Percentage inhibition of Notch1, Notch2, and Hes1 expression by GSI
Patients, GSI (μM) . | Notch1 . | Notch2 . | Hes1 . | ||||
---|---|---|---|---|---|---|---|
P . | TM . | IC . | P . | TM . | IC . | ||
B-CLL 1 | |||||||
1 | 0 | 0 | 0 | 6 | 4 | 10 | 2 |
2.5 | 65 | 50 | 54 | 86 | 40 | 67 | 31 |
5 | 90 | 81 | 75 | 100 | 77 | 99 | 42 |
10 | 94 | 80 | 75 | 100 | 69 | 97 | 42 |
B-CLL 2 | |||||||
1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
2.5 | ND | ND | ND | ND | ND | ND | ND |
5 | 18 | 10 | 15 | 21 | 15 | 18 | 10 |
10 | 60 | 49 | 44 | 51 | 44 | 58 | 27 |
B-CLL 5 | |||||||
1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
2.5 | 51 | 40 | 46 | 60 | 35 | 44 | 29 |
5 | 70 | 57 | 61 | 75 | 51 | 73 | 46 |
10 | 71 | 66 | 70 | 96 | 89 | 96 | 49 |
B-CLL 6 | |||||||
1 | 0 | 13 | 14 | 0 | 0 | 0 | 0 |
2.5 | 54 | 49 | 47 | 42 | 48 | 51 | 51 |
5 | 70 | 71 | 64 | 87 | 58 | 96 | 82 |
10 | 98 | 86 | 81 | 99 | 57 | 98 | 87 |
B-CLL 8 | |||||||
1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
2.5 | 29 | 16 | 14 | 35 | 18 | 29 | 12 |
5 | 81 | 74 | 72 | 100 | 95 | 99 | 89 |
10 | 88 | 75 | 76 | 100 | 96 | 100 | 90 |
B-CLL 14 | |||||||
1 | ND | ND | ND | ND | ND | ND | ND |
2.5 | 61 | 50 | 51 | 77 | 46 | 54 | 41 |
5 | 92 | 75 | 78 | 95 | 80 | 93 | 66 |
10 | 94 | 74 | 80 | 91 | 87 | 100 | 66 |
B-CLL 17 | |||||||
1 | 0 | 0 | 0 | 1 | 2 | 4 | 0 |
2.5 | 39 | 25 | 29 | 44 | 31 | 35 | 29 |
5 | 93 | 79 | 82 | 100 | 81 | 87 | 59 |
10 | 100 | 83 | 85 | 100 | 100 | 100 | 63 |
B-CLL 20 | |||||||
1 | 5 | 7 | 0 | 4 | 8 | 6 | 0 |
2.5 | 62 | 61 | 43 | 67 | 50 | 57 | 32 |
5 | 85 | 75 | 72 | 100 | 65 | 100 | 60 |
10 | 90 | 80 | 81 | 100 | 82 | 100 | 64 |
B-CLL 21 | |||||||
1 | 13 | 15 | 16 | 17 | 10 | 21 | 15 |
2.5 | 21 | 18 | 24 | 18 | 11 | 20 | 15 |
5 | 78 | 60 | 63 | 100 | 56 | 95 | 49 |
10 | ND | ND | ND | ND | ND | ND | ND |
B-CLL 22 | |||||||
1 | 26 | 21 | 29 | 30 | 17 | 31 | 20 |
2.5 | 31 | 25 | 30 | 35 | 21 | 40 | 33 |
5 | 81 | 73 | 69 | 99 | 99 | 98 | 70 |
10 | 91 | 85 | 83 | 100 | 100 | 100 | 90 |
B-CLL 23 | |||||||
1 | 0 | 0 | 0 | 5 | 3 | 7 | 0 |
2.5 | 25 | 21 | 21 | 16 | 14 | 23 | 10 |
5 | 83 | 65 | 64 | 89 | 71 | 93 | 46 |
10 | 88 | 77 | 79 | 100 | 88 | 100 | 54 |
B-CLL 24 | |||||||
1 | ND | ND | ND | ND | ND | ND | ND |
2.5 | ND | ND | ND | ND | ND | ND | ND |
5 | 92 | 80 | 77 | 100 | 81 | 96 | 39 |
10 | 100 | 97 | 96 | 100 | 84 | 99 | 52 |
Patients, GSI (μM) . | Notch1 . | Notch2 . | Hes1 . | ||||
---|---|---|---|---|---|---|---|
P . | TM . | IC . | P . | TM . | IC . | ||
B-CLL 1 | |||||||
1 | 0 | 0 | 0 | 6 | 4 | 10 | 2 |
