Association of hydroxysteroid 11-beta dehydrogenase 1 polymorphisms with chronic liver fibrosis and the occurrence of hepatocellular carcinoma in a Han Chinese population

Figures & data

Figure 1. The location of the HSD11B1 gene and six selected SNPs.

Table 1. Primer sequences and reaction conditions for genotyping HSD11B1 polymorphisms.

SNPs	Sequences of the primers	Annealing temperature (°C)
Rs846908	R:5′-ACGTTGGATGTGCAAGCAAAAGTGTCATTC-3′	46.8
	R:5′-ACGTTGGATGGGAGAAACACCATTAGACAG-3′
Rs701950	R:5′-ACGTTGGATGCAGTGCCTAGCATAGAGTAG-3′	46.8
	R:5′-ACGTTGGATGGGACTACAGCATGTCAAAAC-3′
Rs846910	R:5′-ACGTTGGATGCACTCTGTTGCTTGTGCTTG-3′	45.6
	R:5′-ACGTTGGATGAGGTGAGTATTAGAGCCCAG-3′
Rs3753519	R:5′-ACGTTGGATGATCCCATCCCCCAAATCTTC-3′	46.2
	R:5′-ACGTTGGATGAATGCTAGCTGAAACAGTTG-3′
Rs4844488	R:5′-ACGTTGGATGGAGAAGGCTTACAACAAATAC-3′	48.9
	R:5′-ACGTTGGATGACTGAGTCTAATTTGGGCAC-3′
Rs932335	R:5′-ACGTTGGATGTACAAAGCCTACTACAGCTC-3′	49.0
	R:5′-ACGTTGGATGCCACTTTCCCTAAATGACAG-3′

Table 2. General and clinical characteristics of study subjects.

Characteristics	CHB (N=104)	P-valuea	LC (N=266)	P-valueb	HCC (N=286)	Non-HCC (N=370)	P-valuec	Reference ranges
Sex (M/F)	84/20	0.756	211/55	0.050	245/41	295/75	0.048
Age, M (P25, P75)	47(43,53)	0.340	49(41,56)	0.144	50(46,56)	49(42,55)	0.022
Smoking, n (%)		0.804		0.000			0.000
Have ever smoked	37(35.6)		91(34.2)		144(50.3)	128(34.6)
Have never smoked	67(64.4)		175(65.8)		142(49.7)	242(65.4)
Alcohol consumption, n (%)		0.122		0.131			0.023
Have ever consumed alcohol	28(26.9)		94(35.3)		119(41.6)	122(33.0)
Have never consumed alcohol	78(73.1)		172(64.7)		167(58.4)	248(67.0)
Serum HBV-DNA, n (%)		0.000		0.006			0.118
Positive	99(96.1)		213(81.0)		238(89.5)	312(85.2)
Negative	4(3.9)		50(19.0)		28(10.5)	54(14.8)
HBV load, log10 (IU/mL)	6.2(4.4,7.4)	0.001	5.4(3.6,6.5)	0.034	5.0(3.4,6.0)	5.64(4.0,6.9)	0.000	1.3-8.2
HBeAg, n (%)		0.347		0.545			0.775
Positive	49(47.1)		111(41.7)		100(44.4)	160(43.2)
Negative	55(52.9)		155(58.3)		125(55.6)	210(56.8)
ALT (U/L), M (P25, P75)	172(46,494)	0.000	41(24,83)	0.600	43(27,69)	51(26,157)	0.005	13.0-35.0
AST (U/L), M (P25, P75)	98(40,246)	0.000	47(31,90)	0.000	62(38,110)	51(33,122)	0.087	7.0-40.0
ALP (U/L), M (P25, P75)	87(67,123)	0.633	89(68,125)	0.000	130(86,191)	89(68,124)	0.000	50.0-135.0
GGT (U/L), M (P25, P75)	93(38,162)	0.000	27(50,100)	0.000	113(54,254)	55(29,128)	0.000	7.0-45.0
Prealbumin (g/L), M (P25, P75)	0.16(0.13,0.20)	0.000	0.12(0.09,0.16)	0.179	0.13(0.08,0.17)	0.13(0.09,0.17)	0.695	0.18-0.39
Albumin (g/L), M (P25, P75)	36(32,41)	0.000	31(25,36)	0.000	33(28,37)	32(27,38)	0.328	40.0-55.0
Total bilirubin (umol/L), M (P25, P75)	20(13,48)	0.026	26(17,44)	0.274	26(17,44)	25(15,60)	0.866	0.0-8.6
Cholinesterase (U/L), M (P25, P75)	6019(4343,8281)	0.000	3286(2338,4743)	0.008	3918(2488,5762)	3893(2654,5804)	0.584	4300–12000
Platelet count (x109/L), M (P25,P75)	145(117,188)	0.000	28(16,61)	0.000	119(79,172)	99(58,145)	0.000	100–300

