Evaluation of advanced protein oxidation and RAGE gene variants in the risk of laryngeal cancer

Figures & data

Table 1. Clinical characteristics of patients with laryngeal carcinoma.

Tumour localization (n, %)	Glottic	81 (67.5 %)
	Supraglottic	39 (32.5 %)
Differentiation (n, %)	Well	20 (16.7 %)
	Moderate	80 (66.6 %)
	Poor	20 (16.7 %)
Tumour grade (n, %)	T1	11 (9.1 %)
	T2	21 (17.5 %)
	T3	68 (56.8 %)
	T4	20 (17.6 %)
Lymphatic invasion (n, %)	Present (n1 + n2 + n3)	53 (44.2 %)
	Absent (n0)	67 (55.8 %)
Perineural invasion (n, %)	Present	41 (34.2 %)
	Absent	79 (65.8 %)
Muscle invasion (n, %)	Present	87 (72.8 %)
	Absent	33 (27.2 %)

Table 2. Distribution of genotype and allele frequencies of RAGE −374 T/A (rs1800624) and RAGE Gly82Ser (G > A; rs2070600) polymorphisms in the study groups.

(n, %)	Control group (n = 120)	Patient group (n = 120)
RAGE rs1800624 (−374 T > A)
Genotypes
TT	32 (26.7%)	36 (30.0%)
TA	48 (40.0%)	56 (46.7%)
AA	40 (33.3%)	28 (23.3%)
Alleles
T allele	112 (46.7 %)	128 (53.3 %)
A allele	128 (53.3 %)	112 (46.7 %)
RAGE rs2070600 (Gly82Ser; G > A)
Genotypes
GlyGly (GG)	115 (95.8%)	78 (65.0%)
GlySer (GA)	5 (4.2%)	42 (35.0%)
SerSer (AA)	–	–
Alleles
Gly (G) Allele	235 (98.0%)	198 (82.5%)
Ser (A) Allele	5 (2.0%)	42 (17.5%)*

The difference between the groups was analysed by χ²) test.

*Statistical significance (p < 0.05).

n, number of individuals.

Table 3. Oxidant and antioxidant levels in the study groups and their correlations in LC patients group.

Oxidant and antioxidant parameters	Control group	Patient group	p value	Correlations in LC patients group
				PCO	AOPP	LPH	T-SH	NP-SH	P-SH
PCO (nmol/mg)	0.9 ± 0.05	1.2 ± 0.1	< 0.001	1
AOPP (μmol/L chloramine-T equivalent)	40.2 ± 3.5	56.7 ± 3.7	0.002	0.063	1
LPH (μmol/mg pr)	1.2 ± 0.1	3.9 ± 0.3	< 0.001	0	0.236	1
T-SH (nmol/mg pr)	17.1 ± 1.1	15.2 ± 0.8	0.166	0.227	0.161	0.359**	1
NP-SH (nmol/mg pr)	3.5 ± 0.1	3.3 ± 0.2	0.619	0.004	−0.163	−0.122	0.068	1
P-SH (nmol/mg pr)	13.7 ± 1.1	11.8 ± 0.8	0.17	0.219	0.185	0.379**	0.966**	−0.192	1
Cu, Zn-SOD (U/mg pr)	6.1 ± 0.2	7.2 ± 0.2	0.001	0.04	−0.1	−0.390**	−0.164	0.029	−0.167

The difference between the groups was analysed by Student’s t test. p < 0.05 indicates statistical significance. **Significance of correlation at the 0.01 level. PCO, protein carbonyl groups; AOPP, advanced oxidation protein products; LPH, lipid hydroperoxides; T-SH, total thiol; NP-SH, non-protein thiol; P-SH, protein thiol; Cu,Zn-SOD, copper, zinc superoxide dismutase; pr, protein.

Figure 1. ROC Curve analysis of significant oxidant and antioxidant parameters.

