HFE gene mutation and iron overload in Egyptian pediatric acute lymphoblastic leukemia survivors: a single-center study

Figures & data

Figure 1. Agarose gel electrophoresis of (1) amplification product of HFE gene at 387 bp. Lane 1 shows 50 bp ladder; lanes (2, 3, 4, 5, 6, 7, 8, 9, 10, and 11) show the amplification product of HFE gene at 387 bp. (2) Digestion products of HFE gene by RsaI at 247 and 140 bp. Lane 1 shows 50 bp ladder; lanes (2, 3, 4, 5, 6, 7, 8, 9, and 10) show GG homozygote (normal alleles) at (247–140) bp, lane 11 shows 100 bp ladder. (3) Amplification product of HFE gene at 208 bp. Lane (1) shows 100 bp ladder; lanes (2, 3, 4, 5, 6, 7, 8, 9, and 10) show the amplification product of HFE gene at 208 bp; lane (11) shows 50 bp ladder. (4) Digestion products of HFE gene by BclI at 70, 138, and 208 bp. Lane 1 shows 50 bp ladder; lanes (2, 6, 8, and 10) show CC homozygote at (70–138) bp, lanes (3, 4, 5, and 7) show GG homozygote at (208) bp and lanes (9 and 11) show CG heterozygote at (70–138 and 208) bp.

Table 1. Demographic data of the studied groups.

Parameter	Patients	Controls	Test type	p Value
Age (years)			1.1*
Range	4–18	4–18		>0.05
Mean ± SD	11.01 ± 4.6	9.6 ± 3.3
Sex			0**
Male	21, 60%	19, 60%		>0.05
Female	14, 40%	16, 40%

*U test (Mann–Whitney test).

**X², Chi square.

Table 2. Anthropometric measures of the studied groups.

	Patients, N = 35	Controls, N = 15	t test	p Value
Height (cm)
Range	96–177	98–160	0.942*	0.351
Mean ± SD	137.4 ± 23.6	130.8 ± 20.8		>0.05
Weight (kg)
Range	15–88	15.4–59	2.7*	0.009
Mean ± SD	41.6 ± 17	28.3 ± 12.2		<0 . 05
BMI
Range	13.85–33	16.25–26.5	2.6	0.012
Mean ± SD	22.9 ± 6.8	17.6 ± 5.7		<0.05

Bold, significant; BMI, body mass index.

U test (Mann–Whitney test).

*Significant.

Table 3. Iron profile of the studied groups.

	Patients	Controls	U test	p Value
Serum ferritin (ng ml^−^1)
Range	19.4–3540	30.99–88	2.8	0.006
Mean ± SD	737.6 ± 99 . 2	51.6 ± 18.2		<0.05
Serum iron (μg dl^−1)
Range	64–167	58–121	1.4	0.152
Mean ± SD	100.2 ± 36 . 4	85.8 ± 17.4		>0.05
TIBC (μg dl^−1)
Range	180–334	156–329	0.541	0.591
Mean ± SD	273.3 ± 72.7	261.9 ± 55.9		>0.05
TSI (%)
Range	32–50	32–37	2.2	0.03
Mean ± SD	46.9 ± 21.1	34.7 ± 1.7		<0.05

Bold, significant; U-test (Mann–Whitney test), TIBC, total iron binding capacity; TSI, transferrin saturation index.

Table 4. Genotype frequencies and allelic distribution of C282Y and H63D mutations among studied groups.

	Patients		Controls		X^2	p Value
	N (35)	%	N(35)	%
C282Y genotypes
GG (wild homozygous)	35	100	15	100	–	–
H63D genotypes
CC (wild homozygous)	19	54.3	32	80	13.08	0.001*
CG (heterozygous mutant)	10	28.6	3	20
GG (homozygous mutant)	6	17.1	0	0
Allele frequency	N = 70		N = 30		Fisher exact	p Value
C	48	68.6	27	90		0 . 02
G	22	31.4	3	10	5 . 1	S

X², Chi-square test; S, significant; Bold, significant.

*Significant.

Table 5. Iron profile with different H63D genotypes in studied groups.

		H63D genotypes			f test	p Value	LSD
	Controls, N = 35	GG, N = 6	CG, N = 10	CC, N = 19
S. ferritin (ng ml^−1)
Range	30.99–88	430.9–3063	35.5–3540	19.4–803.2	6.1*	0.001	p^a  < 0 . 05, p^b < 0 . 05, p^c > 0.05, p^d  < 0 . 001, p^e > 0.05, p^f < 0 . 05
Mean ±SD	51.6 ± 18.2	1715.7 ± 1020.9	568.8 ± 99.3	323.9 ± 259		<0.05
S. iron (μg dl^−1)
Range	58–121	78–198	64–137	67–102	5.7	0.002	p^a < 0 . 05, p^b  < 0 . 05, p^c  > 0.05, p^d  < 0 . 05, p^e  > 0.05, p^f  > 0.05
Mean ± SD	85.8 ± 17.4	119.5 ± 52.1	93.3 ± 17.9	79.6 ± 11.6		<0.05
TIBC (μg dl^−1)
Range	156–329	160–331	182–307	180–334	2.1	0.103	p^a  < 0 . 05, p^b  > 0.05, p^c  > 0.05, p^d  > 0.05, p^e  > 0.05, p^f  > 0.05
Mean ± SD	261.9 ± 55.9	240.3 ± 71.9	232.1 ± 38.6	269.7 ± 46.1		>0.05
TSI (%)
Range	32–37	34–110	32–39	32–41	11.5	<0.001	p^a  < 0 . 001, p^b  < 0 . 001, p^c  > 0.05, p^d  < 0 . 001, p^e  > 0.05, p^f  > 0.05
Mean ± SD	34.7 ± 1.7	69.3 ± 30.4	34.8 ± 1.9	34.4 ± 2.1

*f test, one-way ANOVA; >0.05, non-significant; <0.05, significant; <0.001, highly significant.

Bold, significant; p^a, comparison between GG and CG; p^b, comparison between GG and CC; p^c, comparison between CG and CC; p^d, comparison between control and GG; p^e, comparison between control and CC; p^f, comparison between control and CG.

LSD, least standard deviation; CC, homozygote normal alleles; CG, heterozygote mutant alleles; GG, homozygote mutant alleles; TIBC, total iron binding capacity; TSI, transferrin saturation index.

Table 6. H63D genotype frequency among male and female ALL survivors.

H63D genotypes	Male		Female		X^2	p Value
	N(21)	%	N(14)	%
CC	13	61.9	6	42.9	2.3	0.31
CG	6	28.6	4	28.6
GG	2	9.5	4	28.6

Table 7. Iron profile among male and female ALL survivors.

	Male	Female	U test
S. ferritin (ng ml^−1)	471 ± 370.6	1137.5 ± 1092.3	2.2	0.03
S. iron (μg dl^−1)	91.2 ± 29.2	113.7 ± 42.9	1.84	0.07
TIBC (μg dl^−1)	283.4 ± 72.9	258.2 ± 72.3	1.004*	0.323
TSI (%)	41.7 ± 14.9	54.5 ± 26.9	1.81	0.08

*t test.

TIBC, total iron binding capacity; TSI, transferrin saturation index.