Polymorphisms of folate metabolism genes in patients with cirrhosis and hepatocellular carcinoma

Table 1 Relationship between risk factors and hepatocellular carcinoma and liver cirrhosis development

Variables	Controls n (%)	Cirrhosis n (%)	1OR (95%CI)	P-value	HCC n (%)	OR (95%CI)	2P
Age
< 46 anos	238 (67)	22 (19)	10.31 (5.66-18.76)	P < 0.001	8 (11)	16.36 (6.68-40.05)	P < 0.001
≥ 46 anos	118 (33)	94 (81)			63 (89)
Genre
Female	95 (26.7)	30 (25.9)	0.96 (0.54-1.72)	P = 0.893	19 (26.8)	0.59 (0.29-1.22)	P = 0.154
Male	261 (73.3)	86 (74.1)			52 (73.2)
Alcoholic habit
Not	191 (54)	54 (46.6))	1.55 (0.91-2.63)	P = 0.106	27 (38)	2.01 (1.03-3.89)	P = 0.039
Yes	165 (46)	62 (53.4)			44 (62)
Smoking habit
Nonsmokers	199 (56)	72 (62)	0.47 (0.28-0.78)	P = 0.003	31 (43.7)	0.9 (0.48-1.67)	P = 0.734
Smokers	157 (44)	44 (38)			40 (56.3)

¹Odds ratio (OR) adjusted for age, genre, alcohol consumption, smoking habits and polymorphisms;

²P values significant at P ≤ 0.05. HCC: Hepatocellular carcinoma.

Table 2 Association of MTHFR C677T, MTHFR A1298C, MTR A2756G and MTRR A66G polymorphisms with hepatocellular carcinoma, adjusted for gender, age, smoking and alcohol consumption

Model	Genotipe	Controln (%)	Case n (%)	1OR (95%CI)	2P-value	Genotipe	Controln (%)	Case n (%)	1OR (95%CI)	2P-value
	MTHFR C677T					MTHFR A1298C
Codominant	C/C	149 (41.9)	28 (39.4)	1		A/A	205 (57.6)	32 (45.1)	1
	C/T	174 (48.9)	36 (50.7)	0.93 (0.51-1.68)	0.91	A/C	116 (32.6)	24 (33.8)	1.29 (0.69-2.42)	0.014
	T/T	33 (9.3)	7 (9.9)	1.13 (0.41-3.09)		C/C	35 (9.8)	15 (21.1)	3.37 (1.52-7.50)
Dominant	C/C	149 (41.9)	28 (39.4)	1		A/A	205 (57.6)	32 (45.1)	1
	C/T-T/T	207 (58.1)	43 (60.6)	0.96 (0.54-1.70)	0.88	A/C-C/C	151 (42.4)	39 (54.9)	1.71 (0.98-2.99)	0.06
Recessive	C/C-C/T	323 (90.7)	64 (90.1)	1		A/A-A/C	321 (90.2)	56 (78.9)	1
	T/T	33 (9.3)	7 (9.9)	1.18 (0.46-3.05)	0.73	C/C	35 (9.8)	15 (21.1)	3.04 (1.43-6.47)	0.0051
Overdominant	C/C-T/T	182 (51.1)	35 (49.3)	1		A/A-C/C	240 (67.4)	47 (66.2)	1
	C/T	174 (48.9)	36 (50.7)	0.91 (0.52-1.59)	0.73	A/C	116 (32.6)	24 (33.8)	0.99 (0.55-1.78)	0.98
Aditive	---	---	---	1.01 (0.64-1.58)	0.97	---	---	---	1.71 (1.16-2.52)	0.007
	MTR A2756G					MTRR A66G
Codominant	A/A	263 (73.9)	46 (64.8)	1		A/A	105 (29.5)	12 (16.9)	1
	A/G	83 (23.3)	21 (29.6)	1.58 (0.84-2.98)	0.13	A/G	179 (50.3)	50 (70.4)	3.26 (1.54-6.87)	< 0.001
	G/G	10 (2.8)	4 (5.6)	3.29 (0.81-13.30)		G/G	72 (20.2)	9 (12.7)	1.16 (0.44-3.11)
Dominant	A/A	263 (73.9)	46 (64.8)	1		A/A	105 (29.5)	12 (16.9)	1
	A/G-G/G	93 (26.1)	25 (35.2)	1.73 (0.95-3.15)	0.078	A/G-G/G	251 (70.5)	59 (83.1)	2.55 (1.24-5.25)	0.0072
Recessive	A/A-A/G	346 (97.2)	67 (94.4)	1		A/A-A/G	284 (79.8)	62 (87.3)	1
	G/G	10 (2.8)	4 (5.6)	2.91 (0.73-11.59)	0.15	G/G	72 (20.2)	9 (12.7)	0.51 (0.23-1.12)	0.077
Overdominant	A/A-G/G	273 (76.7)	50 (70.4)	1		A/A-G/G	177 (49.7)	21 (29.6)	1
	A/G	83 (23.3)	21 (29.6)	1.49 (0.80-2.79)	0.22	A/G	179 (50.3)	50 (70.4)	3.05 (1.66-5.62)	< 0.001
Aditive	---	---	---	1.68 (1.01-2.77)	0.047	---	---	---	1.16 (0.77-1.73)	0.48