MedCalc Statistical Software (MedCalc Software Ltd, Belgium/WEB address: [email protected]) program was used to obtain ROC curve analysis. AUC, area under curve; PCO, protein carbonyl; AOPP, advanced oxidation protein products; T-SH, total thiol; NP-SH, non-protein thiol; P-SH, protein thiol; LPH, lipid hydroperoxides; Cu, Zn-SOD, copper, zinc superoxide dismutase. P shows statistical significance as p < 0.05.

Table 4. Logistic regression analysis of risk factors that may affect the pathogenesis of laryngeal cancer between the patient and control groups.

A. According to the levels of significant oxidant and antioxidant parameters
Dependent variable	Independent variables	Exp (B) (OR)	p value	95% CI
LC patients/controls	Model 1
	LPH	2.6	< 0.001	1.6–4.2
	PCO	3.3	0.129	0.7–14.9
	SOD	2.0	0.003	1.3–3.1
	AOPP	1.1	0.011	1.0–1.1
	Model 2
	LPH	2.6	<0.001	1.6–4.2
	SOD	2.1	0.001	1.4–3.2
	AOPP	1.1	0.006	1.0–1.1
	Model 3
	PCO	4.0	0.035	1.1–14.1
	SOD	1.5	0.016	1.1-2.2
	AOPP	1.1	0.001	1.0–1.1

Table

B. According to cut-off values of the independent variables
Dependent variable	Independent variables	Exp (B) (OR)	p value	95% CI
LC patients/controls	LPH (≥1.73 μmol/mg pr →1)	4.8	0.004	1.6–14.2
	PCO (≥0.955 nmol/mg pr →1)	3.1	0.040	1.1–9.0
	SOD (≥6.495 U/mg pr →1)	8.3	0.001	2.3–29.6
	AOPP (≥44.8 μmol/L chloramine-T equivalent →1)	7.5	0.002	2.1–26.7

Binary logistic regression analysis was performed according to the levels of significant oxidant and antioxidant parameters as the independent variables in A, and according to cut-off values of the independent variables in B in which the risk status was above the cut-off value and each of these parameters were coded as 1. OR, odds ratio; CI, Confidence interval. p < 0.05 denotes statistical significance.

Table 5. Distribution of oxidative and antioxidative parameters in the prognosis of laryngeal carcinoma and among RAGE rs1800624 and rs2070600 variants.

