Vitamin D receptor gene polymorphisms and hepatocellular carcinoma in alcoholic cirrhosis

Table 1 Main demographic and clinical characteristics of liver transplant recipients (n = 240) according to the presence (n = 80, 33.3%) or absence (n = 160, 66.7%) of HCC

		HCC present (n = 80)	HCC absent (n = 160)	P value
Male gender	178 (74.2)	70	108	< 0.0010
Age1 (yr)	55 (22-68)	57.2 ± 5.9	52.0 ± 8.6	< 0.0001
Body mass index1 (kg/m2)	25.2 (14.8-48.5)	26.4 ± 4.2	24.7 ± 3.3	< 0.0010
Etiology
Viral (n = 137)		50	87	NS
HBV	37 (15.4)
HCV	100 (41.7)
Alcoholic	103 (42.9)
Child-Pugh score1	8 (5-14)	7 (5-14)	8 (5-13)	< 0.0001
Child-Pugh score1 > 8 (n = 87)		22	65	< 0.0500
Diabetes mellitus	67 (27.9)	29	38	< 0.0500

The continuous variables are reported as means (standard deviation), whereas the categorical variables are reported as frequencies (%). Statistical analysis was performed using the Student’s t-test for continuous variables and the Pearson χ² test for categorical variables.

¹At transplant operation. HCC: Hepatocellular carcinoma; HBV: Hepatitis B virus; HCV: Hepatitis C virus; NS: Not significant.

Table 2 Allelic and genotypic frequencies of the VDR FokI C>T (F/f), BsmI A>G (B/b), ApaI T>G (A/a) and TaqI T>C (T/t) polymorphisms in controls and liver cirrhosis patients n (%)

VDR	Control subjects (n = 236)	Liver cirrhosis (n = 240)	OR	95% CI	P1
	F = 0.646	F = 0.665	1	Ref.
FokI	f = 0.354	f = 0.335	1.085	0.831-1.417	NS
	B = 0.392	B = 0.421	1	Ref.
BsmI	b = 0.608	b = 0.579	1.127	0.870-1.460	NS
	A = 0.570	A = 0.590	1	Ref.
ApaI	a = 0.430	a = 0.410	1.084	0.838-1.402	NS
	T = 0.608	T = 0.579	1	Ref.
TaqI	t = 0.392	t = 0.421	0.887	0.685-1.149	NS
	F/F = 104 (44.1)	F/F = 105 (43.8)	1	Ref.
FokI	F/f = 97 (41.1)	F/f = 109 (45.4)	1.113	0.758-1.635	NS
	f/f = 35 (14.8)	f/f = 26 (10.8)	0.736	0.416-1.303	NS
	B/B = 41 (17.4)	B/B = 40 (16.7)	1	Ref.
BsmI	B/b = 103 (43.6)	B/b = 122 (50.8)	1.214	0.732-2.014	NS
	b/b = 92 (39.0)	b/b = 78 (32.5)	0.869	0.513-1.473	NS
	A/A = 76 (32.2)	A/A = 80 (33.3)	1	Ref.
ApaI	A/a = 117 (49.6)	A/a = 123 (51.3)	0.999	0.668-1.494	NS
	a/a = 43 (18.2)	a/a = 37 (15.4)	0.817	0.477-1.400	NS
	T/T = 89 (37.7)	T/T = 76 (31.7)	1	Ref.
TaqI	T/t = 109 (46.2)	T/t = 126 (52.5)	1.354	0.909-2.017	NS
	t/t = 38 (16.1)	t/t = 38 (15.8)	1.171	0.681-2.013	NS

The odds ratios were constructed with the wild type for each polymorphism as the reference. The statistical analysis was carried out using the Pearson χ² test.

¹Pearson χ² test; OR: Odds ratio; CI: Confidence interval; VDR: Vitamin D receptor.

Table 3 Estimated VDR haplotype frequencies [FokI C>T (F/f), BsmI A>G (B/b), ApaI T>G (A/a) and TaqI T>C (T/t) polymorphisms] in control subjects and in patients with liver cirrhosis of viral and alcoholic origin

