Background
Methods
Patient samples
Cytospin preparation
Cell cultures
Immunofluorescence assay
Evaluation of sensitivity and specificity of CTC detection
Evaluation of ALDH1 and TWIST expression in cancer cell lines using the ARIOL system
ALDH1 expression levels | HepG2 | SKBR3 | MCF7 | MDA.MB.231 | ||||
---|---|---|---|---|---|---|---|---|
Range
| Median ± SEa
|
Range
| Median ± SEa
|
Range
| Median ± SEa
|
Range
| Median ± SEa
| |
High
|
5 – 25
| 15 ± 0.25 |
10 – 25
| 15 ± 0.23 |
20 – 25
| 20 ± 0.11 |
15 – 25
| 20 ± 0.18 |
Low
|
30 – 55
| 45 ± 0.30 |
35 – 55
| 45 ± 0.29 |
35 – 55
| 45 ± 0.29 |
30 – 55
| 45 ± 0.29 |
Negative
|
60 – 90
| 70 ± 0.30 |
60 – 90
| 80 ± 0.39 |
60 – 90
| 75 ± 0.29 |
60 – 100
| 80 ± 0.46 |
Statistical analysis
Results
Sensitivity and specificity of CTC detection
Definition of high and low ALDH1 expression levels and characterization of TWIST sub-cellular localization in cancer cell lines
Expression of ALDH1 and TWIST in CTCs of patients with early breast cancer
ALDH1 expression
CTC phenotypes
| Patients (%) | Percentage of CTCs per patient (mean; range) | CTCs (%) | |||||
---|---|---|---|---|---|---|---|---|
Early
|
Metastatic
|
p value
|
Early
|
Metastatic
|
p value
|
Early
|
Metastatic
| |
ALDH1 high
| 30.8 | 80.0 |
0.009
| 23 (0–100) | 75 (0–100) |
0.001
| 38.7 | 83.5 |
ALDH1 low/neg
| 92.3 | 32.0 |
0.006
| 77 (0–100) | 25 (0–100) |
0.001
| 61.3 | 16.5 |
TWIST nuc
| 30.8 | 80.0 |
0.009
| 29 (0–100) | 73 (0–100) |
0.006
| 32.3 | 70.3 |
TWIST cyt/neg
| 76.9 | 40.0 |
0.040
| 71 (0–100) | 27 (0–100) |
0.006
| 67.7 | 29.7 |
TWIST expression
ALDH1 and TWIST co-expression
CTC phenotypes
| Patients (%) | Percentage of CTCs per patient (mean; range) | CTCs (%) | |||||
---|---|---|---|---|---|---|---|---|
Early
|
Metastatic
|
p value
|
Early
|
Metastatic
|
p value
|
Early
|
Metastatic
| |
ALDH1high / TWISTnuc
| 15.4 | 76.0 |
0.001
| 6 (0–50) | 64 (0–100) |
0.000
| 12.9 | 61.5 |
ALDH1high / TWISTcyt/neg
| 23.1 | 24.0 |
1.000
| 17 (0–100) | 11 (0–100) |
0.746
| 25.8 | 22.0 |
ALDH1low/neg / TWISTnuc
| 30.8 | 12.0 |
0.330
| 23 (0–100) | 8 (0–100) |
0.152
| 19.4 | 8.8 |
ALDH1low/neg / TWISTcyt/neg
| 61.5 | 20.0 |
0.078
| 54 (0–100) | 16 (0–100) |
0.026
| 41.9 | 7.7 |
Patients | ALDH1high/TWISTnuc | ALDH1high/TWISTcyt/neg | ALDH1low/neg/TWISTnuc | ALDH1low/neg/TWISTcyt/neg | |||||
---|---|---|---|---|---|---|---|---|---|
Early
|
Total CTC No
|
CTC No
(%)
|
CTC No
(%)
|
CTC No
(%)
|
CTC No
(%)
| ||||
1
|
3
|
0
| (0) |
0
| (0) |
0
| (0) |
3
| (100) |
2
|
5
|
0
| (0) |
0
| (0) |
0
| (0) |
5
| (100) |
3
|
3
|
1
| (33.3) |
0
| (0) |
2
| (66.7) |
0
| (0) |
4
|
6
|
3
| (50) |
1
| (16.7) |
2
| (33.3) |
0
| (0) |
5
|
6
|
0
| (0) |
6
| (100) |
0
| (0) |
0
| (0) |
Metastatic
| |||||||||
1
|
2
|
2
| (100) |
0
| (0) |
0
| (0) |
0
| (0) |
2
|
11
|
10
| (90.9) |
1
| (9.1) |
0
| (0) |
0
| (0) |
3
|
2
|
1
| (50) |
0
| (0) |
1
| (50) |
0
| (0) |
4
|
3
|
1
| (20) |
2
| (80) |
0
| (0) |
0
| (0) |
5
|
21
|
5
| (23.8) |
14
| (66.7) |
0
| (0) |
2
| (9.5) |
6
|
2
|
0
| (0) |
0
| (0) |
0
| (0) |
2
| (100) |
7
|
11
|
3
| (27.2) |
2
| (18.2) |
6
| (54.5) |
0
| (0) |