Lycopene and Soy Isoflavones in

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					Lycopene and Soy Isoflavones in
      Cancer Prevention
         Omer Kucuk, MD
        Professor of Medicine
       Winship Cancer Institute
          Emory University
       Atlanta, Georgia, USA
NUTRITION AND
   CANCER
Let food be your
medicine.
          -Hippocrates
               400 BC
Tomato consumption and cancer
 72 studies identified (observational
  studies)
 57 reported inverse association
  between tomato intake or blood
  lycopene level and the risk of cancer
  (at a definitive anatomic site)
   35 of these inverse associations were
    statistically significant
 No study indicated a positive
  association
                                 Giovannucci E., 1999
         Lycopene reduces the risk of various types of cancer
                       Statistically Significant   Not Statistically Significant

               Total
       Oral, Larynx
        Esophagus
           Stomach
              Colon
            Rectum
          Pancreas
               Lung
            Prostate
            Bladder
              Breast
             Cervix
              Ovary
      Mesothelioma

Giovanucci, JNCI   RR:            0.5              1          1.5
91, 317-331 1999
Convincing protective effect

   lung, stomach, prostate



Suggestive protective effect

  colon-rectum, pancreas, breast, cervix,
  oral cavity, esophagus
              The composition of tomato

water          94    %   vitamin C     160-240 mg/kg
protein        0.7   %   vitamin E     8 – 12    mg/kg
fat            0.3   %
                         lycopene      15 - 56 mg/kg
carbohydrat    3.1   %
fibre          1.2   %   carotenoids   6 - 8     mg/kg

                         flavonoids    5 - 50    mg/kg

                         phenols       10 - 50 mg/kg
                         manganese     1 – 1.5   mg/kg

                         zinc          1 – 2.4   mg/kg

                         iron          4 - 5     mg/kg

                         copper        0.1-0.9 mg/kg
                         folates       150-390 g/kg
   Effect of tomato product intake on
          prostate cancer risk
                    1.10
                                                      tomato juice
                    1.00
    Relative risk



                    0.90
                                                      tomato fruits
                    0.80
                                                      pizza
                    0.70
                                                      tomato sauce
                    0.60
                           0      1       2   3
                               Servings
Adapted from Giovanucci et al. JNCI 87, 1767 (1995)
Carotenoid intake and prostate cancer

                       1.10

                       1.00                               b-carotene
       Relative risk


                       0.90

                       0.80                               lycopene
                       0.70

                       0.60
                              0    1    2   3    4    5
                                  Carotenoid intake
Giovanucci et al. JNCI 87, 1767 (1995)
 Steps in carotenoid biosynthesis
b-carotene


Lycopene

Neurosporene

-carotene

Phytofluene

 Phytoene
   Effect of tomato extract supplementation on IGF-I and
  lycopene levels in colon cancer patients (Sharoni, Levy)

                    Baseline       After supplementation

              A. IGF-I                     B. Plasma lycopene
        210   P< 0.001    N.S.          0.6 P< 0.05     N.S.
        175
ng/ml




        140                  -6         0.4


                                   µM
                             %
        105
                  -29
        70         %                    0.2
        35

         0                               0
              lycopene   placebo              lycopene   placebo
                n=21     n=18                   n=21      n=18
Astaxanthin and lycopene inhibit cancer cell proliferation
                                            Mammary cancer                          Prostate cancer

                                   MCF-7              MDA-231              LNCaP                 DU-145
                                                               Astaxanthin
                                                               Lycopene
cell growth (percent of control)




                                   120                120                 120                   120

                                   100                100                 100                   100

                                   80                 80                  80                    80

                                   60                 60                  60                    60

                                   40                 40                  40                    40

                                   20                 20                  20                    20

                                    0                  0                   0                     0
                                        0    5   10        0   5     10         0    5    10         0   5   10


                                                                   Carotenoid [M]
Androgen- and IGF-I-stimulated growth in LNCaP is
  inhibited by astaxanthin and lycopene (Sharoni)

                                                                 Control
  [3H]thymidine incorporation (cpm)




                                      1600                       Astaxanthin
                                                                 Lycopene
                                      1200



                                      800



                                      400



                                        0

                                             No addition   DHT      IGF-I
Lycopene and astaxanthin reduce PSA level in
    prostate cancer cells (Sharoni, Levy)
                      0h          6h       12 h        24 h       36 h



                              1   2    3 1 2 3     1     2 3 1 2 3
                      16                                       Testosterone (1)
    Arbitrary units



