Expression level of DEK in chronic lymphocytic leukemia is regulated by fludarabine and Nutlin-3 depending on p53 status

Figures & data

Table 1. Clinical and biological characteristics of 65 patients with chronic lymphocytic leukemia

Characteristics	Value (%)
Gender
Male	43 (66.2)
Female	22 (33.8)
Age (years)
≥ 60	33 (50.8)
< 60	32 (49.2)
Binet stages
Binet A	30 (46.2)
Binet B or C	35 (53.8)
LDH (n = 45)
≥ 250U/L	12 (26.7)
< 250U/L	33 (73.3)
β2-MG (n = 41)
≥ 3 mg/L	25 (61.0)
< 3 mg/L	16 (39.0)
CD38 (n = 63)
> 30%	22 (34.9)
≤ 30%	41 (65.1)
ZAP-70 (n = 63)
> 20%	14 (22.2)
≤ 20%	49 (77.8)
Del(13q14) (n = 53)
Positive	19 (35.8)
Negative	34 (64.2)
Trisomy 12 (n = 50)
Positive	15 (30.0)
Negative	35 (70.0)
Del(17p13) (n = 54)
Positive	9 (16.7)
Negative	45 (83.3)
IgH rearrangements (n = 47)
Positive	11 (23.4)
Negative	36 (76.6)
Del(11q22.3) (n = 57)
Positive	9 (15.8)
Negative	48 (84.2)
IGHV mutational status (n = 59)
Mutated	44 (74.6)
Unmutated	15 (25.4)
p53 mutations (n = 58)
Mutated	11 (19.0)
Unmutated	47 (81.0)

Abbreviations: β2-MG, β 2-microglobulin; IGHV, heavy chain variable region; LDH, lactate dehydrogenase.

Table 2. The differences of DEK mRNA expression level between various groups of patients

Clinical features	DEK [M (P5−P95)]	p value
Age (years)		0.773
≥ 60	7.815 × 10^−2 (1.712 × 10^−2−3.828 × 10^−2)
< 60	8.390 × 10^−2 (2.238 × 10^−2−5.455 × 10^−1)
Binet stages		0.612
A	7.757 × 10^−2 (2.903 × 10^−2−2.364 × 10^−2)
B+C	8.407 × 10^−2 (1.841 × 10^−2−6.077 × 10^−1)
LDH (n = 45)		0.115
≥ 250U/L	9.121 × 10^−2 (1.438 × 10^−2−2.535 × 10^−1)
< 250U/L	6.255 × 10^−2 (2.20 × 10^−2−4.989 × 10^−1)
β2-MG (n = 41)		0.33
≥ 3 mg/L	9.016 × 10^−2 (1.771 × 10^−2−4.029 × 10^−1)
< 3 mg/L	6.987 × 10^−2 (2.281 × 10^−2−5.885 × 10^−1)
CD38 (n = 63)		0.047
≤ 30%	9.333 × 10^−2 (2.210 × 10^−2−6.029 × 10^−1)
> 30%	7.563 × 10^−2 (1.835 × 10^−2−3.713 × 10^−1)
ZAP-70 (n = 63)		0.552
≤ 20%	8.098 × 10^−2 (1.738 × 10^−2−4.531 × 10^−1)
> 20%	8.205 × 10^−2 (2.247 × 10^−2−5.407 × 10^−1)
Del(13q14) (n = 53)		0.159
Positive	6.562 × 10^−2 (1.745 × 10^−2−3.861 × 10^−1)
Negative	9.190 × 10^−2 (2.247 × 10^−2−6.364 × 10^−1)
Trisomy 12 (n = 50)		0.546
Positive	7.962 × 10^−2 (3.443 × 10^−2−5.407 × 10^−1)
Negative	8.520 × 10^−2 (2.128 × 10^−2−6.364 × 10^−1)
Del(17p13) (n = 54)		0.006
Positive	1.374 × 10^−1 (2.256 × 10^−2−3.201 × 10^−1)
Negative	7.405 × 10^−2 (1.919 × 10^−2−5.885 × 10^−1)
IgH rearrangements (n = 47)		0.883
Positive	7.778 × 10^−2 (3.126 × 10^−2−1.103 × 10^−1)
Negative	8.935 × 10^−2 (2.241 × 10^−2−6.449 × 10^−1)
Del(11q22.3) (n = 57)		0.498
Positive	1.070 × 10^−1 (2.256 × 10^−2−3.861 × 10^−1)
Negative	7.899 × 10^−2 (1.607 × 10^−2−7.126 × 10^−1)
IGHV mutation status (n = 59)		0.025
Mutated	1.115 × 10^−2 (1.724 × 10^−2−2.264 × 10^−1)
Unmutated	6.976 × 10^−2 (2.386 × 10^−2−6.021 × 10^−1)
p53 mutations (n = 58)		0.655
Mutated	7.513 × 10^−2 (1.438 × 10^−2−1.142 × 10^−1)
Unmutated	8.55 × 10^−2 (2.350 × 10^−2−4.406 × 10^−1)