^a CHB vs. LC; ^bLC vs. HCC; ^cNon-HCC (included CHB and LC patients) vs. HCC, calculated by analysis of variance and the least significant difference multiple comparisons test. CHB, chronic hepatitis B; LC, liver cirrhosis; HCC, hepatocellular carcinoma; Non-HCC, Non-chronic HBV infection HCC; ALT, alanine transaminase; AST, aspartate transaminase; ALP, alkaline; phosphatase; GGT, glutamyl transpeptidase; M, male; F, female; HBeAg, hepatitis B e antigen.

Table 3. The Hardy-Weinberg equilibrium of each SNP in each group.

SNP	CHB (N=104)	LC (N=266)	HCC (N=286)	Non-HCC (N=370)
rs846908	0.596	0.258	0.615	0.288
rs701950	0.334	0.474	0.675	0.900
rs846910	0.996	0.251	0.799	0.334
rs3753519	0.426	0.634	0.955	0.978
rs4844488	0.831	0.218	0.762	0.353
rs932335	0.173	0.001	0.227	0.025

Note: CHB, chronic hepatitis B; LC, liver cirrhosis; HCC, hepatocellular carcinoma; Non-HCC, Non-chronic HBV infection HCC.

Table 4. Genotype and allele frequencies of SNPs in the HSD11B1 gene in each group.