                      12
                                                               Tes+ astaxanthin (3)
                      8


                      4
                                                               Tes+lycopene (2)


                      0
                                      12      24          36
                           Time after testosterone (hour)
The synergistic effect at low concentration of
 carotenoids in LNCaP prostate cancer cells


        Thymidine incorporation (cpm)
                                        1500



                                                                Phytoene
                                        1000

                                                                Lycopene

                                         500
                                                    +Lycopene
                                                     0.2 M


                                           0
                                               0    1   2   3   4

                                                   carotenoid [µM]
                          Tomato carotenoids activate expression from
                             nuclear receptor RE (Sharoni, Levy)
                        100
                         30
                              atRA                                   RAR Response
Reporter activity (%)


                         20                                          Element (DR-5)

                         10

                          0
                        100
                              Cigl.                                  PPARg Response
                        80
                                                                     Element (DR-1)
                        60
                        40
                        20
                         0
                              agonist lycopene phytoene b-carotene
Pilot study of lycopene in patients
        with prostate cancer
MEN SCHEDULED FOR RADICAL PROSTATECTOMY
                    |
           BIOPSY AND BLOOD SAMPLES
                    |
 R    A   N    D    O    M    I     Z    E
     |                                |
LYCOPENE* 30 MG/DAY            NO LYCOPENE
    |                                  |
 PROSTATECTOMY AND BLOOD SAMPLES AFTER 3
            WEEKS OF INTERVENTION

                                  *Lyc-O-Mato
    Carotenoid content of
    Lyc-O-Mato® capsule
All-E(trans)-Lycopene   13.50 mg
5-Z(cis)-Lycopene        1.05 mg
5Z,5'Z-Lycopene          0.45 mg
b-Carotene               0.16 mg
g-Carotene               0.09 mg
-Carotene               0.24 mg
Phytofluene              1.03 mg
Phytoene                 1.16 mg
   Typical carotenoid profile of a prostate
specimen after 3 weeks on 30 mg/d Lycomato
 Pathologic Tumor Stage
                                LYCOPENE              CONTROL
                                GROUP                 GROUP
                           (N=15)            (N=11)       p
---------------------------------------------------------------
Confined to Prostate                   11                 2
Not Confined to Prostate*               4                 9          0.02
-----------------------------------------------------------------------------
*Resection margins are positive or extra-prostatic invasion is
    present.
 Prostatic Tumor Volume
                 LYCOPENE          CONTROL
                 GROUP             GROUP
                  (N=15)            (N=11)               p
---------------------------------------------------------------
Volume 4cc< 12                         5
Volume >4cc           3                6               0.22
             Plasma PSA Level
                (mean ng/dl)
                                 LYCOPENE             CONTROL
                                 GROUP                GROUP
                                 (N=15)               (N=11)            p
-----------------------------------------------------------------------------

Pre-Intervention                6.89                  6.74
Post-Intervention               5.64                  7.65

                                 -18%                 +14%            0.22
---------------------------------------------------------------------------
      Extent of Prostatic
Intraepithelial Neoplasia (PIN)

                                LYCOPENE              CONTROL
                                GROUP                 GROUP
                                (N=15)                (N=11)                p

-----------------------------------------------------------------------------------

Focal                           5                     0
Multifocal/diffuse              10                    11                    0.05

-----------------------------------------------------------------------------------
          Expression of Biomarkers
            In Prostatic Tumor
                      Lycopene                        Control
                       (n=4)                          (n=4)                 p
------------------------------------------------------------------------------------
  Connexin-43          0.6281                         0.2514                0.13

  bcl-2                0.5430                         0.5056                0.59

  bax                  1.0531                         0.6848                0.33

  bax/bcl-2            1.9279                         1.4896                0.54

-------------------------------------------------------------------------------------
  Expression of Biomarkers
  in Benign Prostatic Tissue
                                Lycopene              Control
                                (n=8)                 (n=6)                 P
-------------------------------------------------------------------------------
Lycopene (ng/gm)                0.5284                0.3587                0.02
                                (n=5)                 (n=3)

Connexin 43                     0.6436                0.5101                0.44

bcl-2                           0.6324                0.5765                0.31

bax                             0.6218                0.7892                0.28

------------------------------------------------------------------------------------
             Conclusions
• Lycopene (Lycomato ®) supplementation
  for three weeks is well tolerated by patients
  with prostate cancer without any side
  effects.
• After three weeks of lycopene
  supplementation prostate tissue levels of
  lycopene increase associated with
  upregulation of gap junctional protein Cx43.
             Conclusions
• Clinical markers, including HGPIN and
  serum PSA level improve in patients
  receiving lycopene supplementation
• Although these results are preliminary other
  investigators have obtained similar results
  (Bowen et al, JNCI) suggesting activity of
  lycopene in patients with prostate cancer.
               Soy Isoflavones
                              CH3                  B. Genistein
A. Tamoxifen
               O-CH2-CH2-N
                              CH3                                     OH
                                                   HO             O