2.5 | 65 | 50 | 54 | 86 | 40 | 67 | 31 |
5 | 90 | 81 | 75 | 100 | 77 | 99 | 42 |
10 | 94 | 80 | 75 | 100 | 69 | 97 | 42 |
B-CLL 2 | |||||||
1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
2.5 | ND | ND | ND | ND | ND | ND | ND |
5 | 18 | 10 | 15 | 21 | 15 | 18 | 10 |
10 | 60 | 49 | 44 | 51 | 44 | 58 | 27 |
B-CLL 5 | |||||||
1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
2.5 | 51 | 40 | 46 | 60 | 35 | 44 | 29 |
5 | 70 | 57 | 61 | 75 | 51 | 73 | 46 |
10 | 71 | 66 | 70 | 96 | 89 | 96 | 49 |
B-CLL 6 | |||||||
1 | 0 | 13 | 14 | 0 | 0 | 0 | 0 |
2.5 | 54 | 49 | 47 | 42 | 48 | 51 | 51 |
5 | 70 | 71 | 64 | 87 | 58 | 96 | 82 |
10 | 98 | 86 | 81 | 99 | 57 | 98 | 87 |
B-CLL 8 | |||||||
1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
2.5 | 29 | 16 | 14 | 35 | 18 | 29 | 12 |
5 | 81 | 74 | 72 | 100 | 95 | 99 | 89 |
10 | 88 | 75 | 76 | 100 | 96 | 100 | 90 |
B-CLL 14 | |||||||
1 | ND | ND | ND | ND | ND | ND | ND |
2.5 | 61 | 50 | 51 | 77 | 46 | 54 | 41 |
5 | 92 | 75 | 78 | 95 | 80 | 93 | 66 |
10 | 94 | 74 | 80 | 91 | 87 | 100 | 66 |
B-CLL 17 | |||||||
1 | 0 | 0 | 0 | 1 | 2 | 4 | 0 |
2.5 | 39 | 25 | 29 | 44 | 31 | 35 | 29 |
5 | 93 | 79 | 82 | 100 | 81 | 87 | 59 |
10 | 100 | 83 | 85 | 100 | 100 | 100 | 63 |
B-CLL 20 | |||||||
1 | 5 | 7 | 0 | 4 | 8 | 6 | 0 |
2.5 | 62 | 61 | 43 | 67 | 50 | 57 | 32 |
5 | 85 | 75 | 72 | 100 | 65 | 100 | 60 |
10 | 90 | 80 | 81 | 100 | 82 | 100 | 64 |
B-CLL 21 | |||||||
1 | 13 | 15 | 16 | 17 | 10 | 21 | 15 |
2.5 | 21 | 18 | 24 | 18 | 11 | 20 | 15 |
5 | 78 | 60 | 63 | 100 | 56 | 95 | 49 |
10 | ND | ND | ND | ND | ND | ND | ND |
B-CLL 22 | |||||||
1 | 26 | 21 | 29 | 30 | 17 | 31 | 20 |
2.5 | 31 | 25 | 30 | 35 | 21 | 40 | 33 |
5 | 81 | 73 | 69 | 99 | 99 | 98 | 70 |
10 | 91 | 85 | 83 | 100 | 100 | 100 | 90 |
B-CLL 23 | |||||||
1 | 0 | 0 | 0 | 5 | 3 | 7 | 0 |
2.5 | 25 | 21 | 21 | 16 | 14 | 23 | 10 |
5 | 83 | 65 | 64 | 89 | 71 | 93 | 46 |
10 | 88 | 77 | 79 | 100 | 88 | 100 | 54 |
B-CLL 24 | |||||||
1 | ND | ND | ND | ND | ND | ND | ND |
2.5 | ND | ND | ND | ND | ND | ND | ND |
5 | 92 | 80 | 77 | 100 | 81 | 96 | 39 |
10 | 100 | 97 | 96 | 100 | 84 | 99 | 52 |
Notch1, Notch2, and Hes1 expression was analyzed by Western blot in B-CLL cells treated with GSI or DMSO for 24 hours. For each patient, the intensity of the bands corresponding to Notch1 and Notch2 (P, TM, and IC) and Hes1 was calculated by densitometric analysis and normalized to β-actin levels. Values represent the percentage inhibition of Notch1, Notch2, and Hes1 expression in GSI-treated compared with DMSO-treated B-CLL cells. P indicates precursor; ND, not determined.