			CHB vs. LC			LC vs. HCC			Non-HCC vs. HCC
	CHB	LC	OR(95%Cl) ^a	P-value^a	HCC	OR(95%Cl)^b	P-value^b	Non-HCC	OR(95%Cl)^c	P-value^c
rs846908
Detected number	(N=104)	(N=266)			(N=284)			(N=370)
AA	7(6.7)	24(9.0)	1		22(7.7)	1		31(8.4)	1
AG	44(42.3)	99(37.2)	0.65(0.26-1.64)	0.358	108(38.0)	1.15(0.60-2.21)	0.669	143(38.6)	1.02(0.55-1.87)	0.959
GG	53(51.0)	143(53.8)	0.76(0.31-1.88)	0.550	154(54.2)	1.11(0.59-2.09)	0.752	196(53.0)	1.05(0.58-1.90)	0.876
^#GG + AG	97	242	0.71(0.29-1.72)	0.448	116	1.13(0.61-2.08)	0.706	339	1.04(0.58-1.85)	0.908
*AA + AG	51	123	1.09 (0.69-1.72)	0.713	130	1.00(0.71-1.41)	1.00	174	1.06(0.77-1.45)	0.731
PIC	0.402	0.400			0.392			0.401
A Allele	58(27.9)	147(27.6)	1		152(26.8)	1		205(27.7)	1
G Allele	150(72.1)	385(72.4)	1.01(0.71-1.45)	0.945	416(73.2)	1.05(0.80-1.36)	0.746	535(72.3)	1.05(0.82-1.34)	0.705
rs701950
Detected number	(N=104)	(N=266)			(N=282)			(N=370)
GG	2(4.1)	12(4.5)	1		10(3.5)	1		14(3.8)	1
GA	34(32.7)	80(30.1)	0.39(0.08-1.84)	0.234	92(32.6)	1.21(0.49-2.99)	0.683	114(30.8)	1.50(0.44-2.50)	0.916
AA	68(65.4)	174(65.4)	0.41(0.09-1.91)	0.258	180(63.8)	1.07(0.44-2.58)	0.873	242(65.4)	0.95(0.41-2.22)	0.902
^#AA + GA	102	102	0.41(0.09-1.86)	0.245	272	1.14(0.47-2.66)	0.808	356	0.98(0.42-2.27)	0.963
*GG + GA	36	92	0.98(0.61-1.58)	0.932	102	0.91(0.63-1.29)	0.584	128	0.91(0.63-1.29)	0.584
PIC	0.299	0.314			0.319			0.310
G Allele	38(18.3)	104(19.5)	1		112(19.9)	1		142(19.2)	1
A Allele	170(81.7)	428(80.5)	0.92(0.61-1.)	0.691	452(80.1)	0.98(0.73-1.32)	0.898	598(80.8)	0.96(0.73-1.26)	0.763
rs846910
Detected number	(N=104)	(N=264)			(N=285)			(N=368)
AA	7(6.7)	20(7.6)	1		19(6.7)	1		27(7.3)	1
AG	40(38.5)	92(34.8)	0.77(0.30-1.98)	0.586	106(37.2)	1.19(0.59-2.40)	0.621	132(35.9)	1.10(0.57-2.11)	0.776
GG	57(54.8)	152(57.6)	0.88(0.35-2.20)	0.781	160(56.1)	1.08(0.55-2.13)	0.825	209(56.8)	1.05(0.56-1.97)	0.884
^#GG + AG	97	244	0.83(0.34-2.05)	0.691	266	1.12(0.58-2.18)	0.733	341	1.07(0.58-1.98)	0.835
*AA + AG	47	112	1.09(0.69-1.73)	0.707	125	0.93(0.66-1.31)	0.686	159	0.97(0.71-1.33)	0.842
PIC	0.385	0.375			0.378			0.378
A Allele	54(26.0)	132(25.0)	1		144(25.3)			186(25.3)	1
