Iv. Javakhishvili Tbilisi State University
Faculty of Exact and Natural Sciences
Department of Biology
Division of Cellular and Molecular Biology
Prostate Cancer Epithelial Cells and The Changes That Take
Place During Their Malignant Transformation
PhD Student – Liana Ramishvili
Scientific supervisor - Prof. N. Kotrikadze
Normal
Epithelial Cells of Prostate
Partial Activity of
Krebs Cycle
Mitochondria
Low level of
Respiration and
Terminal Oxidation
Low Levels of Reactive
oxygen Species (ROS)
Energetically Inefficient
Can only produce small
amount of Electorns
Malignant transformation
of Epithelial Cells of Prostate
Krebs Cycle
Functions Properly
Mitochondria
Incresed production of
Reactive Oxygen
Species (ROS)
Increased electons Flow to
the Electon transport Chain
Intensification of
Freeradical Processes
Increased rate of Mitochondrial
DNA Mutations
Mitochondrial
Defects
The Goal of the Work
To study the metabolic changes that take place in prostate
epithelial cells during their malignant transformation.
Tasks:
• To study of prostate tumor tissue by fluorescence spectroscopy.
• To study the mitochondrial defects (respiratory chain enzymes and
gluthatione dependent system) in epithelial cells of prostate tumor.
Object of investigation:
Tumour tissue of patients with prostate tumours:
- Prostate benign hyperplasia;
- Prostate benign hyperplasia with PING(3-4) regions;
- Prostate Cancer.
Method of Investigation:
Laser induced Fluorescence
Spectroscopic Methods
A
B
D
C
Histo-morphological pictures of Prostate Tumours.
A. Controle group
B Benign Hyperplasia
C. Benign Hyperplasia with PIN regions
D. Prostate adenocarcinoma
wyar o
Light source
T eT r i si naT l i s
L1
L2
IBM
DataPC:
mo nac emT a
collection
Segr o vebaand
da
processing
d amuSaveba
F
-l az er i
NN2 Laser
2
Laser Canceroscope
Consists of 4 blocks :
•Lights Source block;
Ar+ - l az er i
•Sample block
Ch
•Registration block

2
YAG: Nd3+ l az er i
3
4
FM
CCD
speqt r o met er i
registration
ni muSebi s
sad gami
Sample
•Data collecting and processing block.
PM
Excitation was carried out by N2 laser:
=337nm wavelength.
Recording of Spectra was carried out in the 300500 nm wavelength region.
The Study of Prostate Tumour tissue by Laser induced
Fluorescence
440-460nm (I=0,48)
390-400nm (I=0,38)
λ (nm)
Tumour tissue fluorescence spectrum of men with prostate benign hyperplasia
440-460nm (I=0,9)
400-410nm
(I-=0,65)
λ (nm)
Tumour tissue fluorescence spectrum of men with prostate benign hyperplasia with
PING (3-4) regions
460-470nm (I=0,8)
400-410nm
(I=0,45)
λ (nm)
Tumour tissue fluorescence spectrum of men with prostate
adenocarcinoma
440-460nm (I=0,9)
400-410nm
(I-=0,65)
Benign tumour
Benign tumour with
PING(3-4) regions
440-460nm
(I=0,48)
390-400nm (I=0,38)
λ (nm)
460-470nm (I=0,8)
λ (nm)
400-410nm
(I=0,45)
Prostate Cancer
1
0.8
0.6
0.4
0.2
0
1
2
3
The Changes of NADH Fluorescennce peaks intensities tumor tissue of prostate (440-460 nm)
1- Prostate Benign Hyperplasia;
2- Prostate Benign Hyperplasia with PING3-4 regions;
3- Prostate Cancer.
Conclusion
• Sharply Increased intensity of the Nicotinamide Coenzymes peak
(440-460 nm) in benign prostate tumor with PING3-4 regions and in
prostate adenocarcinoma compared with benign tumor tissue spectra,
reflects the type of metabolism that is typical to prostate malignant tumor
cells.
The Study of Mitochondrial respiratory chain
enzymes (complex II and Complex IV )
The Activity of Succinatedehydrogenase
25
20
15
10
5
0
1
2
3
1- Prostate Benign Hyperplasia;
2- Prostate Benign Hyperplasia with PING3-4 regions;
3- Prostate Cancer.
The sharp increase in SD activity presumably
indicates on the enhanced electrons flow in
respiratory chain of mitochondria.
The Activity of Cytochromeoxidase
160
140
120
100
80
60
40
20
0
1
2
3
1- Prostate Benign Hyperplasia;
2- Prostate Benign Hyperplasia with PING3-4 regions;
3- Prostate Cancer.
The insignificant changes in COX activity
presumably indicates on the low level of
terminal oxidation.