                                             HO
                                                                  O



                                                        OH
                                                  CH3
                         C. Estradiol (E2)




                       HO
 Soy isoflavones and cancer
• Epidemiologic studies show an inverse
  association between dietary soy intake and
  cancer risk (breast, prostate, lung, colon,
  head and neck and others)
• Genistein and daidzein are the most
  abundant isoflavones in soy
• Genistein has activity against a variety of
  cancer cells in culture, animal model and
  clinical studies
     Genistein and Prostate
            Cancer
• Inhibits growth and induces apoptosis in Pca cells
• Growth inhibition mediated by G2/M cell cycle arrest
  and up-regulation of p21WAF1
• Down-regulates cyclin B1, CDKs, Bcl-2/Bcl-xL
• Up-regulates Bax expression and induces translocation
  of Bax to Mitochondria
              Genistein
• Down-regulates MMP-2, MMP-9, uPA,
  c-IAP and VEGF
• Inactivates Akt and NF-kB (by
  inhibiting IKK)
  – blocks nuclear translocation of p50 and p65
  – inhibits phosphorylation of IkBa
  – decreases MEKK1 kinase activity
        Growth Factor, Cytokine               Genistein

             NIK        IKK            Akt
                                   P
                             p50       p65 NFkB

                   P
                                                    Active
       IkB                               p50 p65    NFkB
    Degradation
Ubiquitin-Proteasome


                                   Nuclear Translocation
                       p50 p65
                       kB site           •cIAP-1, XIAP, MMP-9,
                                         uPA, VEGF, etc.
                   Transcription
 Effect of Dietary Genistein on MMP Gene
  Expression in Experimental Metastasis




Affymetrix Human Genome U95 or U133A Array
      Cluster Analysis According to
           Biological Function
Numbers of altered genes in different categories in
PC3 bone tumors after genistein treatment
   Category                                  Up     Down
   apoptosis                                  12          1
   cell cycle arrest, negative regulation     13          0
   of cell proliferation and transcription
   signal transduction, chemotaxis            10          7
   regulation of transcription and            11          10
   protein biosynthesis
   oncogenesis                                 8          4
   Based on Cluster, Onto-Express, and GenMAPP Software
Effects of genistein on gene expression*




          *Based on in vitro and in vivo gene profiling with and without genistein
 Dietary Genistein and Experimental PC3
Bone Metastasis (Neoplasia 6:354-363, 2004)



                            p0.0001
                            p=0.0003




               control    prevention
         SCID-Human Model of Prostate
            Cancer Bone Metastasis




                                   1 cm




Normal        PC3          LNCaP          LUCaP 23.1
Phase II clinical trial of soy isoflavones in
      patients with prostate cancer
 Study subjects:
 • Histologically proven prostate cancer with
   PSA progression.
 • No other therapy for prostate cancer, except
   patients already on LHRH analogue were
   required to continue it.
 • Patients had to demonstrate a rising trend
   with three successive elevations at a
   minimum interval of two weeks or at least
   two PSA values at least 2 weeks apart with a
   minimum PSA of 10 ng/ml.
             Treatment
• Soy isoflavone (Novasoy®) 100mg orally
  twice daily.
• Treatment duration: Maximum 6 months.
• Treatment compliance: Medication
  calendars were completed by study
  subjects and remaining pill counts were
  done on returned bottles.
Phase II clinical trial of soy isoflavones in
      patients with prostate cancer

 • Group 1 (n=4)
    – previously untreated (watchful waiting)
 • Group 2 (n=18)
    – PSA-failure, post-surgery or -radiation
 • Group 3 (n=19)
    – PSA-failure, post-hormone therapy
      (hormone refractory)
            PSA response
• Group 1
 Stable 3/4 patients (75%)
• Group 2
 Stable 15/18 patients (83%)
• Group 3
 Stable 6/19 patients (32%)
                    Plot of predicted rise in log PSA with time
          6.5


