48r- shp16 + 2 gy
Post on 24-Feb-2016
50 Views
Preview:
DESCRIPTION
TRANSCRIPT
dNTP Supply Gene Expression Patterns in Microarrays after P53 Loss
Tom Radivoyevitch
Collaborative work with: John Pink, Charles Kunos, Gina Ferris, Ian Lent, Mark Jackson, Damian Junk
Pre-Rx 30’ 1 2 4 6 24
X X X X X XX X X X X
2X X X X X X XX X X X X
2X X X X X X X
48R-shp16 + 2 Gy
48R-shp16 + 2 Gy & 3-AP
CaSki + 2 Gy
CaSki + 2 Gy & 3-AP
TIME (hrs)
48R-shp16-shp53 + 2Gy
32 U219 CHIPS CaSki Cells
human mammary epithelial cells (HMEC)
9.4
10.2
RRM1 average of 4
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
10.0
11.5
RRM2 average of 2
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
7.0
8.5
RRM2B average of 3
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
P53 out so less p53R2
R1 compensates for p53R2 loss
No R2 compensation for p53R2 loss
Averaging probesets with aves > 6
911
CDKN1A average of 2
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
7.5
8.5
DCK average of 5
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
9.8
10.2
DCTD average of 5
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
P21 is p53 dependent, so there is a big loss in expression without it
Compensate for loss of de novo with increase in salvage
Lower average is consistent lower DCTD in next slide, i.e. less de novo dTMP
Lower levels is consistent with lower DCTD
Salvage compensates for less p53R2?Since R2 did not increase, not likely an S-fraction change
HPV does more than take down p53 9.
010
.5TK1 average of 2
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
10.5
11.5
TYMS average of 2
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
10.2
10.8
DGUOK average of 1
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
5.5
6.5
7.5
PPARG average of 2
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
6.0
7.0
TK2 average of 3
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
More Warburg => less mtDNA => less TK2
8.5
10.0
RRM1 average of 2
Epithelial StromalNormal Li-Fraumeni Li-Frau-Like Normal Li-Fraumeni Li-Frau-Like
610
RRM2 average of 2
Epithelial StromalNormal Li-Fraumeni Li-Frau-Like Normal Li-Fraumeni Li-Frau-Like
6.5
8.0
RRM2B average of 1
Epithelial StromalNormal Li-Fraumeni Li-Frau-Like Normal Li-Fraumeni Li-Frau-Like
Herbert et al. Oncotarget. 2010, 1(6): 405–422 (Normal, Li-Fraumeni, & LF-Like) 3 people, 4 replicates, 2 tissues (Breast epithelial and stromal cells)
Herbert et al. Oncotarget. 2010, 1(6): 405–422 (Normal, Li-Fraumeni, & LF-Like) 3 people, 4 replicates, 2 tissues (Breast epithelial and stromal cells)
911
CDKN1A average of 1
Epithelial StromalNormal Li-Fraumeni Li-Frau-Like Normal Li-Fraumeni Li-Frau-Like
7.5
9.0
DCK average of 1
Epithelial StromalNormal Li-Fraumeni Li-Frau-Like Normal Li-Fraumeni Li-Frau-Like
9.9
10.3
DCTD average of 3
Epithelial StromalNormal Li-Fraumeni Li-Frau-Like Normal Li-Fraumeni Li-Frau-Like
68
10TK1 average of 2
Epithelial StromalNormal Li-Fraumeni Li-Frau-Like Normal Li-Fraumeni Li-Frau-Like
79
11
TYMS average of 2