¹Odds ratio (OR) adjusted for age, gender and alcohol consumption and smoking habits;

²P values significant at P ≤ 0.05.

Table 3 Association of MTHFR C677T, MTHFR A1298C, MTR A2756G and MTRR A66G polymorphisms with Liver Cirrhosis, adjusted for gender, age, smoking and alcohol consumption

Model	Genotipe	Control n (%)	Case n (%)	1OR (95%CI)	2P-value	Genotipe	Control n (%)	Case n (%)	1OR (95%CI)	2P-value
	MTHFR C677T					MTHFR A1298C
Codominant	C/C	149 (41.9)	48 (41.4)	1		A/A	205 (57.6)	57 (49.1)	1
	C/T	174 (48.9)	55 (47.4)	0.90 (0.56-1.45)	0.56	A/C	116 (32.6)	43 (37.1)	1.27 (0.78-2.07)	0.21
	T/T	33 (9.3)	13 (11.2)	1.37 (0.64-2.95)		C/C	35 (9.8)	16 (13.8)	1.85 (0.92-3.71)
Dominant	C/C	149 (41.9)	48 (41.4)	1		A/A	205 (57.6)	57 (49.1)	1
	C/T-T/T	207 (58.1)	68 (58.6)	0.97 (0.62-1.52)	0.89	A/C-C/C	151 (42.4)	59 (50.9)	1.40 (0.90-2.18)	0.14
Recessive	C/C-C/T	323 (90.7)	103 (88.8)	1		A/A-A/C	321 (90.2)	100 (86.2)	1
	T/T	33 (9.3)	13 (11.2)	1.45 (0.71-2.99)	0.32	C/C	35 (9.8)	16 (13.8)	1.68 (0.86-3.29)	0.14
Overdominant	C/C-T/T	182 (51.1)	61 (52.6)	1		A/A-C/C	240 (67.4)	73 (62.9)	1
	C/T	174 (48.9)	55 (47.4)	0.85 (0.54-1.33)	0.47	A/C	116 (32.6)	43 (37.1)	1.14 (0.72-1.82)	0.58
Aditive	---	---	---	1.07 (0.75-1.51)	0.72	---	---	---	1.33 (0.97-1.83)	0.079
	MTR A2756G					MTRR A66G
Codominant	A/A	263 (73.9)	79 (68.1)	1		A/A	105 (29.5)	37 (31.9)	1
	A/G	83 (23.3)	32 (27.6)	1.52 (0.91-2.53)	0.13	A/G	179 (50.3)	55 (47.4)	0.94 (0.56-1.56)	0.95
	G/G	10 (2.8)	05 (4.3)	2.47 (0.75-8.12)		G/G	72 (20.2)	24 (20.7)	0.91 (0.49-1.71)
Dominant	A/A	263 (73.9)	79 (68.1)	1		A/A	105 (29.5)	37 (31.9)	1
	A/G-G/G	93 (26.1)	37 (31.9)	1.60 (0.98-2.61)	0.06	A/G-G/G	251 (70.5)	79 (68.1)	0.93 (0.58-1.50)	0.77
Recessive	A/A-A/G	346 (97.2)	111 (95.7)	1		A/A-A/G	284 (79.8)	92 (79.3)	1
	G/G	10 (2.8)	5 (4.3)	2.19 (0.68-7.09)	0.21	G/G	72 (20.2)	24 (20.7)	0.95 (0.55-1.64)	0.86
Overdominant	A/A-G/G	273 (76.7)	84 (72.4)	1		A/A-G/G	177 (49.7)	61 (52.6)	1
	A/G	83 (23.3)	32 (27.6)	1.46 (0.88-2.41)	0.15	A/G	179 (50.3)	55 (47.4)	0.97 (0.62-1.52)	0.9
Aditive	---	---	---	1.54 (1.02-2.33)	0.042	---	---	---	0.95 (0.70-1.30)	0.77

