phd student – liana ramishvili scientific supervisor - prof . n. kotrikadze
DESCRIPTION
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 - PowerPoint PPT PresentationTRANSCRIPT
Iv. Javakhishvili Tbilisi State UniversityFaculty of Exact and Natural Sciences
Department of BiologyDivision 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
NormalEpithelial Cells of Prostate
Partial Activity of Krebs Cycle
Low level of Respiration and
Terminal Oxidation
Energetically InefficientCan only produce small
amount of Electorns
Low Levels of Reactive oxygen Species (ROS)
Mitochondria
Mitochondria
Incresed production of Reactive Oxygen
Species (ROS) Intensification of
Freeradical Processes
Mitochondrial Defects
Krebs Cycle Functions Properly
Malignant transformation of Epithelial Cells of Prostate
Increased electons Flow to the Electon transport Chain
Increased rate of Mitochondrial DNA Mutations
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.
The Goal of the Work
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 FluorescenceSpectroscopic Methods
Histo-morphological pictures of Prostate Tumours. A. Controle group B Benign Hyperplasia C. Benign Hyperplasia with PIN regions D. Prostate adenocarcinoma
A B
C D
TeTr i si naT l i s wyar o
F L1
N2-l az er i
L2
Ar+ - l az er i
YAG: Nd3+ - l az er i
CCD speqt r omet er i
FM
2
3
4
Ch
PM
IBM PC: monacemTa
Segr oveba da damuSaveba
ni muSebi s sadgami
Laser Canceroscope
Consists of 4 blocks :
•Lights Source block;
•Sample block
•Registration block
•Data collecting and processing block.
Excitation was carried out by N2 laser:
=337nm wavelength.
Recording of Spectra was carried out in the 300- 500 nm wavelength region.
Light source
Sample
Data collection and
processing
N2 Laser
registration
The Study of Prostate Tumour tissue by Laser induced Fluorescence
Tumour tissue fluorescence spectrum of men with prostate benign hyperplasia
λ (nm)
440-460nm (I=0,48)
390-400nm (I=0,38)
Flu
ores
c en
ce in
ten
sity
(I
)
Tumour tissue fluorescence spectrum of men with prostate benign hyperplasia with PING (3-4) regions
λ (nm)
440-460nm (I=0,9)
400-410nm (I-=0,65)
Flu
ores
cen
ce in
ten
sit y
(I)
460-470nm (I=0,8)
400-410nm (I=0,45)
Tumour tissue fluorescence spectrum of men with prostate
adenocarcinoma
λ (nm)
Flu
ores
cen
ce in
ten
sit y
(I
)
460-470nm (I=0,8)
400-410nm (I=0,45)
λ (nm)
Flu
ores
cen
ce in
ten
sit y
(I)
(I)
λ (nm)
440-460nm (I=0,48)
390-400nm (I=0,38)
Flu
ores
c en
ce in
ten
sity
(I) Benign tumour
Benign tumour with
PING(3-4) regions
Prostate Cancer
0
0.2
0.4
0.6
0.8
1
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
0
5
10
15
20
25
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.
0
20
40
60
80
100
120
140
160
1 2 3
1- Prostate Benign Hyperplasia;
2- Prostate Benign Hyperplasia with PING3-4 regions;
3- Prostate Cancer.
The Activity of Cytochromeoxidase
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.
m-aconitase
oxidates
NADH-Dehydrogenase
(I complex)
GSH
Citrate
Isocitrate
Enhanced Electron transfer
O2-
H2O2O2
Reduced Electron transfer
2H´+ 1/2O2
H20
Isocitrate DehydrogenaseNADPHNADP+
Succinate Dehydrogenase(SDH)
(II complex)
Ubiquinone/Cytochromeb(III complex)
Cytochrome Oxidase(COX)
(IV complex)
GSSGH2O + O2
Krebs Cycle
activation
Enhanced production
activates
Glutathione reductase
Glutathione peroxidase
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).
Glutathione-dependent Enzymes : Glutathione peroxidase (GSH-Px);
Glutathione reductase (GR);
Reduced Glutathione (GSH).
The Activity of Glutathione Peroxidase
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
1 2 3
1 2 3
1- Prostate Benign Hyperplasia;
2- Prostate Benign Hyperplasia with PING3-4 regions;
3- Prostate Cancer.
The Activity of Glutathione Reductase
0
0,005
0,01
0,015
0,02
0,025
0,03
0,035
1 2 3
1- Prostate Benign Hyperplasia;
2- Prostate Benign Hyperplasia with PING3-4 regions;
3- Prostate Cancer.
The Amount of Reduced Glutathione
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1 2 3
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).
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.
Conclusions:
m-aconitase
oxidates
NADH-Dehydrogenase
(I complex)
GSH
Citrate
Isocitrate
Enhanced Electron transfer
O2-
H2O2O2
Reduced Electron transfer
2H´+ 1/2O2
H20
Isocitrate DehydrogenaseNADPHNADP+
Succinate Dehydrogenase(SDH)
(II complex)
Ubiquinone/Cytochromeb(III complex)
Cytochrome Oxidase(COX)
(IV complex)
GSSGH2O + O2
Krebs Cycle
activation
Enhanced production
activates
Glutathione reductase
Glutathione peroxidase
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).
Thank you for attention
winamdebare jirkvlis lokalizacia da zonaluri anatomia CZ – centraluri zona; PZ – periferiuli zona; TZ – gardamavali
zona
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
prostatis epiTeliumSi arCeven bazalur, sekretorul da parakrinul-endokrinul ujredebs
prostatis keTilTvisebiani hiperplazia prostatis adenokarcinoma