Relationships between cytokine (IL-6 and TGF-β₁) gene polymorphisms and chromosomal damage in hospital workers

Figures & data

Table 1. Demographic characteristics of studied groups.

Characteristics	Pathologists	Controls
Subjects	50	50
Sex
Males	23	23
Females	27	27
Age
Total sample
Mean years [±SD]	41.12 ± 8.08	39.86 ± 8.06
Range (years)	25–60	24–66
Males
Mean years [±SD]	44.87 ± 8.35	41.09 ± 1.86
Range (years)	28–60	27–66
Females
Mean years [±SD]	39.56 ± 7.36	38.18 ± 1.37
Range (years)	25–52	24–53
Years of employment
Total sample
Mean years [±SD]	10.36 ±7.40	11.34 ± 6.34
Range (years)	1–27	1–31
Males
Mean years [±SD]	13.39 ± 7.61	12.74 ± 7.82
Range (years)	2–24	2–31
Females
Mean years [±SD]	7.78 ± 6.26	10.15 ± 4.56
Range (years)	1–27	1–20

Table 2. Primers and annealing temperatures for cytokines gene polymorphisms analyzed in the present study.

dbSNP ID	Cytokine	Sequence	T ( °C)	PCR product size (bp)	Reference
rs909253	TNFα (-308, G > A) Antisense primer G-sense primer A-sense primer	5′-TCTCGGTTTCTTCTCCATCG-3′ 5′-ATAGGTTTTGAGGGGCATGG-3′ 5-AATAGGTTTTGAGGGGCATGA-3′	60	184	Perrey et al. (1999)
rs1800896	IL 10 -1082 (G > A) Antisense primer G-sense primer A-sense primer	5′-AGTGCCAACTGAGAATTTGG-3′ 5′-CTACTAAGGCTTCTTTGGGAG-3′ 5′-ACTACTAAGGCTTCTTTGGGAA-3′	60	258	Perrey et al. (1999)
rs1800871	IL-10 (-819, C > T) Antisense primer C-sense primer T-sense primer	5′-AGGATGTGTTCCAGGCTCCT-3′ 5′-CCCTTGTACAGGTGATGTAAC-3′ 5′-ACCCFFGTACAGGTGATGTAAT-3′	60	233	Perrey et al. (1999)
rs1800470	TGFβ1 (Codon 25, G > C) Antisense primer G-sense primer C-sense primer	5′-GGCTCCCGGTTCTGCACTC-3′ 5′-GTCTGACGCCTGGCCG-3′ 5′-GTGCTGACGCCTGGCCC-3′	60	233	Perrey et al. (1999)
rs1800471	TGFβ1 (Codon 10, C > T) Antisense primer C-sense primer T-sense primer	5′-TCCGTGGGATACTGAGACAC-3′ 5′-GCAGCGGTAGCAGCAGCG-3′ 5′-AGCAGCGGTAGCAGCAGCA-3′	60	241	Perrey et al. (1999)
rs1800795	IL-6 (-174, G > C) Antisense primer G-sense primer C-sense primer	5′-TCGTGCATGACTTCAGCTTTA-3′ 5′-AATGTGACGTCCTTTAGCATG-3′ 5′-AATGTGACGTCCTTTAGCATC-3′	60	190	Zakharyan et al. (2012)
	Internal Control 1 Internal Control 2	5′-GCCTTCCCAACCATTCCCTTA-3′ 5′-TCACGGATTTCTGTTGTGTTTC-3′	60	429	Perrey et al. (1999)

Table 3. Frequency of SCE and RI values in metaphases of lymphocytes from studied subjects.

Groups	N	NSM	SCE	SCE/NSM [± SE]	M1	M2	M3	RI [± SE]
Pathologists	50	2500	12991	5.196 ± 0.195^a	2245	1857	807	1.703 ± 0.051^b
Non-HHI	40	2000	9225	4.613 ± 0.114^c	1759	1515	632	1.671 ± 0.069
HFI	10	500	3766	7.532 ± 0.226^c	486	342	175	1.695 ± 0.087
Males	23	1150	6027	5.241 ± 0.250	975	888	393	1.709 ± 0.086
Females	27	1350	6984	5.159 ± 0.296	1270	969	414	1.648 ± 0.077
Controls	50	2500	10531	4.212 ± 0.179^a	1643	1972	1373	1.941 ± 0.027^b
Non-HHI	47	2350	9507	4.046 ± 0.162	1520	1852	1328	1.959 ± 0.026
HFI	3	150	1024	6.827 ± 0.221	123	120	45	1.660 ± 0.090
Males	23	1150	5139	4.469 ± 0.238	804	901	596	1.910 ± 0.030
Females	27	1350	5392	3.994 ± 0.260	839	1071	777	1.967 ± 0.042

^a,bp < 0.001.

