Investigation of high-mobility group box 1 variants with lymph node status and colorectal cancer risk

Table 1 Distribution of selected risk factors and demographic variables in colorectal cancer cases and controls

Variable	Cases (n = 1003)		Controls (n = 1303)		P value1
	n	%	n	%
Age (years)	61.10 ± 12.17		61.40 ± 9.61		0.496
Age (years)					0.605
< 61	451	44.97	600	46.05
≥ 61	552	55.03	703	53.95
Sex					0.867
Male	620	61.81	801	61.47
Female	383	38.19	502	38.53
Smoking status					0.002a
Never	744	74.18	1038	79.66
Ever	259	25.82	265	20.34
Alcohol use					< 0.001a
Never	829	82.65	1167	89.56
Ever	174	17.35	136	10.44
BMI (kg/m2)
< 24	670	66.80	688	52.80	< 0.001a
≥ 24	333	33.20	615	47.20
Site of tumor
Colon cancer	431	42.97
Rectum cancer	572	57.03

^aP < 0.05.

¹Two-sided χ² test and Student t-test.

BMI: Body mass index.

Table 2 Data for high-mobility group box 1 rs1412125 T > C and rs1045411 C > T nucleotide polymorphisms

Genotyped SNPs	Chromosome	Chr Pos (NCBI build 37)	Region	MAF for Chinese in database	MAF in our controls (n = 1303)	P value for HWE test in our controls	Genotyping method	Genotyping value (%)
HMGB1 rs1412125 T > C	13	31041595	5’flanking	0.24	0.24	0.064	SNPscan	98.87
HMGB1 rs1045411 C > T	13	31033232	3’UTR	0.20	0.20	0.862	SNPscan	98.87

SNPs: Single nucleotide polymorphisms; MAF: Minor allele frequency; HWE: Hardy-Weinberg equilibrium; 3’UTR: 3′untranslated region; HMGB1: High-mobility group box 1.

Table 3 Logistic regression analyses of associations between high-mobility group box 1 rs1412125 T > C polymorphisms and colorectal cancer risk

Genotype	Cases (n = 1003)		Controls (n = 1303)		Crude OR (95%CI)	P value	Adjusted OR (95%CI)1	P value
	n	%	n	%
HMGB1 rs1412125 T > C
TT	506	51.63	731	56.23	1.00		1.00
TC	395	40.31	504	38.77	1.13 (0.95-1.35)	0.161	1.14 (0.95-1.35)	0.160
CC	79	8.06	65	5.00	1.76 (1.24-2.49)	0.002	1.74 (1.22-2.47)	0.002
TC + CC	474	48.37	569	43.77	1.20 (1.02-1.42)	0.029	1.20 (1.02-1.43)	0.032
TT + TC	901	91.94	1235	95.00	1.00		1.00
CC	79	8.06	65	5.00	1.67 (1.19-2.34)	0.003	1.64 (1.17-2.32)	0.005
C allele	553	28.21	634	24.38
HMGB1 rs1045411 C > T
CC	627	63.98	823	63.31	1.00		1.00
CT	307	31.33	424	32.62	0.95 (0.79-1.14)	0.579	0.94 (0.78-1.13)	0.495
TT	46	4.69	53	4.08	1.14 (0.76-1.71)	0.532	1.10 (0.72-1.66)	0.664
CT + TT	353	36.02	477	36.69	0.97 (0.82-1.15)	0.742	0.96 (0.80-1.14)	0.615
CC + CT	934	95.31	1247	95.92	1.00		1.00
TT	46	4.69	53	4.08	1.16 (0.77-1.74)	0.475	1.12 (0.74-1.69)	0.589
T allele	399	20.36	530	20.38

¹Adjusted for age, sex, body mass index, smoking and drinking status.

OR: Odds ratio; CI: Confidence interval; HMGB1: High-mobility group box 1.

Table 4 Stratified analyses between high-mobility group box 1 rs1412125 T > C and colorectal cancer risk by sex, age, body mass index, smoking status and alcohol consumption