G Allele	154(74.0)	396(75.0)	1.05(0.73-1.52)	0.787	426(74.7)	0.99(0.75-1.30)	0.920	550(74.7)	1.00(0.78-1.29)	0.997
rs3753519
Detected number	(N=103)	(N=263)			(N=281)			(N=366)
TT	12(11.7)	23(8.7)	1		30(10.7)	1		35(9.6)	1
TC	41(39.8)	115(43.7)	1.44(0.66-3.19)	0.363	123(43.8)	0.73(0.43-1.46)	0.457	156(42.6)	0.90(0.52-1.57)	0.715
CC	50(48.5)	125(47.5)	1.27(0.58-2.76)	0.550	128(45.8)	0.74(0.40-1.36)	0.335	175(47.8)	0.81(0.47-1.40)	0.452
^#CC + TC	91	240	1.35(0.64-2.84)	0.433	251	0.77(0.43-1.37)	0.370	331	0.85(0.51-1.44)	0.554
*TT + TC	53	138	0.94(0.60-1.49)	0.804	153	0.90(0.64-1.26)	0.532	191	0.90(0.64-1.26)	0.535
PIC	0.432	0.425			0.439			0.427
T Allele	65(31.6)	161(30.6)	1		183(32.6)	1		226(30.9)	1
C Allele	141(68.4)	365(69.4)	1.05(0.74-1.48)	0.803	379(67.4)	0.91(0.71-1.18)	0.489	506(69.1)	0.93(0.73-1.17)	0.517
rs4844488
Detected number	(N=104)	(N=265)			(N=283)			(N=369)
AA	48(46.2)	141(53.2)	1		141(49.8)	1		189(51.2)	1
AG	46(44.2)	99(37.4)	0.76(0.46-1.23)	0.258	119(42.0)	1.22(0.85-1.75)	0.278	145(39.3)	1.13(0.81-1.57)	0.485
GG	10(9.6)	25(9.4)	0.90(0.40-2.02)	0.798	23(8.1)	0.92(0.49-1.71)	0.791	35(9.5)	0.9(0.51-1.61)	0.721
^#AA + AG	94	240	1.02(0.47-2.22)	0.957	260	0.84(0.46-1.54)	0.577	334	0.84(0.46-1.54)	0.577
*GG + AG	56	124	0.78(0.49-1.24)	0.294	142	1.16(0.82-1.63)	0.396	180	1.08(0.79-1.48)	0.626
PIC	0.433	0.404			0.413			0.413
A Allele	142(68.3)	381(71.9)	1		401(70.8)	1		523(70.9)	1
G Allele	66(31.7)	149(28.1)	0.84(0.59-1.19)	0.331	165(29.2)	1.05(0.81-1.37)	0.704	215(29.1)	1.00(0.79-1.27)	0.994
rs932335
Detected number	(N=104)	(N=266)			(N=285)			(N=370)
GG	70(67.3)	178(66.9)	1		204(71.6)	1		246(66.5)	1
GC	33(31.7)	68(25.6)	0.82(0.49-1.35)	0.427	71(24.9)	0.92(0.62-1.36)	0.662	103(27.8)	0.86(0.60-1.24)	0.429
CC	1(1.0)	20(7.5)	7.89(1.04-60.06)	0.046	10(3.5)	0.47(0.21-1.04)	0.062	21(5.7)	0.61(0.28-1.34)	0.218
^#GG + GC	103	246	8.37(1.11-63.39)	0.04	275	0.48(0.22-1.06)	0.068	349	0.63(0.29-1.39)	0.254
*CC + GC	34	88	1.03(0.63-1.67)	0.916	81	0.82(0.57-1.18)	0.281	124	0.82(0.58-1.16)	0.259
PIC	0.273	0.324			0.269			0.310
G Allele	174(83.7)	424(79.7)	1		479(84.0)	1		598(80.8)	1
C Allele	34(16.3)	108(20.3)	1.37(0.89-2.08)	0.148	91(16.0)	0.75(0.55-1.02)	0.062	142(19.2)	0.80(0.60-1.07)	0.130