¹Odds ratio (OR) adjusted for age, gender and alcohol consumption and smoking habits;

²P values significant at P ≤ 0.05.

Table 4 Interaction between MTHFR C677T, MTHFR A1298C, MTR A2756G and MTRR A66G polymorphisms and smoking habits or alcohol drinking on the risk of hepatocellular carcinoma

	Smoking habits							Alcoholic habit
	No smoker			Smoker			2Pinteraction	Non-alcoholic			Alcoholic			2Pinteraction
	Case	Control	1OR (95%CI)	Case	Control	1OR (95%CI)		Case	Control	1OR(95%CI)	Case	Control	1OR (95%CI)
MTRA2756G
A/A	22	142	1.00	24	121	1.00	0.81	17	137	1.00	29	126	1.00	0.43
A/G-G/G	10	57	1.59 (0.65-3.87)	15	36	1.85 (0.81-4.21)		11	53	2.29 (0.91-5.72)	14	40	1.40 (0.63-3.12)
MTRRA66G
A/A	4	55	1.00	8	50	1.00	0.7	3	53	1.00	9	52	1.00	0.34
A/G-G/G	28	144	3.04 (0.95-9.71)	31	107	2.27 (0.90-5.72)		25	137	4.16 (1.11-15.55)	34	114	1.98 (0.83-4.75)
MTHFRC677T
C/C	15	85	1.00	13	64	1.00	0.55	14	85	1.00	14	64	1.00	0.75
C/T-T/T	17	114	0.80 (0.35-1.81)	26	93	1.13 (0.51-2.52)		14	105	0.86 (0.36-2.06)	29	102	1.04 (0.48-2.22)
MTHFRA1298C
A/A	13	119	1.00	19	85	1.00	0.61	11	113	1.00	21	91	1.00	0.56
A/C-C/C	19	80	2.00 (0.88-4.56)	20	72	1.49 (0.69-3.20)		17	77	2.10 (0.86-5.10)	22	75	1.49 (0.72-3.07)

¹Odds ratio (OR) adjusted for age, gender and alcohol consumption and smoking habits;

²P values significant at P ≤ 0.05.