Variable	HMGB1 rs1412125 T > C (case/control)1			Adjusted OR (95% CI); P value2
	TT	TC	CC	Additive model	Homozygote model	Dominant model	Recessive model
Sex
Male	317/461	240/301	47/37	1.16 (0.93-1.45); 0.198	1.77 (1.12-2.81); 0.015	1.23 (0.99-1.53); 0.063	1.67 (1.06-2.62); 0.027
Female	189/270	155/203	32/28	1.11 (0.83-1.47); 0.494	1.77 (1.02-3.08); 0.043	1.18 (0.90-1.56); 0.230	1.69 (0.99-2.90); 0.055
Age
< 61	199/299	172/213	28/28	1.19 (0.92-1.55); 0.189	1.44 (0.85-2.44); 0.181	1.22 (0.95-1.57); 0.120	1.33 (0.79-2.22); 0.284
≥ 61	307/432	223/291	51/37	1.09 (0.86-1.39); 0.487	2.05 (1.27-3.31); 0.003	1.19 (0.95-1.50); 0.130	1.98 (1.24-3.16); 0.004
Smoking status
Never	371/573	297/409	60/53	1.11 (0.91-1.36); 0.306	1.73 (1.16-2.57); 0.007	1.18 (0.97-1.43); 0.090	1.65 (1.12-2.43); 0.012
Ever	135/158	98/95	19/12	1.21 (0.83-1.75); 0.320	1.76 (0.82-3.80); 0.148	1.27 (0.89-1.81); 0.185	1.64 (0.77-3.48); 0.201
Alcohol consumption
Never	416/650	327/457	67/57	1.11 (0.92-1.34); 0.285	1.80 (1.23-2.62); 0.002	1.19 (0.99-1.42); 0.067	1.72 (1.19-2.48); 0.004
Ever	90/81	68/47	12/8	1.29 (0.79-2.09); 0.310	1.37 (0.53-3.55); 0.522	1.30 (0.82-2.06); 0.271	1.24 (0.49-3.14); 0.657
BMI (kg/m2)
< 24	338/379	262/273	56/34	1.10 (0.88-1.38); 0.397	1.90 (1.21-3.00); 0.006	1.19 (0.96-1.48); 0.114	1.82 (1.17-2.84); 0.008
≥ 24	168/352	133/231	23/31	1.21 (0.91-1.60); 0.196	1.58 (0.89-2.81); 0.122	1.25 (0.95-1.64); 0.111	1.46 (0.83-2.56); 0.192

¹The genotyping was successful in 980 (97.71%) cases, and 1300 (99.77%) controls for HMGB1 rs1412125 T > C.

²Adjusted for age, sex, body mass index, smoking status and alcohol consumption (besides stratified factors accordingly) in a logistic regression model.

OR: Odds ratio; CI: Confidence interval; BMI: Body mass index; HMGB1: High-mobility group box 1.

Table 5 Stratified analyses between high-mobility group box 1 rs1045411 C > T polymorphism and colorectal cancer risk by sex, age, body mass index, smoking status and alcohol consumption

Variable	HMGB1 rs1045411 C > T (case/control)1			Adjusted OR (95% CI); P value2
	CC	CT	TT	Additive model	Homozygote model	Dominant model	Recessive model
Sex
Male	388/522	186/245	30/32	0.99 (0.78-1.26); 0.951	1.18 (0.70-1.99); 0.547	1.01 (0.81-1.27); 0.904	1.18 (0.70-1.98); 0.536
Female	239/301	121/179	16/21	0.87 (0.65-1.17); 0.366	1.03 (0.52-2.05); 0.927	0.89 (0.67-1.18); 0.418	1.08 (0.55-2.14); 0.817
Age
< 60	250/335	132/182	17/23	0.96 (0.73-1.26); 0.749	0.93 (0.51-1.72); 0.822	0.95 (0.74-1.24); 0.721	0.95 (0.52-1.73); 0.859
≥ 60	377/488	175/242	29/30	0.92 (0.72-1.18); 0.518	1.24 (0.70-2.21); 0.459	0.95 (0.75-1.21); 0.702	1.28 (0.72-2.25); 0.402
Smoking status
Never	472/647	226/343	30/45	0.89 (0.72-1.10); 0.285	0.92 (0.56-1.49); 0.720	0.90 (0.73-1.09); 0.278	0.95 (0.59-1.54); 0.837
Ever	155/176	81/81	16/8	1.12 (0.76-1.63); 0.576	2.08 (0.86-5.05); 0.107	1.20 (0.84-1.74); 0.320	2.00 (0.83-4.83); 0.121
Alcohol consumption
Never	522/736	250/382	38/46	0.92 (0.75-1.12); 0.397	1.15 (0.73-1.80); 0.547	0.94 (0.78-1.14); 0.544	1.18 (0.76-1.84); 0.463
Ever	105/87	57/42	8/7	1.12 (0.68-1.85); 0.650	0.81 (0.28-2.36); 0.692	1.08 (0.67-1.73); 0.767	0.77 (0.27-2.24); 0.635
BMI (kg/m2)
< 24	413/433	212/221	31/32	1.00 (0.79-1.26); 0.994	1.00 (0.60-1.68); 0.990	1.00 (0.80-1.25); 0.998	1.00 (0.60-1.67); 0.989
≥ 24	214/390	95/203	15/21	0.86 (0.64-1.17); 0.338	1.38 (0.69-2.75); 0.357	0.91 (0.69-1.21); 0.523	1.45 (0.73-2.87); 0.284

¹The genotyping was successful in 980 (97.71%) cases, and 1300 (99.77%) controls for HMGB1 rs1045411 C > T.

