Quer 10 μm c COMT β-ctin COMT β-ctin (Wng P, et l Nutr Cncer 64(4): )

20 Incresed nti-prolifertive effect y EGCG + Q in lung, liver nd kidney cncer cells Cell viility (% of control) NT EGCG 40uM Quer 10uM Quer 20uM EGCG 40um + Quer 10uM EGCG 40um + Quer 20uM 0h 24h 48h 786-O (kidney cncer) cd cd A549 (lung cncer) c cd cd Cell viility (% of control) cd cd NT EGCG 40uM Quer 10uM Quer 20uM EGCG 40um + Quer 10uM EGCG 40um + Quer 20uM Hep G2 (liver cncer) 0h 24h 48h c cd 100 Cell viility (% of control) NT EGCG 40uM Quer 10uM Quer 20uM EGCG 40um + Quer 10uM EGCG 40um + Quer 20uM c cd cd cd h 24h 48h (Wng P, et l Food Funct.3(6):635-42)

21 Xenogrft SCID mouse model 0.5 million LAPC4 cells Weeks When tumor ws plple - strt of intervention

22 Quercetin in comintion with GT in SCID mice GT (Celestil Sesonings, Boulder, CO): 1 te g rewed in 240 ml oiling wter for 5min; 0.07% GT polyphenols Concentrtion (mg/l wter) Q (Sigm-Aldrich): 0.4% in diet AIN 93G+Quercetin 0 EGC EGCG EC ECG

23 Xenogrft tumor growth 1.2 Tumor volume (cm 3 ) Tumor inocultion Tretment strted Control GT Q GT + Q c (16%) (21%) c (45%) Time (Weeks) Compred to, control group;, Q group; c, GT group, p<0.05 (Wng P, et l J Nutr Biochem 25(1):73-80)

24 Food nd wter consumption nd ody weight Dily consumption of food (g) nd wter (ml) per mouse Control GT Q GT+Q Food Wter mouse weight (g) Control GT Q GT+Q Weeks

25 GT+Q incresed the iovilility nd decresed methyltion of GT polyphenols in tumor tissues Concentrtion (pmol/g tissue) EGCG 4"-MeEGCG EGC ECG EC quercetin isorhmnetin GT Q GT+Q (Wng P, et l J Nutr Biochem 25(1):73-80)

26 GT+Q incresed the iovilility nd decresed methyltion of GTPs in other tissues Concentrtion (pmol/g tissue) EGCG 4"-MeEGCG EGC ECG EC quercetin isorhmnetin Lung GT Q GT+Q compred to individul tretment, p<0.05 Concentrtion (pmol/g tissue) Concentrtion (pmol/g tissue) 450 EGCG 4"-MeEGCG EGC ECG EC 400 quercetin isorhmnetin Kidney GT Q GT+Q EGCG 4"-MeEGCG EGC ECG EC quercetin isorhmnetin Liver GT Q GT+Q (Wng P, et l Food & Function 3: )

27 Modultion of protein mrker expression COMT DNMT AR PSA VEGF GAPDH Control GT Quercetin GT+Q Protein concentrtion (reltive to control) Control GT Q GT+Q 0.0 COMT DNMT AR PSA VEGF (Wng P, et l J Nutr Biochem 25(1):73-80)

28 Inhiition of prolifertion mrker-ki67 (Wng P, et l J Nutr Biochem 25(1):73-80)

29 GT+Q in comintion with drugs for chemotherpy of dvnced prostte cncer

30 GT+Q+Docetxel: Enhnced inhiition of PC-3 tumor growth 1.6 Tumor volume (cm 3 ) Tumor inocultion Control GT+0.4%Q Doc 5 GT+0.4%Q+Doc 5 Doc 10 Tretment strted Lst Doc injection c c c (100%) (38%) (34%) Weeks Dt re presented s men±se. Different letters t ech time point indicte sttisticlly significnt difference etween groups, p<0.05 (mnuscript sumitted)

31 Summry nd Conclusion Quercetin in comintion with GT significntly enhnces the nti-crcinogenic effect in vitro nd in vivo The comined effect ws ssocited with n incresed iovilility nd decresed methyltion of GT polyphenols These results wrrnt future studies to confirm the synergistic effect of the comintion of GT nd quercetin in humns In ddition, this comintion my improve clinicl ppliction in tretment of dvnced prostte cncer y sensitizing tumor cells to chemotherpy

32 Acknowledgement Dr. Jy Vdgm (Chrles R. Drew University) Drs. Susnne Henning, Dvid Heer (UCLA Center for Humn Nutrition), Jonthn W. Sid, Clr E. Mgyr, Ngn Don (UCLA Deprtment of Pthology) Grnt supports from the Ntionl Institutes of Helth [NIH, NCI, NIMHD, NCATS; Grnt Nos. U54 CA , U54MD007598, UL1TR (J.V. Vdgm)], nd NIH/Ntionl Center for Advncing Trnsltionl Sciences UCLA CTSI [Grnt No. KL2TR (P. Wng)]

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