^cp = 0.005.

N = Number of analyzed subjects; NSM = Number of scored metaphases; SCE = Sister chromatid exchanges; Metaphases; RI (Replication Index) = (M₁ + 2M₂ + 3M₃)/N, where M₁, M₂ and M₃ represent number of cells undergoing first, second, and third mitosis and N is total number of meta-phase scored.

Table 4. Frequencies of chromosomal aberrations (CA) and cells with aberrations (CAB) in metaphases of lymphocytes from subjects.

	CAs										CAs/NSM %	CAB/NSM %
Groups	N	NSM	B’	B”	Dic	AF	R	TR	Total CA	Total CAB	Mean [± SE]	Mean [± SE]
Pathologists	50	10000	110	25	5	18	0	1	159	150	0.0159 ± 0.002^a	0.0150 ± 0.002^b
Males	23	4600	61	11	0	11	0	0	83	76	0.0180 ± 0.003	0.0165 ± 0.002
Females	27	5400	49	14	5	7	0	1	76	74	0.0141 ± 0.002	0.0137 ± 0.002
Controls	50	10000	45	9	4	14	1	4	77	76	0.008 ± 0.001^a	0.008 ± 0.001^b
Males	23	4600	11	5	1	5	1	2	25	24	0.005 ± 0.001	0.005 ± 0.001
Females	27	5400	34	4	3	9	0	2	52	52	0.010 ± 0.001	0.010 ± 0.001

^a,bp < 0.001.

N = Number of analyzed subjects; NSM = Number of Scored Metaphases; B’: chromatid breaks; B”: chromosome breaks; Dic = Dicentric chromosome; AF = Acentric Fragments; R = Rings; TR = Tri- or Tetra-Radials; CAs = Chromosome Aberrations; CAB = cells with aberrations.

Table 5. Allele and genotype frequencies of cytokine gene polymorphisms among pathologists and control subjects.

	Pathologists				Controls
Genotypes	Observed Genotypes (Frequency)	Expected Genotypes (Frequency)	χ^2^aHWE df = 1	Alleles (frequency)	Observed Genotypes (Frequency)	Expected Genotypes (Frequency)	χ^2aHWE df = 1	Alleles (frequency)
TNF-α −308
G/G	29 (0.580)	31.21 (0.624)	0.156	G = 79 (0.790)	33 (0.660)	34.45 (0.689)	0.061	G = 83 (0.830)
G/A	21 (0.420)	16.56 (0.332)	1.172	A = 21 (0.2170)	17 (0.340)	14.11 (0.282)	0.592	A = 17 (0.170)
A/A	0 (0.000)	2.21 (0.044)	2.205		0 (0.000)	1.45 (0.029)	1.445
Total			3.533				2.098
IL 10 −1082
G/G	35 (0.700)	35.38 (0.706)	0.002	G = 84 (0.840)	29 (0.580)	29.65 (0.593)	0.014	G = 77 (0.770)
G/A	14 (0.280)	13.44 (0.269)	0.023	A = 16 (0.160)	19 (0.380)	17.71 (0.354)	0.094	A = 23 (0.230)
AA	1 (0.020)	1.28 (0.026)	0.061		2 (0.040)	2.65 (0.053)	0.157
Total			0.087				0.265
IL-10 (−8191)
C/C	37 (0.740)	36.98 (0.740)	0.000	G = 86 (0.860)	40 (0.800)	40.50 (0.810)	0.006	C = 90 (0.900)
C/T	12 (0.240)	12.04 (0.241)	0.000	A = 14 (0.140)	10 (0.200)	9 (0.180)	0.111	T = 10 (0.100)
T/T	1 (0.020)	0.98 (0.020)	0.000		0 (0.000)	0.50 (0.010)	0.500
Total			0.000				0.734
TGF-β1 Codon 25
G/G	41 (0.820)	41.41 (0.828)	0.004	G = 91 (0.910)	38 (0.760)	38.72 (0.775)	0.013	G = 88 (0.880)
G/C	9 (0.180)	8.19 (0.164)	0.080	C = 9 (0.090)	12 (0.240)	10.56 (0.211)	0.196	C= 12 (0.120)
C/C	0 (0.000)	0.41 (0.008)	0.405		0 (0.000)	0.72 (0.014)	0.720
Total			0.489				0.930
TGF-β1 Codon 10
C/C	9 (0.180)	11.05 (0.221)	0.379	C = 47 (0.470)	13 (0.260)	13.01 (0.260)	0.000	C = 51 (0.510)
C/T	29 (0.580)	24.91 (0.498)	0.672	T = 53 (0.530)	25 (0.500)	24.99 (0.500)	0.000	T = 49 (0.490)
T/T	12 (0.240)	14.05 (0.281)	0.298		12 (0.240)	12.01 (0.240)	0.000
Total			1.348				0.000
IL-6 (−174 G > C)
G/G	34 (0.680)	35.28 (0.706)	0.046	C = 84 (0.840)	32 (0.640)	33.62 (0.672)	0.078	C = 82 (0.820)
G/C	16 (0.320)	13.44 (0.269)	0.488	T = 16 (0.160)	18 (0.360)	14.76 (0.295)	0.711	T = 18 (0.180)
C/C	0 (0.000)	1.28 (0.026)	1.280		0 (0.000)	1.62 (0.032)	1.620
Total			1.814				2.409