²Adjusted for age, sex, body mass index, smoking status and alcohol consumption (besides stratified factors accordingly) in a logistic regression model.

OR: Odds ratio; CI: Confidence interval; BMI: Body mass index; HMGB1: High-mobility group box 1.

Table 6 Stratified analyses between high-mobility group box 1 polymorphisms and colorectal cancer risk by site of tumor

Genotype	Controls (n=1303)		Colon cancer cases (n = 431)		Crude OR (95%CI)	P value	Adjusted OR (95%CI)1	P value1	Rectum cancer cases (n = 572)		Crude OR (95%CI)	P value	Adjusted OR (95%CI)1	P value1
	n	%	n	%					n	%
HMGB1 rs1412125 T > C
TT	731	56.23	215	50.83	1.00		1.00		291	52.24	1.00		1.00
TC	504	38.77	173	40.90	1.17 (0.93-1.47)	0.188	1.16 (0.92-1.46)	0.212	222	39.86	1.11 (0.90-1.36)	0.341	1.12 (0.91-1.39)	0.287
CC	65	5.00	35	8.27	1.83 (1.18-2.84)	0.007	1.81 (1.16-2.82)	0.009	44	7.90	1.70 (1.13-2.55)	0.010	1.69 (1.11-2.55)	0.014
TC + CC	569	43.77	208	49.17	1.24 (1.00-1.55)	0.053	1.23 (0.99-1.54)	0.064	266	47.76	1.17 (0.96-1.43)	0.114	1.19 (0.97-1.46)	0.097
TT + TC	1235	95.00	388	91.73	1.00		1.00		513	92.10	1.00		1.00
CC	65	5.00	35	8.27	1.72 (1.12-2.63)	0.013	1.70 (1.10-2.61)	0.016	44	7.90	1.63 (1.10-2.42)	0.016	1.61 (1.07-2.40)	0.022
C allele	634	24.38	243	28.72					310	27.83
HMGB1 rs1045411 C > T
CC	627	63.98	279	65.96	1.00		1.00		348	62.48	1.00		1.00
CT	307	31.33	123	29.08	0.86 (0.67-1.09)	0.208	0.85 (0.67-1.09)	0.200	184	33.03	1.03 (0.83-1.27)	0.812	1.01 (0.82-1.26)	0.900
TT	46	4.69	21	4.96	1.17 (0.69-1.97)	0.559	1.12 (0.66-1.90)	0.673	25	4.49	1.12 (0.68-1.82)	0.663	1.08 (0.65-1.78)	0.776
CT + TT	353	36.02	144	34.04	0.89 (0.71-1.12)	0.324	0.88 (0.70-1.12)	0.294	209	37.52	1.04 (0.84-1.27)	0.734	1.02 (0.83-1.26)	0.846
CC + CT	934	95.31	402	95.04	1.00		1.00		532	95.51	1.00		1.00
TT	46	4.69	21	4.46	1.23 (0.73-2.06)	0.435	1.18 (0.70-1.99)	0.535	25	4.49	1.11 (0.68-1.80)	0.686	1.07 (0.65-1.76)	0.788
T allele	399	20.36	165	19.50					234	21.01

¹Adjusted for age, sex, smoking status, alcohol use and body mass index status.

OR: Odds ratio; CI: Confidence interval; HMGB1: High-mobility group box 1.

Table 7 Logistic regression analyses of association between high-mobility group box 1 single nucleotide polymorphisms and lymph node status of colorectal cancer patients