          5.5
log PSA




                       G3
          4.5
                                   intervention

          3.5


          2.5             G2

          1.5
                0        100       200      300      400          500
                                         Days
   Toxicity and compliance
• There was no grade 2 or greater
  toxicity. No patients stopped treatment
  because of side effects.
• Compliance was excellent with all
  subjects taking more than 80% of
  prescribed study pills.
   Phase II clinical trial of lycopene and
    soy isoflavones in prostate cancer
• Lycopene and soy isoflavones have activity
  against prostate cancer in cell culture, animal and
  clinical studies
• We hypothesized that the combination of
  lycopene and soy isoflavones taken together may
  be more effective in patients with prostate cancer
• We conducted a clinical trial in patients with PSA
  relapse prostate cancer to test the hypothesis
         Patient eligibility
• Histologically proven prostate cancer with
  PSA progression.
• No other therapy for prostate cancer, except
  for the patients who were already on LHRH
  analogue were required to continue it.
• Patients had to demonstrate a rising trend
  with three successive elevations at a
  minimum interval of two weeks or at least
  two PSA values at least 2 weeks apart with a
  minimum PSA of 10ng/ml.
              Treatment
• Lycopene arm: Lycomato ® 15mg orally
  twice daily.
• Combination arm: Lycomato ® 15mg +
  soy isoflavone (Solgen®) 40mg orally
  twice daily.
• Treatment duration: Maximum 6 months.
• Treatment compliance: Patients were
  given a medication calendar and pill counts
  were done on returned bottles.
Phase II clinical trial of lycopene and soy isoflavones in prostate cancer



 Patient Characteristics      Lycopene N=38 (53.5%)     Lycopene + Soy Isoflavone N=33 (46.5%)



 Median Age                   73 years                  76 years
 Range                        (57-89 years)             (50-91 years)
 Race
 Caucasian                    24 (63%)                  23 (70%)
 African American             12 (31%)                  9 (27%)
 Other                        2 (6%)                    1 (3%)
 Prior Systemic Therapy
 Hormones                     14 (36%)                  11(33%)
 None                         24 (64%)                  22 (67%)
 Presence of metastases
 Present                      8 (21%)                   10 (30%)
 Absent                       30(79%)                   23 (70%)
 Median PSA (Range)           6.1ng/ml (1.1-147ng/ml)   6.9ng/ml (0.8-60.9ng/ml)




                           Vaishampayan U, et al. Nutr Cancer 2007.
               Results

• 35 (95%) of 37 patients on lycopene
  had PSA stabilization
• 22 (67%) of 33 patients on lycopene +
  soy had PSA stabilization.
• In both treatment arms, there was a
  decline in the rate of PSA rise in
  hormone sensitive (p=0.015) and
  hormone refractory (p=0.017) patients.
Phase II clinical trial of lycopene and soy isoflavones in patients with prostate ca.


                                        PSA rise before and after treatment
                                                  Hormone Sensitive Patients
                   7


                   6
   Predicted PSA




                   5


                   4


                   3


                   2

                       -400         -300         -200           -100             0          100   200
                                                                Days
                        Solid line is lycopene only arm, and dashed line is lycopene+soy arm
Phase II clinical trial of lycopene and soy isoflavones in patients with prostate ca.


                                         PSA rise before and after treatment
                                                 Hormone Refractory Patients


                      40



                      30
      Predicted PSA




                      20



                      10



                      0
                           -200           -100               0                100           200
                                                          Days
                       Solid line is lycopene only arm, and dashed line is lycopene+soy arm
             Conclusions
• The results of this phase II randomized
  clinical trial demonstrate the role of
  lycopene in decelerating the rate of PSA rise
  in patients with relapsed prostate cancer.
• The results suggest that soy isoflavones in
  combination with lycopene may have an
  adverse effect on the PSA response to
  lycopene.
     Lycopene, Soy and Zinc
     Prostate Cancer Studies
•   Wayne State University, Karmanos Cancer Institute
     –   O. Kucuk, M. Cher, E. Pontes
     –   W. Sakr
     –   F. Sarkar, Y. Li,
     –   A. Prasad
     –   Z. Djuric
     –   M. Banerjee
     –   G. Hillman, J. Forman
•   Michael Pollak (McGill University, Montreal)
•   Fred Khachik (University of Maryland)
•   M. Baykara, A. Ciftcioglu, G. Yildirim-Kupesiz, F. Koseoglu-Andic
    (Akdeniz University, Turkey)
•   F. Saatcioglu, B. Lazarevic (Oslo University, Norway)
•   H. Mukhtar (University of Wisconsin, Madison, WI)
•   D. Doerge (National Center for Toxicology, Jefferson, AK)
Let food be your
   medicine.
Hippocrates, 400 BC

				
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