Background
Patients and methods
Patients
Methods
Determination of p53 functionality
Tissue preparation for multiparameter flow cytometry and sorting
SNP array hybridization for allelic state determination
FISH
p53 IHC staining
TP53 mutation analysis
mRNA expression arrays
Statistical analysis
Results
Patients’ description
Characteristics | Total N (%) |
---|---|
Age
| |
50-59 | 14 (17) |
60-69 | 27 (33) |
70-79 | 24 (30) |
80-89 | 16 (20) |
Gender
| |
Male | 34 (41) |
Female | 45 (54) |
Tumor Location
| |
Right | 52 (63) |
Left | 31 (37) |
Stage
| |
I and II | 54 (65) |
III | 29 (35) |
MMR status
| |
MSS | 55 (67) |
MSI-H | 27 (33) |
Chr.17p allelic state
| |
AB | 39 (47) |
LOH | 9 (11) |
CNLOH | 11 (13) |
ALOH | 7 (8) |
AI | 3 (4) |
Multiple clones | 14 (17) |
DNA index
| |
0.95 – 1.05 | 35 (46) |
1.06 – 1.40 | 10 (13) |
1.41 – 1.95 | 31 (41) |
TP53
| |
wt | 22 (55) |
mut | 18 (45) |
IHC p53
| |
0 % | 10 (13) |
>0% - ≤25% | 35 (46) |
>25% | 31 (41) |
Median Follow up in months (range)
| 68.84 (2–199) |
Allelic state
Predicted p53 functionality
p53 non functional N (%)
|
p53 functional N (%)
| p value | |
---|---|---|---|
TP53 mutational status
| |||
wt | 7 (33) | 14 (78) |
0.01
|
mut | 14 (67) | 4 (22) | |
P53 IHC
| |||
0 | 3 (11) | 7 (24) |
<0.001#
|
0 - ≤25% | 1 (3) | 22 (76) | |
>25% | 24 (86) | 0 (0) | |
Chr. 17 p status
| |||
AB | 5 (18) | 22 (76) |
<0.001*
|
LOH | 2 (7) | 4 (14) | |
Copy neutral LOH | 9 (32) | 0 (0) | |
Amplified LOH | 5 (18) | 1 (3) | |
Allelic Imbalance | 1(4) | 0 (0) | |
Two clones | 6 (21) | 2 (7) | |
Age category
| |||
50 – 59 | 4 (14) | 6 (22) | NS |
60 – 69 | 10 (36) | 9 (32) | |
70 – 79 | 10 (36) | 9 (32) | |
80 – 89 | 4 (14) | 4 (14) | |
DNA index
| |||
0.95 – 1.05 | 6 (22) | 21 (78) |
<0.001¶
|
1.06 – 1.4 | 4 (15) | 3 (11) | |
1.41 – 1.95 | 17 (63) | 3 (11) | |
MMR status
| |||
MSI | 4 (14) | 14 (50) |
0.009
|
MSS | 24 (86) | 14 (50) | |
Gender
| |||
Male | 12 (43) | 18 (62) | NS |
Female | 16 (57) | 11 (38) | |
Tumor Location
| |||
Right | 10 (36) | 19 (66) |
0.035
|
Left | 18 (64) | 10 (34) | |
Stage
| |||
I and II | 14 (50) | 22 (76) | 0.06 |
III | 14 (50) | 7 (24) | |
Median Follow up in months
| 66.75 | 89.77 |
0.4
|
Gene name
|
Chr. position
|
Gene description
| p-value , p53 functional vs., p53 non functional |
---|---|---|---|
PRKCZ
| 1p36.33-p36.2 | Serine threonine kinase involved in several processes such as proliferation, differentiation and secretion. | 4.95E-04 |
↑non functional | |||
LMO3
| 12p12.3 | Lim domain only 3 (rhombotin like 2). Expression of LMO-3 represses p53 mediated mRNA expression of target genes. | 1.2E-02 |
↑non functional | |||
CDKN1A
| 6p21.2 | Cyclin dependent kinase inhibitor. Causes cell cycle arrest in the presence of DNA damage. | 1.3E-02↑functional |
PDE6A
| 5q31.2-q34 | Phosphodiesterase 6A, cGMP-specific, rod, alpha | 7.47E-02 |
↑non functional | |||
SIAH1
| 16q12 | Seven in absentia homolog 1. Involved in ubiquitination and proteosome related degradation of specific proteins like beta catenin. | 2.60E-02 |
↑non functional | |||
TPD52L2
| 20q13.2-q13.3 | Tumor protein D52 like 2. Expressed in childhood leukemia and testes. | 4.65E-02 |
↑non functional | |||
MDM2
| 12q14.3-q15 | MDM2 p53 binding protein homolog (Mouse) | 1.25E-02 |
↑ functional | |||
HMGCS1
| 5p14-p13 | 3-hydroxy 3-methylglutaryl-CoA synthase I | 1E-01 |
↑functional |
Survival analysis
Variables | HR | 95% CI | p value |
---|---|---|---|
p53 & CSNK1A1
| |||
p53 - & CSNK1A1 + and p53+ & CSNKA1A +/− | Referent | ||
p53 - CSNK1A1 -
|
4.74
|
1.47 – 15.34
|
0.009*
|
Tumor stage
| |||
I & II | Referent | ||
III
|
3.48
|
1.08 – 11.2
|
0.037*
|
Tumor location
| |||
Right | Referent | ||
Left | 0.92 | 0.32 – 2.67 | 0.58 |
Gender
| |||
Male | 0.92 | 0.32 – 2.97 | 0.88 |
Female | Referent | ||
MMR state
| |||
MSS | 0.43 | 0.097 – 1.91 | 0.27 |
MSI | Referent |