NORMAL AND CANCER CELL GROWTH REGULATION

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NORMAL AND CANCER CELL GROWTH REGULATION
Normal Cells
Tumor Cells
CANCER CELLS
SAME AS TUMOR CELLS BUT 1) APOPTOTIC PROGRAM IS
READY AND REMAIN INHIBITED 2)APOPTOSIS IS
INDUCABLE
24hrs
24hrs
24hrs
24hrs
24hrs
Not Programmed for Cell Death (Non Apoptotic)
Undifferentiated Dividing Cells
Differentiated Cells
n1
2n
APOPTOTIC (Programmed) CELL DEATH
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Structure of Apoptotic Drugs
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Apoptotic Colo-205 by Drugs
Cis-Platin 150 µM 48 hrs
L-PPMP 15 µM 48 hrs
Control Cells were synchronized with 0.5 mM
HU(24 hrs) X 2
Melphalan 16 µM 48 hrs
Tamoxifen 16 µM 48 hrs
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IDENTIFICATION OF APOPTOTIC CANCER CELLS USING
FLUORESCENT DYES
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APOPTOSIS OF BREAST CANCER CARCINOMA (SKBR3)
CELLS(EFFECT OF Betulinic Acid/2hrs)
Control Phase Contrast
Control Blue with PSS1
Control Red with PPI
10uM BetA Phase Contrast
10uM BetA Blue with PSS1
10uM BetA Red with PPI
40uM BetA Phase Contrast
40uM BetA Blue with PSS1
40uM BetA Red with PPI
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APOPTOSIS OF BREAST CANCER CARCINOMA (SKBR3)
CELLS(EFFECT OF GD3/6hrs)
Control Phase Contrast
Control Blue with PSS1
Control Red with PPI
10uM GD3 Phase Contrast
10uM GD3 Blue with PSS1
10uM GD3 Red with PPI
80uM GD3 Phase Contrast
80uM GD3 Blue with PSS1
80uM GD3 Red with PPI
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SUBSTRATES USED FOR CGASE ASSAYS
GgOse43-TCer Galb1-3GalNAcb1-4Galb1-4Gl
c-3-TCER nLcOse54,5-TCer
Gala1-3Galb1-4GlcNAcb1-3Galb1-4Glc-4,5-TCER
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TARGETED DELIVERY OF DRUGS TO CANCER CELLS (By
LIPOSOME BULLETS)
Mab (CSLEX)
SA-Lex
Synthetic Liposomes Containing Anti-cancer
Drugs/ cis-Platin, Tamoxifan, Melphalan,
Betulinic Acid L/D-PPMP,
L/D-PDMP, GD3, Ceramides, GD1b, GT1b
Metastatic Cancer Cells (Colon, Breast,Neuronal,
Prostate, ALL)
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ACKNOWLEDGEMENTS

• At Notre Dame Outside Collaboration
• Dr. Manju Basu 1.Dr. Jin-ichi Inokuchi
• Brian Mikkula (Hokkaido University)
• Maria Miguel 2.Dr. Sipra Banerjee
• Matthew Bradley (Cleveland Clinic)
• Rui Ma
• Patrick Boyle
• Chris Moulton
• Shannon Bradley

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Summary

• Human colon carcinoma (Colo-205) cells were
  synchronized in the presence of 0.5 mM
  hydroxyurea (18hrs x 2).
• Synchronized carcinoma cells showed apoptosis,
  programmed cell death, in the presence of either
  L/D-PDMP or -PPMP (0.5 to 8.0 mM). cis-platin (25
  150 mM) .
• Agarose gel electrophoresis showed the
  characteristic DNA laddering of the apoptotic
  process.
• Apoptotic cells produced by cis-platin showed
  lower activities of Caspases-1 . However,
  Caspase-3 is not affected and perhaps Casoase-3
  is involved in the degradation of the chromosomal
  DNA.
• Apoptotic cells produced by PDMP PPMP (2.0-
  4.0.0 mM) in the presence of L-14C/or 3H-Serine
  (24-48 hrs) showed increased biosynthisis of
  CERAMIDE and at higher concentration decreased
  both Glc-cCer and highier Oligp-glycolipids.
• Binding of cis-platin to Zn-binding domain of
  DNA-polymerase ( DNA Pol-a) has been studied by
  NMR and CD.
• Recent cis-Platin Publications
• a) M. Basu and S. Basu in Liposome Methods and
  Pryocols (2002) Micelles and Liposomes in
  Metabolic Enzymes and Glycolipid
  Glycosyltransferase Assays(edited by S. Basu and
  M. Basu) , Humana Press, New York
  pages,107-130
• b) R. N. Bose, D. Li, W. W. Yang, and S. Basu.
  (1999) J. Biomolecular Struct. Dynamics, 16,
  1075-1085. NMR Structures of a Nonapeptide from
  DNA Binding Domain of Human Pol-a
• b) R. N. Bose, D. Li, M. Kennedy, and S. Basu.
  (1995) J. Chem. Soc. Chem. Commun., 1731- a.
  1732. Facile Formation of a cis-Platin-Nonapeptid
  e-Complex of Human DNA Pol-a.

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TARGETED DELIVERY OF DRUGS TO CANCER CELLS (By
LIPOSOME BULLETS)
Mab (CSLEX)
SA-Lex
Synthetic Liposomes Containing Anti-cancer
Drugs/ cis-Platin, Tamoxifan, Melphalan,
Betulinic Acid L/D-PPMP,
L/D-PDMP, GD3, Ceramides, GD1b, GT1b
Metastatic Cancer Cells (Colon, Breast,Neuronal,
Prostate, ALL)
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Structure of Apoptotic Drugs
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APOPTOSIS AND DNA METABOLISM BY ANTI- CANCER
DRUGS(cis-Platin/L(D)-PPMP/GD3)
APOPTOSIS
DNA-REPLICATION
APOPTOSIS
DNA-REPLICATION
cis-Platin ?
UNREGULATED
INHIBITED
cis-Platin ?
REGULATED
?X
Y?
CASPASES
cis-Platin ?
Pol-a
Cyto-C
Cyto-C
NUCLEARDNA
NUCLEARDNA
MITOCHONDRIA
MITOCHONDRIA
DIFFERENTIATED NOMAL CELLS (n1)
UNDIFFERENTIATED TUMOR OR CANCER CELLS (2n)
(COLO-205/SKBR-3/PA-3/ALL)
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APOPTOSIS AND DNA METABOLISM BY ANTI-CANCER DRUGS
(cis-PLATIN // L-/D-PPMP // GD3)
cis-PLATIN
DNA-REPLICATION
APOPTOSIS
DNA-REPLICATION
APOPTOSIS
Pol-?
?
?
REGULATED
Y ?
X?
INHIBITED
Cyto-c
CASPASES
NUCLEAR DNA
Cyto-c
NUCLEAR DNA
MITOCHONDRION
DNA-DEGRADATION
MITOCHONDRION
DIFFERENTIATED NORMAL CELLS (N1)
UNDIFFERENTIATED TUMOR OR CANCER CELLS (
2N ) (COLO-205 / SKBR-3 / PA-3 / ALL)