Control subjects (n = 236)			Alcoholic cirrhosis (n = 103)			Viral cirrhosis (n = 137)
Haplotypes	n	Frequencies	Haplotypes	n	Frequencies	Haplotypes	n	Frequencies
C-A-T-C	139	0.295	C-A-T-C	59	0.286	C-A-T-C	103	0.376
T-G-G-T	102	0.216	T-G-G-T	46	0.223	T-G-G-T	56	0.204
C-G-G-T	99	0.210	C-G-G-T	50	0.243	C-G-G-T	43	0.158
C-G-T-T	58	0.123	C-G-T-T	29	0.141	C-G-T-T	25	0.091
T-A-T-C	41	0.087	T-A-T-C	11	0.053	T-A-T-C	23	0.084
T-G-T-T	23	0.049	T-G-T-T	6	0.029	T-G-T-T	19	0.069
C-A-T-T	4	0.008	C-A-T-T	1	0.005	C-A-T-T	3	0.011
C-G-T-C	3	0.006	C-G-T-C	2	0.010	C-G-T-C	2	0.007
C-G-G-C	2	0.004	C-A-G-C	2	0.010
T-A-T-T	1	0.002

The statistical analysis was performed by means of the exact test for sample differentiation based on haplotype frequencies. Alcoholic vs viral liver cirrhosis P < 0.05.

Table 4 Genotypic frequencies of the VDR FokI C>T (F/f), BsmI A>G (B/b), ApaI T>G (A/a) and TaqI T>C (T/t) polymorphisms in liver cirrhosis patients grouped according to the presence or absence of HCC

VDR	HCC absent (n = 160)	HCC present (n = 80)	OR	95% CI	P1
	F/F = 69 (43.1)	F/F = 36 (45.0)	1	Ref.
FokI	F/f = 73 (45.6)	F/f = 36 (45.0)	0.945	0.537-1.663	NS
	f/f = 18 (11.3)	f/f = 8 (10.0)	0.852	0.345-2.113	NS
	B/B = 28 (17.5)	B/B = 12 (15.0)	1	Ref.	2
BsmI	B/b = 87 (54.4)	B/b = 35 (43.8)	0.939	0.433-2.028	NS
	b/b = 45 (28.1)	b/b = 33 (41.2)	1.711	0.766-3.813	NS
	A/A = 53 (33.1)	A/A = 27 (33.8)	1	Ref.
ApaI	A/a = 85 (53.1)	A/a = 38 (47.5)	0.878	0.483-1.595	NS
	a/a = 22 (13.8)	a/a = 15 (18.7)	1.338	0.605-2.968	NS
	T/T = 44 (27.5)	T/T = 32 (40.0)	1	Ref.	3
TaqI	T/t = 88 (55.0)	T/t = 38 (47.5)	0.594	0.329-1.072	0.08
	t/t = 28 (17.5)	t/t = 10 (12.5)	0.491	0.212-1.141	0.09

The odds ratios were constructed with the wild type for each polymorphism as the reference. The statistical analysis was carried out using the Pearson χ² test.

¹Pearson χ² test;

²B/B + B/b genotypes vs b/b genotype: P < 0.05;

³T/t + t/t genotypes vs T/T genotype: P < 0.05.

Table 5 Estimated VDR haplotype frequencies [FokI C>T (F/f), BsmI A>G (B/b), ApaI T>G (A/a) and TaqI T>C (T/t) polymorphisms] in patients with liver cirrhosis of viral and alcoholic origin according to the presence of HCC

Alcoholic cirrhosis (n = 103)						Viral cirrhosis (n = 137)
HCC present (n = 30)			HCC absent (n = 73)			HCC present (n = 50)			HCC absent (n = 87)
Haplotypes	n	Frequencies	Haplotypes	n	Frequencies	Haplotypes	n	Frequencies	Haplotypes	n	Frequencies
C-A-T-C	9	0.150	C-A-T-C	50	0.343	C-A-T-C	40	0.40	C-A-T-C	64	0.368
T-G-G-T	16	0.267	T-G-G-T	30	0.205	T-G-G-T	15	0.15	T-G-G-T	38	0.218
C-G-G-T	18	0.300	C-G-G-T	32	0.219	C-G-G-T	19	0.19	C-G-G-T	27	0.155
C-G-T-T	14	0.233	C-G-T-T	15	0.103	C-G-T-T	5	0.05	C-G-T-T	17	0.098
T-A-T-C	1	0.017	T-A-T-C	10	0.068	T-A-T-C	7	0.07	T-A-T-C	15	0.086
T-G-T-T	2	0.033	T-G-T-T	4	0.027	T-G-T-T	11	0.11	T-G-T-T	11	0.063
			C-A-T-T	1	0.007	C-A-T-T	2	0.02	C-G-T-C	1	0.006
			C-G-T-C	2	0.014	C-G-T-C	1	0.01	T-A-T-T	1	0.006
			C-G-G-C	2	0.014

The statistical analysis was performed by means of the exact test for sample differentiation based on haplotype frequencies. Non-viral liver cirrhosis with HCC vs (a) viral liver cirrhosis without HCC, P < 0.01, (b) viral liver cirrhosis with HCC, P < 0.05.