^aChi-square values calculated from number of observed/expected genotypes; HWE = Hardy–Weinberg Equilibrium; df = degree freedom.

Table 6. Frequency of SCE and RI values in metaphases of lymphocytes from subjects – according to cytokine polymorphism.

Pathologists	N	NSM	SCEs	SCEs/NSM [± SE]	M1	M2	M3	RI [± SE]
IL10 (−1082)
(GG)	35	1750	9357	5.347 ± 0.244	1561	1290	536	1.643 ± 0.069
(GA)	14	700	3371	4.816 ± 0.328	613	540	269	1.786 ± 0.106
(AA)	1	50	263	5.260	71	27	2	1.310
IL10 (−819)
(CC)	37	1850	10114	5.467 ± 0.242	1583	1414	646	1.752 ± 0.057
(CT)	12	600	2637	4.395 ± 0.177	618	417	131	1.538 ± 0.109
(TT)	1	50	240	4.80	44	26	30	1.860
IL6
(GG)	34	1700	8806	5.180 ± 0.250^a	1538	1203	546	1.681 ± 0.064
(GC)	16	800	4185	5.231 ± 0.307^a	707	654	261	1.749 ± 0.083
TNF
(GG)	29	1450	7623	5.257 ± 0.284	1266	1063	567	1.756 ± 0.065
(GA)	21	1050	5368	5.112 ± 0.225	979	794	240	1.565 ± 0.088
TGFβ1 codon 10
(TT)	12	600	3104	5.173 ± 0.493^b	447	366	318	1.889 ± 0.132
(TC)	29	1450	7508	5.178 ± 0.256^c	1372	1137	120	1.639 ± 0.057
(CC)	9	450	2379	5.287 ± 0.328^b,c	426	354	120	1.660 ± 0.105
TGFβ1 codon 25
(GG)	41	2050	10655	5.198 ± 0.227	1902	1525	592	1.641 ± 0.060
(GC)	9	450	2336	5.191 ± 0.305	343	332	215	1.836 ± 0.152
IL10 (−1082)
(GG)	29	1450	6271	4.325 ± 0.254	912	1173	801	1.952 ± 0.034
(GA)	19	950	3792	3.992 ± 0.251	658	725	518	1.927 ± 0.042
(AA)	2	100	493	4.930 ± 1.470	77	76	48	1.865 ± 0.295
IL10 (−819)
(CC)	40	2000	8482	4.241 ± 0.203	1326	1589	072	1.930 ± 0.029
(CT)	10	500	2074	4.148 ± 0.419	321	385	295	1.976 ± 0.067
IL6
(GG)	32	1600	6623	4.139 ± 0.237	1037	1301	848	1.932 ± 0.028
(GC)	18	900	4056	4.507 ± 0.323	595	679	533	1.972 ± 0.051
TNF
(GG)	33	1650	6647	4.028 ± 0.201	1097	1267	937	1.952 ± 0.033
(GA)	17	850	3909	4.599 ± 0.334	550	707	430	1.914 ± 0.042
TGFβ1 codon 10
(CC)	13	650	2838	4.366 ± 0.358	496	489	316	1.863 ± 0.064
(CT)	25	1250	5337	4.270 ± 0.253	784	999	717	1.973 ± 0.030
(TT)	12	600	2381	3.968 ± 0.396	367	486	334	1.951 ± 0.058
IL10 (−8191)
(CC)	40	2000	8482	4.241 ± 0.203	1326	1589	072	1.930 ± 0.029
(CT)	10	500	2074	4.148 ± 0.419	321	385	295	1.976 ± 0.067

^aP = 0.017.

^b,cP = 0.021.

Table 7. Frequencies of chromosomal aberrations (CAs) and cells with aberrations (CAB) in lymphocytes of subjects – according to cytokine polymorphism.