Epithelial StromalNormal Li-Fraumeni Li-Frau-Like Normal Li-Fraumeni Li-Frau-Like
9.0
10.0 DGUOK average of 2
Epithelial StromalNormal Li-Fraumeni Li-Frau-Like Normal Li-Fraumeni Li-Frau-Like
5.0
6.5
8.0 PPARG average of 1
Epithelial StromalNormal Li-Fraumeni Li-Frau-Like Normal Li-Fraumeni Li-Frau-Like
7.6
8.2
TK2 average of 4
Epithelial StromalNormal Li-Fraumeni Li-Frau-Like Normal Li-Fraumeni Li-Frau-Like
p53R2 Inhibits the Proliferation of Human Cancer Cells in Association with Cell-Cycle Arrest Keqiang Zhang, Jun Wu, Xiwei Wu, Xiaochen Wang, Yan Wang, Ning Zhou, Mei-ling Kuo, Xiyong Liu, Bingsen Zhou, Lufen Chang, David Ann, and Yun Yen Mol Cancer Ther; 10(2) 2011Affy Gene ST 1.0. GEO gse25238 (p53R2 siRNA of oropharyngeal carcinoma KB cells)
8.2
8.6
CDKN1A
Cntrls siP53R2
6.70
6.80
6.90
DCK
Cntrls siP53R2
7.8
8.0
8.2
8.4
DCTD
Cntrls siP53R2
10.2
510
.35
10.4
5 RRM1
Cntrls siP53R2
8.2
8.6
9.0
RRM2
Cntrls siP53R2
5.0
6.0
7.0
RRM2B
Cntrls siP53R2
9.90
10.0
0
TK1
Cntrls siP53R2
9.0
9.1
9.2
9.3
TYMS
Cntrls siP53R2
7.5
8.0
8.5
9.0
DCK Medians: 8.3 (p16); 9.12 (p53) Wilcoxon P = 9.83e-06
9.6
10.0
10.4
UCK2 Medians: 10.2 (p16); 10.5 (p53) Wilcoxon P = 0.0199
=> Rationale for using Deoxy-Aza-Cytidine (DAC) instead of aza-cytidine in p53 minus myelodysplastic syndrome (MDS) patients
Ongoing Active Research Projects1. Optimize triapine timing relative to irradiation of p53- cervical cancers (Charles Kunos).
2. Treatment of MDS with DAC (Yogen Saunthararajah, Cleveland Clinic). Goal is to understand dNTP supply system dependence on gender, age, and p53 status.
0 5 10 15 20
0.8
1.2
time
3 Gy10 Gy
RRM1
0 5 10 15 20
0.4
0.7
1.0
time
RRM2
0 5 10 15 20
1.0
1.4
1.8
time
TYMS
0 5 10 15 20
0.6
1.0
time
MTHFD1
0 5 10 15 200.
71.
01.
3
time
TK1
0 5 10 15 20
1.0
2.5
4.0
time
CDKN1A
GEO data GDS479Jen, K. Y. and V. G. Cheung (2003). Transcriptional response of lymphoblastoid cells to ionizing radiation. Genome Res 13(9): 2092-100.
Xiao-Peng Zhang, Feng Liu, and Wei Wang, (2011) PNAS 108, 8990–8995
Nor
mal
ized
to t=
0
Hours after irradiation
46
8
NME4 average of 1
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
57
NME5 average of 1
Ca.Pre
Ca.1 Ca.2 Ca.4 Ca.6Ca.2
4
CaAP.1
CaAP.2
CaAP.4
CaAP.6
CaAP.24
shp1
6
p16.P
re
p16.3
0Mp1
6.1p1
6.2p1
6.4p1
6.6
p16.2
4
p16A
P.1
p16A
P.2
p16A
P.4
p16A
P.6
p16A
P.24sh
p53
p53.P
re
p53.3
0Mp5
3.1p5
3.2p5
3.4p5
3.6
p53.2
4
NDPK expressed in NonMEtastatic cells, decreasing as cells become more cancerous
X48R Ca.1 Ca.2 Ca.24 Ca.4 Ca.6 Ca.Pre CaAP.1 CaAP.2 CaAP.24 CaAP.4 CaAP.6 Ep MIR
68
12
MYC OSM.Emt OSM.Ep p16.1 p16.2 p16.24 p16.30M p16.4 p16.6 p16.Pre p16AP.1 p16AP.2 p16AP.24
68
12
p16AP.4 p16AP.6 p53.1 p53.2 p53.24 p53.30M p53.4 p53.6 p53.Pre shp16 shp53 Stem1 Stem2
68
12
UDP
CDP
GDP
ADP
dTTP
dCTP
dGTP
dATP
dT
dC
dG
dADNA
dUMP
dU
TS
DCTD
dCK
DNA
poly
mer
ase
TK1
cytosol
mitochondria
dT
dC
dG
dA
TK2
dGK
dTMPdCMP
dGMP
dAMP
dTTPdCTP
dGTP
dATP
5NT
NT2
cytosol
nucleus
dUDP
dUTPdUTPase
dN
dN
dCK
flux activation inhibition
ATPordATP
RNR
dCK
dNTP Supply
top related