P-value^a,b,c represents the CHB vs. LC, LC vs. HCC, Non-HCC (included CHB and LC patients) vs. HCC, the results are adjusted for age, gender, smoking, and drinking by logistic. CHB, chronic hepatitis B; LC, liver cirrhosis; HCC, hepatocellular carcinoma; Non-HCC, Non-chronic HBV infection HCC; PIC, polymorphic information content. # represent recessive model analysis; * represent dominant model analysis.

Table 5. The impact of rs932335 genotypes on the clinical indicators of liver cirrhosis.

Characteristics	GG (N=178)	GC (N=68)	CC (N=20)	P-value	Reference ranges
Sex (M/F)	143/35	52/16	16/4	0.797
Age, M (P25, P75)	50(42,58)	47(39,54)	50(42,59)	0.325
Smoking, n (%)				0.648
Have ever smoked	63(35.4)	23(33.8)	5(25.0)
Have never smoked	115(64.6)	45(66.2)	15(75.0)
Alcohol consumption, n (%)				0.196
Have ever consumed alcohol	69(38.8)	18(26.5)	7(35.0)
Have never consumed alcohol	109(61.2)	50(73.5)	13(65.0)
Serum HBV-DNA, n (%)				0.652
Positive	147(79.7)	56(84.8)	16(80.0)
Negative	36(20.3)	10(15.2)	4(20.0)
HBV load, log10 (IU/mL)	5.4(3.5,6.5)	5.4(4.0,7.0)	5.5(4.2,6.4)	0.764	1.3-8.2
HBeAg, n (%)				0.975
Positive	74(41.6)	29(42.6)	8(40.0)
Negative	104(58.4)	39(57.4)	12(60.0)
ALT (U/L), M (P25, P75)	37(21,98)	39(23,71)	57(29,161)	0.022	13.0-35.0
AST (U/L), M (P25, P75)	46(30,67)	40(30,91)	61(38,145)	0.010	7.0-40.0
ALP (U/L), M (P25, P75)	87(67,126)	97(71,130)	86(65,107)	0.975	50.0-135.0
GGT (U/L), M (P25, P75)	46(26,90)	63(30,119)	113(54,254)	0.466	7.0-45.0
Prealbumin (g/L), M (P25, P75)	0.12(0.09,0.16)	0.12(0.08,0.15)	0.12(0.10,0.13)	0.787	0.18-0.39
Albumin (g/L), M (P25,P75)	31(25,36)	30(25,33)	30(25,39)	0.405	40.0-55.0
Total bilirubin (umol/L), M (P25, P75)	27(16,60)	31(19,71)	22(15,45)	0.302	0.0-8.6
Cholinesterase (U/L), M (P25, P75)	3355(2297,4812)	2909(2349,4123)	4083(2559,5799)	0.222	4300–12000
Platelet count (×10^9/L), M (P25,P75)	77(53,120)	71(48,117)	81(63,189)	0.258	100–300
Child-Pugh score	9(6,11)	9(7,11)	8.5(7.0,10.7)	0.455	1–15
Child-Pugh level				0.557
A	1(0.6)	0(0.0)	0(0.0)
B	43(24.2)	10(14.7)	4(20.0)
C	61(34.3)	28(41.2)	10(50.0)
D	73(41.0)	30(44.1)	6(30.0)
MELD score	10(8,15)	10(7,12)	12(9,17)	0.022

Note: M/F, male/female; HBV, hepatitis B virus; HBeAg, HBV e antigen; ALT, alanine transaminase; AST, aspartate transaminase; ALP, alkaline; phosphatase; GGT, glutamyl transpeptidase; CHB, chronic hepatitis B; LC, liver cirrhosis; HCC, hepatocellular carcinoma; Non-HCC, Non-chronic HBV infection HCC.

Table 6 Genotype, allele frequency and genetic diversity of six SNP locus of HSD11B1.

SNP		Genotype frequency			Allele frequency				P-value
rs846908		AA	GA	GG	A	G	HO	HE
	CHB	7	44	53	58	150	0.423	0.402	0.8071
	LC	24	99	143	147	385	0.3722	0.3999	0.2825
	HCC	22	108	154	152	416	0.3803	0.392	0.6495
rs701950		AA	GA	GG	A	G	HO	HE
	CHB	68	34	2	170	38	0.327	0.299	0.5139
	LC	174	80	12	428	104	0.3008	0.3145	0.4402
	HCC	180	92	10	452	112	0.3262	0.3183	0.8514
rs846910		AA	AG	GG	A	G	HO	HE
	CHB	7	40	57	54	154	0.3846	0.3844	1.0000
	LC	20	92	152	132	396	0.3485	0.375	0.2519
	HCC	19	106	160	144	426	0.3719	0.3776	0.7560
rs3753519		TT	TC	CC	T	C	HO	HE
	CHB	12	41	50	65	141	0.3981	0.4319	0.4927
	LC	23	115	125	161	365	0.4373	0.4248	0.7714
	HCC	30	123	128	183	379	0.4377	0.4392	1.0000
rs4844488		GG	AG	AA	G	A	HO	HE
	CHB	10	46	48	66	142	0.4423	0.4332	1.0000
	LC	25	99	141	149	381	0.3736	0.4042	0.2247
	HCC	23	119	141	165	401	0.4205	0.4131	0.8855
rs932335		CC	CG	GG	C	G	HO	HE
	CHB	1	33	70	34	174	0.3269	0.2735	0.06722
	LC	20	68	178	108	424	0.2556	0.3236	0.001071
	HCC	10	71	204	91	479	0.2491	0.2683	0.2657

Note: CHB, chronic hepatitis B; LC, liver cirrhosis; HCC, hepatocellular carcinoma; HO, observed heterozygosity; HE, expected heterozygosity.

Data availability statement

Due to the nature of this research, the participants of this study did not agree to their data being shared publicly, so supporting data is not available.