Genotype	Positive (n = 518)		Negative (n = 485)		Crude OR (95%CI)	P value	Adjusted OR (95%CI)1	P value
	n	%	n	%
HMGB1 rs1412125 T > C
TT	259	50.88	247	52.44	1.00		1.00
TC	208	40.86	187	39.70	1.06 (0.82-1.38)	0.661	1.05 (0.81-1.37)	0.700
CC	42	8.25	37	7.86	1.08 (0.67-1.74)	0.744	1.09 (0.68-1.76)	0.712
TC + CC	250	49.12	224	47.56	1.06 (0.83-1.37)	0.626	1.06 (0.82-1.36)	0.650
TT + TC	467	91.75	434	92.14	1.00		1.00
CC	42	8.25	37	7.86	1.06 (0.67-1.67)	0.820	1.07 (0.67-1.70)	0.776
C allele	292	28.68	261	27.71
HMGB1 rs1045411 C > T
CC	315	61.89	312	66.24	1.00		1.00
CT	169	33.20	138	29.30	1.21 (0.92-1.60)	0.167	1.21 (0.92-1.59)	0.182
TT	25	4.91	21	4.46	1.18 (0.65-2.15)	0.591	1.19 (0.65-2.17)	0.578
CT + TT	194	38.11	159	33.76	1.21 (0.93-1.57)	0.156	1.20 (0.93-1.57)	0.167
CC + CT	484	95.09	450	95.54	1.00		1.00
TT	25	4.91	21	4.46	1.11 (0.61-2.01)	0.738	1.12 (0.62-2.03)	0.718
T allele	219	21.51	180	19.11

¹Adjusted for age, sex, alcohol use and smoking status.

OR: Odds ratio; CI: Confidence interval; HMGB1: High-mobility group box 1.

Table 8 Logistic regression analyses of association between high-mobility group box 1 single nucleotide polymorphisms and lymph node status of colorectal cancer in different region

Genotype	Colon cancer								Rectum cancer
	Positive (n = 205)		Negative (n = 226)		Crude OR (95%CI)	P value	Adjusted OR (95%CI)1	P value1	Positive (n = 313)		Negative (n = 259)		Crude OR (95%CI)	P value	Adjusted OR (95%CI)1	P value1
	n	%	n	%					n	%	n	%
HMGB1 rs1412125 T > C
TT	95	47.03	120	54.30	1.00		1.00		164	53.42	127	50.80	1.00		1.00
TC	87	43.07	86	38.91	1.28 (0.86-1.91)	0.232	1.29 (0.86-1.93)	0.216	121	39.41	101	40.40	0.93 (0.65-1.32)	0.676	0.92 (0.65-1.32)	0.657
CC	20	9.90	15	6.79	1.68 (0.82-3.47)	0.157	1.69 (0.82-3.48)	0.157	22	7.17	22	8.80	0.77 (0.41-1.46)	0.430	0.78 (0.41-1.49)	0.455
TC + CC	107	52.97	101	45.70	1.34 (0.91-1.96)	0.136	1.35 (0.92-1.98)	0.126	143	46.58	123	49.20	0.90 (0.64-1.26)	0.538	0.90 (0.64-1.26)	0.531
TT + TC	182	90.10	206	93.21	1.00		1.00		285	92.83	228	91.20	1.00		1.00
CC	20	9.90	15	6.79	1.51 (0.75-3.04)	0.248	1.51 (0.75-3.03)	0.252	22	7.17	22	8.80	0.80 (0.43-1.48)	0.478	0.81 (0.44-1.51)	0.509
C allele	127	31.44	116	26.24					165	26.87	145	29.00
HMGB1 rs1045411 C > T
CC	127	62.87	152	68.78	1.00		1.00		188	61.24	160	64.00	1.00		1.00
CT	66	32.67	57	25.79	1.39 (0.91-2.12)	0.133	1.41 (0.92-2.16)	0.117	103	33.55	81	32.40	1.08 (0.76-1.55)	0.667	1.08 (0.75-1.55)	0.685
TT	9	4.46	12	5.43	0.90 (0.37-2.20)	0.813	0.90 (0.37-2.21)	0.814	16	5.21	9	3.60	1.51 (0.65-3.52)	0.336	1.57 (0.67-3.68)	0.297
CT + TT	75	37.13	69	31.22	1.30 (0.87-1.95)	0.201	1.32 (0.88-1.98)	0.182	119	38.76	90	36.00	1.13 (0.80-1.59)	0.503	1.13 (0.80-1.60)	0.503
CC + CT	193	95.54	209	94.57	1.00		1.00		291	94.79	241	96.40	1.00		1.00
TT	9	4.46	12	5.43	0.81 (0.34-1.97)	0.645	0.81 (0.33-1.97)	0.644	16	5.21	9	3.60	1.47 (0.64-3.39)	0.364	1.53 (0.66-3.55)	0.320
T allele	84	20.79	81	18.33					135	21.99	99	19.80

¹Adjusted for age, sex, smoking status, alcohol use and body mass index status.

OR: Odds ratio; CI: Confidence interval; HMGB1: High-mobility group box 1.