RAGE	PCO (nmol/mg pr)	AOPP (μmol/L chloramine-T equivalent)	LPH (μmol/mg pr)	T-SH (nmol/mg pr)	NP-SH (nmol/mg pr)	P-SH (nmol/mg pr)	Cu,Zn-SOD (U/mg pr)
Early stage (T1 +T2)	1.1 ± 0.1	54.9 ± 5.1	3.8 ± 0.6	10.3 ± 1.0	2.4 ± 0.3	8.0 ± 0.9	7.5 ± 0.3
Advanced stage (T2 + T3)	1.2 ± 0.1	57.4 ± 4.8	3.5 ± 0.3	16.9 ± 0.9	3.8 ± 0.3	13.2 ± 1.0	7.0 ± 0.3
p value	0.343	0.721	0.656	< 0.001	< 0.001	0.001	0.269
Lymphatic invasion (+)	1.2 ± 0.1	60.7 ± 7.2	3.1 ± 0.4	14.9 ± 1.0	3.8 ± 0.3	11.1 ± 1.1	7.5 ± 0.4
(−)	1.2 ± 0.1	53.4 ± 3.5	4.1 ± 0.4	15.4 ± 1.2	3.0 ± 0.3	12.3 ± 1.2	6.9 ± 0.3
p value	0.972	0.359	0.074	0.750	0.035	0.472	0.159
Perineural invasion (+)	1.2 ± 0.1	57.2 ± 7.6	3.3 ± 0.5	16.8 ± 1.5	3.1 ± 0.3	13.8 ± 1.5	7.4 ± 0.5
(−)	1.2 ± 0.1	56.4 ± 4.3	3.7 ± 0.3	14.7 ± 1.0	3.4 ± 0.2	11.2 ± 1.0	7.1 ± 0.2
p value	0.648	0.897	0.530	0.269	0.498	0.188	0.663
Muscle invasion (+)	1.3 ± 0.1	60.3 ± 5.1	4.1 ± 0.4	16.2 ± 1.1	3.4 ± 0.3	12.8 ± 1.1	7.3 ± 0.3
(−)	1.1 ± 0.1	44.6 ± 3.3	2.6 ± 0.4	11.6 ± 1.2	3.1 ± 0.2	8.6 ± 1.1	6.7 ± 0.4
p value	0.298	0.012	0.011	0.004	0.316	0.010	0.011
rs1800624
TT	1.3 ± 0.1	56.1 ± 3.2	4.0 ± 0.5	13.8 ± 1.4	2.9 ± 0.3	11.0 ± 1.5	7.2 ± 0.3
TA	1.2 ± 0.1	52.8 ± 6.5	2.6 ± 0.4	16.8 ± 1.1	4.0 ± 0.3	12.4 ± 1.2	6.9 ± 0.2
AA	1.1 ± 0.2	68.6 ± 10.4	5.2 ± 0.4	14.6 ± 2.4	2.6 ± 0.3	12.0 ± 2.1	7.3 ± 0.6
p value	0.401	0.328	0.001	0.37	0.01	0.78	0.924
T Allele	1.3 ± 0.1	54.3 ± 3.9	3.2 ± 0.3	15.3±.0.9	3.5 ± 0.2	11.8 ± 0.9	7.1 ± 0.2
p value (T allele vs. AA )	0.062	0.149	0.001	0.748	0.086	0.940	0.789
A Allele	1.2 ± 0.1	57.0 ± 5.6	3.4 ± 0.3	15.1 ± 1.0	3.6 ± 0.3	12.3 ± 1.0	7.1 ± 0.3
p value (A allele vs. TT)	0.598	0.892	0.343	0.234	0.105	0.491	0.844
rs2070600
GlyGly (GG)	1.3 ± 0.1	57.1 ± 4.0	3.8 ± 0.3	16.0 ± 1.0	3.5 ± 0.3	12.4 ± 1.0	6.9 ± 0.2
GlySer (GA)	1.2 ± 0.1	55.7 ± 8.2	3.3 ± 0.5	13.3 ± 1.4	2.9 ± 0.3	10.5 ± 1.4	7.8 ± 0.4
SerSer (AA)	–	–	–	–	–	–	–
p value	0.67	0.869	0.408	0.128	0.175	0.28	0.051
Gly (G) Allele	1.2 ± 0.1	56.7 ± 3.7	3.6 ± 0.3	15.2±.0.8	3.3 ± 0.2	11.8 ± 0.8	7.2 ± 0.2
p value (G allele vs. AA)]	N/A	N/A	N/A	N/A	N/A	N/A	N/A
Ser (A) Allele	1.2 ± 0.1	55.7 ± 8.2	3.3 ± 0.5	13.3 ± 1.4	2.9 ± 0.3	10.5 ± 1.4	7.8 ± 0.4
p value (A allele vs. GG)	0.670	0.869	0.408	0.128	0.175	0.280	0.051

The comparison of the levels of oxidative and antioxidative parameters among the clinical outcome were analysed with Student’s t test. The genotypic and allelic differences of oxidative stress parameters were examined by one-way ANOVA and Student’s t test, respectively. Boldface p values denote statistical significance of p < 0.05. PCO, protein carbonyl; AOPP, advanced oxidation protein product; T-SH, total thiol; NP-SH, non-protein thiol; P-SH, protein thiol; LPH, lipid hydroperoxides; Cu, Zn-SOD, copper, zinc superoxide dismutase; pr, protein; vs, versus; N/A, not available.