Abbreviations: β2-MG, β 2-microglobulin; IGHV, heavy chain variable region; LDH, lactate dehydrogenase.

Table 3. Clinical and biological characteristics in 22 CLL patients with cell culture

No.	Geneder	Age (years)	Binet stages	IGHV mutation status	Fludarabine-treated		p53 mutation status	p53 deletion	ATM deletion
					p53	p21
1	Female	78	A	Mutated	Yes	Yes	Wild type	No	No
2	Female	57	B	Unmutated	No	No	Mutated	Yes	Yes
3	Female	62	C	Mutated	Yes	Yes	Wild type	No	No
4	Male	83	B	Mutated	Yes	No	Mutated	Yes	No
5	Female	62	C	Mutated	Yes	No	Wild type	No	No
6	Female	52	C	Unmutated	Yes	Yes	Wild type	No	Yes
7	Male	48	C	Unmutated	No	No	Mutated	Yes	No
8	Male	58	C	Unmutated	No	Yes	Wild type	No	No
9	Male	50	A	Mutated	Yes	Yes	Wild type	No	No
10	Female	54	B	Mutated	Yes	Yes	Wild type	No	No
11	Male	80	C	Unmutated	Yes	Yes	Wild type	No	No
12	Female	70	B	Unmutated	No	No	Wild type	No	No
13	Female	67	A	Mutated	Yes	Yes	Wild type	No	No
14	Female	71	A	Unmutated	Yes	Yes	Wild type	No	No
15	Male	52	A	Unmutated	Yes	Yes	Wild type	No	No
16	Male	55	A	Mutated	Yes	Yes	Wild type	No	Yes
17	Female	61	C	Mutated	No	No	Wild type	No	No
18	Male	68	A	Mutated	Yes	No	Mutated	No	No
19	Male	53	C	Mutated	Yes	Yes	Wild type	No	No
20	Male	49	C	Unmutated	Yes	No	Mutated	Yes	No
21	Female	64	A	Unmutated	Yes	Yes	Wild type	Yes	No
22	Female	57	A	Mutated	Yes	Yes	Wild type	No	Yes

“Yes” stands for raise up or positive; “No” stands for no change or negative.

Figure 1. The correlation between the p53 status and the level of DEK expression. In the primary CLL cells with normal p53 function (A and B), or without deletion (E and F) or mutation (I and J) of p53, DEK expression was significantly decreased after 24 h treatment with fludarabine or Nutin-3 compared with the cells in medium only. However, this downregulation of DEK expression was not observed in the primary CLL cells with p53 dysfunction (C and D), or deletion (G and H) or mutation (K and L) of p53.

Table 4. The sequences of qRT-PCR primers of DEK and β-actin

Primer		Sequence	length of product
DEK	forward	5‘-TCCAAAGCCTTCTGGCAAACCATT-3′	214 bp
	reverse	5‘-TGGTGGCTCCTCTTCACTTTCTTTA-3′
β-actin	forward	5′-AGCGAGCATCCCCCAAAGTT-3′	285 bp
	reverse	5′-GGGCACGAAGGCTCATCATT-3′