Pathologists	N	NSM	B’	B’’	Dic	AF	R	TR	Total CAs	Total CAB	CAs/NSM Mean [± SE]	CAB/NSM Mean [± SE]
IL10 (−1082)
(GG)	35	7000	78	20	3	13	0	1	115	109	0.0164 ± 0.002	0.0156 ± 0.002
(GA)	14	2800	30	5	2	5	0	0	42	39	0.0150 ± 0.002	0.0147 ± 0.002
(AA)	1	200	2	0	0	0	0	0	2	2	0.010	0.010
IL10 (−819)
(CC)	37	7400	83	19	2	15	0	1	120	111	0.0162 ± 0.002	0.0150 ± 0.002
(CT)	12	2400	19	5	3	2	0	0	29	29	0.0121 ± 0.002	0.0121 ± 0.002
(TT)	1	200	8	1	0	1	0	0	10	10	0.050	0.050
IL6
(GG)	34	6800	70	19	3	16	0	1	109	102	0.0160 ± 0.002	0.0150 ± 0.002
(GC)	16	3200	40	6	2	2	0	0	50	48	0.0156 ± 0.003	0.0150 ± 0.003
TNF
(GG)	29	5800	65	16	2	10	0	0	93	88	0.0160 ± 0.002	0.0152 ± 0.002
(GA)	21	4200	45	9	3	8	0	1	66	62	0.0157 ± 0.002	0.0148 ± 0.002
TGFβ1 codon 10
(TT)	12	2400	31	4	1	6	0	0	42	37	0.0175 ± 0.004	0.0154 ± 0.003
(TC)	29	5800	60	15	4	10	0	1	90	86	0.0155 ± 0.002	0.0148 ± 0.002
(CC)	9	1800	19	6	0	2	0	0	27	27	0.0150 ± 0.003	0.0139 ± 0.002
TGFβ1 codon 25
(GG)	41	8200	86	15	5	14	0	0	120	114	0.0146 ± 0.002	0.0139 ± 0.001
(GC)	9	1800	24	10	0	4	0	1	39	36	0.0217 ± 0.006	0.0200 ± 0.005
IL10 (−1082)
(GG)	35	7000	78	20	3	13	0	1	115	1089	0.0164 ± 0.002	0.0156 ± 0.002
(GA)	14	3800	19	4	2	1	1	1	28	28	0.007 ± 0.001	0.007 ± 0.001
(AA)	2	400	2	0	1	0	0	1	4	4	0.010 ± 0.004	0.010 ± 0.004
IL10 (−819)
(CC)	40	8000	35	7	3	12	0	2	59	58	0.007 ± 0.001	0.007 ± 0.001
(CT)	10	2000	10	2	1	2	1	2	18	18	0.009 ± 0.001	0.009 ± 0.001
IL6
(GG)	32	6400	35	5	2	9	0	3	54	54	0.008 ± 0.001	0.008 ± 0.001
(GC)	18	3600	12	3	2	5	1	1	24	24	0.007 ± 0.002	0.007 ± 0.002
TNF
(GG)	33	6600	29	5	4	6	1	0	49	48	0.007 ± 0.001	0.007 ± 0.001
(GA)	17	3400	16	4	0	8	0	0	28	28	0.008 ± 0.001	0.008 ± 0.001
TGFβ1 codon 10
(CC)	13	2600	9	3	2	4	0	0	18	17	0.007 ± 0.002	0.007 ± 0.002
(CT)	25	5000	27	3	1	8	0	4	43	43	0.009 ± 0.001	0.009 ± 0.001
(TT)	12	2400	9	3	1	2	1	0	16	16	0.007 ± 0.002	0.007 ± 0.002
TGFβ1 codon 25
(GG)	38	7600	37	6	1	11	0	3	58	57	0.008 ± 0.001	0.008 ± 0.001
(GC)	12	2400	8	3	3	3	1	1	19	19	0.008 ± 0.002	0.008 ± 0.002

N = Number of analyzed subjects; NSM = Number of scored metaphases; B’: chromatid breaks; B”: chromosome breaks; Dic = Dicentric chromosome; AF = Acentric fragments; R = Rings; TR = tri- or tetra-radials; CAs = Chromosome aberrations; CAB = cells with aberrations.

Perrey C, Turner SJ, Pravica V, Howell WM, Hutchinson IV. 1999. ARMS-PCR methodologies to determine IL-10, TNFα, TNFβ, and TGFβ1 gene polymorphisms. Transplant. Immunol. 7:127–128.

 PubMed Web of Science ®Google Scholar

Zakharyan R, Petrek M, Arakelyan A, Mrazek F, Atshemyan S, Boyajyan A. 2012. IL-6 promoter polymorphism and plasma levels in patients with schizophrenia. Tissue Antigens 80:136–142.

 PubMed Web of Science ®Google Scholar