Thus,
- Sharp increase of the activity of SDH (complex II);
- Insignificant changes of COX (complex IV) activity ;
These changes Presumably indicates to activation of Krebs cycle in
mitochondria and increase of electrons flow in respiration chain on the
one hand, and to impairment of the terminal oxidation of oxygen, on the
other.
Epithelial Cells of Prostate
malignant Tissue (PIN G3-4,, Cap)
General scheme of energy metabolism Possible
alterations in mitochondria of epithelial cells of
prostate malignant tissue
(BHP with PIN G3-4,regions, CaP).
Krebs Cycle
activation
m-aconitase
oxidates
Citrate
Isocitrate
GSH
Enhanced production
Isocitrate Dehydrogenase
NADP+
H2O + O2
O2 -
activates
NADHDehydrogenase
(I complex)
O2
Succinate Dehydrogenase
(SDH)
Glutathione
peroxidase
Glutathione
reductase
NADPH
GSSG
H2O2
Ubiquinone/Cytochromeb
(III complex)
Cytochrome Oxidase
(COX)
(II complex)
(IV complex)
Enhanced Electron
transfer
Reduced Electron
transfer
2H´+ 1/2O2
H20
Glutathione-dependent Enzymes :
 Glutathione peroxidase (GSH-Px);
 Glutathione reductase (GR);
 Reduced Glutathione (GSH).
The Activity of Glutathione Peroxidase
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
11
2
2
1- Prostate Benign Hyperplasia;
2- Prostate Benign Hyperplasia with PING3-4 regions;
3- Prostate Cancer.
33
The Activity of Glutathione Reductase
0,035
0,03
0,025
0,02
0,015
0,01
0,005
0
1
2
1- Prostate Benign Hyperplasia;
2- Prostate Benign Hyperplasia with PING3-4 regions;
3- Prostate Cancer.
3
The Amount of Reduced Glutathione
1- Prostate Benign Hyperplasia;
2- Prostate Benign Hyperplasia with PING3-4 regions;
3- Prostate Cancer.
Thus, sharp activation of mitochondrial antioxidant system,
(GSH-Px, GR) revealed in BHP with PING(3-4) regions and
malignant tumor epithelial cells, indicates to intensification of
defensive abilities of tumor cells.
(to withstand switching of the mitochondrial way of apoptosis,
induced by free radicals).
Conclusions:
 Thus, stimulation of the activity of SDH and retention of COX activity in epithelial cells of prostate
malignant tissue may be responsible for sharp activation of isocitrate dehydrogenase and
correspondingly, for significant accumulation of NADP(H).
 Laser Induced Fluorescence spectra have shown the incresead intensity of NADP(H) peak in case of
malignant tumor tissue that corresponds with investigations in Mitochondria of tumor epithelial cells.
 Accumulation of NADP(H). may stipulate a sharp activation of the glutathione-depended system, which
was proved by our investigations. Activation of the GSH-dependent system (GSH-Px, GR) presumably
would be responsible for resistance of cancer cells against the oxidative stress.
 Changes in the activity of enzymes of the II and IV complexes of mitochondrial respiration chain and
antioxidant system, in case of prostate malignant trabsformation, are reflection of specific metabolic
changes in mitochondria.
 All the Above mentioned indicates to resistance of prostate malignant cells and correspondingly, to
intensification of proliferation processes.
Epithelial Cells of Prostate
malignant Tissue (PIN G3-4,, Cap)
General scheme of energy metabolism Possible
alterations in mitochondria of epithelial cells of
prostate malignant tissue
(BHP with PIN G3-4,regions, CaP).
Krebs Cycle
activation
m-aconitase
oxidates
Citrate
Isocitrate
GSH
Enhanced production
Isocitrate Dehydrogenase
NADP+
H2O + O2
O2 -
activates
NADHDehydrogenase
(I complex)
O2
Succinate Dehydrogenase
(SDH)
Glutathione
peroxidase
Glutathione
reductase
NADPH
GSSG
H2O2
Ubiquinone/Cytochromeb
(III complex)
Cytochrome Oxidase
(COX)
(II complex)
(IV complex)
Enhanced Electron
transfer
Reduced Electron
transfer
2H´+ 1/2O2
H20
Thank you for attention
prostatis keTilTvisebiani simsivne
viTardeba prostatis kranialur
nawilSi ZiriTadad
periureTraluri jirkvlebidan
PIN ubnebiani keTilTvisebiani da avTvisebiani simsivneebi
(ukana kaudalur nawilSi)
mTavari samozne jirkvlovani epiTeliumis sekretoruli ujredebia
winamdebare jirkvlis lokalizacia da zonaluri anatomia
CZ – centraluri zona; PZ – periferiuli zona; TZ – gardamavali zona
prostatis epiTeliumSi arCeven bazalur, sekretorul da parakrinul-endokrinul ujredebs
prostatis keTilTvisebiani hiperplazia
prostatis adenokarcinoma