Table 5 Interaction between MTHFR C677T, MTHFR A1298C, MTR A2756G and MTRR A66G polymorphisms and smoking habits or alcohol drinking on the risk of liver cirrhosis

	Smoking habits							Alcoholic habit
	No smoker			Smoker			2Pinteraction	Non-alcoholic			Alcoholic			2Pinteraction
	Case	Control	1OR(95%CI)	Case	Control	OR+(95%CI)		Case	Control	1OR(95%CI)	Case	Control	1OR (95%CI)
MTRA2756G
A/A	50	142	1.00	29	121	1.00	0.39	36	137	1.00	43	126	1.00	0.73
A/G-G/G	21	57	1.35 (0.72-2.53)	16	36	2.07 (0.97-4.41)		17	53	1.46 (0.72-2.97)	20	40	1.74 (0.89-3.38)
MTRRA66G
A/A	26	55	1.00	11	50	1.00	0.051	14	53	1.00	23	52	1.00	0.42
A/G-G/G	45	144	0.63 (0.35-1.17)	34	107	1.71 (0.77-3.82)		39	137	1.17 (0.56-2.45)	40	114	0.78 (0.42-1.48)
MTHFRC677T
C/C	31	85	1.00	17	64	1.00	0.96	24	85	1.00	24	64	1.00	0.76
C/T-T/T	40	114	0.96 (0.54-1.71)	28	93	0.98 (0.48-2.02)		29	105	1.05 (0.54-2.02)	39	102	0.91 (0.49-1.69)
MTHFRA1298C
A/A	33	120	1.00	24	85	1.00	0.32	23	113	1.00	34	92	1.00	0.56
A/C-C/C	38	79	1.68 (0.95-3.00)	21	72	1.06 (0.53-2.13)		30	77	1.83 (0.95-3.54)	29	74	1.11 (0.61-2.03)

¹Odds ratio (OR) adjusted for age, gender and alcohol consumption and smoking habits;

²P values significant at P ≤ 0.05.

Table 6 Regression analysis of data from multiple logistic analyzed clinical parameters and polymorphisms in patients with hepatocellular carcinoma tumors divided into stages 0 and tumore in stages A and B, C and D according barcelona clinic liver cancer criteria

Variables	Stage 0 e A	Estage B, C e D	OR (95%CI)1	P
	Pacientes n (%)	Pacientes n (%)
Alpha fetoprotein
> 500 ng/mL	22 (84.6)	21 (46.7)	Reference	Reference
< 500 ng/mL	4 (15.4)	24 (53.3)	2.66 (0.55-12.72)	0.22
Hepatitis B virus
Absence	25 (96.2)	38 (84.4)	Reference	Reference
Presence	1 (3.85)	7 (15.6)	4.06 (0.34-48.29)	0.27
Hepatitis C virus
Absence	12 (46.2)	22 (48.9)	Reference	Reference
Presence	14 (53.8)	23 (41.1)	1.43 (0.35-5.78)	0.61
Steatohepatitis
Absence	26 (100)	42 (93.3)	Reference	Reference
Presence	00 (00)	3 (6.7)	3	0.99
Diabetes
Absence	18 (69.2)	32 (71.1)	Reference	Reference
Presence	8 (30.8)	13 (28.9)	0.25 (0.03-1.63)	0.15
Death
No	24 (92.3)	17 (37.8)	Reference	Reference
Yes	2 (7.7)	28 (62.2)	25.3 (3.67-174.38)	0.0012
MTHFR A1298C
AA	10 (38.5)	22 (48.9)	Reference	Reference
AC/CC	16 (61.5)	23 (51.1)	0.93 (0.22-3.86)	0.92
MTHFR C677T
CC	11 (42.3)	17 (37.8)	Reference	Reference
CT/TT	15 (57.7)	28 (62.2)	1.65 (0.34-7.97)	0.53
MTR A2756G
AA	18 (69.2)	28 (62.2)	Reference	Reference
AG/GG	8 (30.8)	17 (37.8)	0.78 (0.18-3.43)	0.75
MTRRA66G
AA	4 (15.4)	8 (17.8)	Reference	Reference
AG/GG	22 (84.6)	37 (82.2)	1.09 (0.16-7.30)	0.93

¹OR: Odds ratio, CI: Confidence interval;

²Statistically significant at P ≤ 0.05;

³Could not calculate due to numerical proximity.