Circulating microRNA 9-3p and serum endocan as potential biomarkers for hepatitis C virus-related hepatocellular carcinoma

Table 1 Demographic and laboratory data of the study participants

Variables		Group I	Group II	Group III	P value
		HCC	Chronic liver disease	Control
		n = 35	n = 33	n = 32
Gender
Male, n (%)		30 (85.7)	23 (69.7)	27 (84.4)	NS
Female, n (%)		5 (14.3)	10 (30.3)	5 (15.6)
Age (yr)
mean ± SD		55.2 ± 5.2	52.7 ± 5.3	52.8 ± 5.6	NS
ALT (IU/L), median (IQR)		50.0 (35.0-55.0)	34.0 (28.0-50.0)	29.7 (23.5-31.7)	P < 0.001	aP = 0.005; bP < 0.001; cP = 0.003
AST (IU/L), median (IQR)		52.0 (39.0-70.0)	42.0 (32.0-57.0)	32.8 (30.0-36.0)	< 0.001	aP = 0.060; bP < 0.001; cP < 0.001
Hb (mg/dL), mean ± SD		13.1 ± 1.7	12.5 ± 1.6	13.5 ± 1.0	0.025	aP = 0.260; bP = 0.438; cP = 0.019
Platelets, (× 10³/μL), median (IQR)		141.0 (104.5-193.5)	162.0 (134.0-213.0)	197.5 (180.5-246.0)	0.002	aP = 0.225; bP < 0.001; cP = 0.022
Serum ALB (g/dL), mean ± SD		3.6 ± 0.7	3.8 ± 0.6	4.1 ± 0.4	0.001	aP = 0.382; bP = 0.001; cP = 0.050
INR, mean ± SD		1.3 ± 0.2	1.1 ± 0.4	0.8 ± 0.2	< 0.001	aP = 0.002; bP = 0.001; cP = 0.001
α-fetoprotein (ng/mL), median (IQR)		240.0 (28.2-635.0)	124.0 (108.9-166.0)	17.4 (14.0-24.0)	< 0.001	aP = 0.895; bP < 0.001; cP < 0.001
Endocan (pg/mL), median (IQR)		3450.0 (3188.5-4135.0)	1934.0 (1450.0-2257.0)	878.5 (850.0-1188.0)	< 0.001	aP < 0.001; bP < 0.001; cP = 0.001
microRNA 9-3p, median (IQR)		0.03 (0.02-0.05)	0.42 (0.29-1.35)	1.70 (1.40-2.15)	< 0.001	aP < 0.001; bP < 0.001; cP < 0.001

^aP: P value for comparing between HCC and chronic liver disease.

^bP: P value for comparing between HCC and control.

^cP: P value for comparing between chronic liver disease and control. Statistically significant at P ≤ 0.05. SD: Standard deviation; AST: Aspartate aminotransferase; ALT: Alanine aminotransferase; Hb: Hemoglobin; ALB: Albumin; INR: International normalized ratio; HCC: Hepatocellular carcinoma; NS: Not significant; IQR: Interquartile range.

Table 2 Clinical characteristics of tumors in the hepatocellular carcinoma group, n = 35

Number of the focal lesions	n (%)
Single	16 (45.7)
Multiple	19 (54.3)
Tumor size in cm
Small < 3	7 (20.0)
Medium 3-5	15 (42.9)
Large > 5	13 (37.1)
Location of the focal lesions
Rt. Lobe	19 (54.3)
Lt. Lobe	8 (22.9)
Both	7 (20.0)
Caudate lobe	1 (2.9)
BCLC stage
A	4 (11.4)
B	18 (51.4)
C	13 (37.1)
Vascular invasion
Negative	25 (71.4)
Positive	10 (28.6)
LN metastasis
Negative	28 (80.0)
Positive	7 (20.0)
Ascites
No	25 (73.5)
Mild	8 (23.5)
Moderate	1 (2.9)
Child Pugh classA	22 (62.9)
B	13 (37.1)
C	0 (0)
Distant metastasis
No	23 (65.7)
Yes	12 (34.3)

Rt.: Right; Lt.: Left; BCLC: Barcelona Clinic Liver Cancer; LN: Lymph node.

Table 3 Correlations between microRNA 9-3p levels and clinical data in hepatocellular carcinoma group

	n	microRNA 9-3p
		Median (IQR)		P value
Vascular invasion
Negative	25	0.04 (0.02-0.26)		0.002
Positive	10	0.02 (0.02-0.02)
LN metastasis
Negative	28	0.04 (0.02-0.17)		0.072
Positive	7	0.02 (0.02-0.03)
Distant metastasis
No	23	0.04 (0.03-0.26)		< 0.001
Yes	12	0.02 (0.02-0.02)
Child Pugh class
A	22	0.03 (0.02-0.04)		0.389
B	13	0.04 (0.02-0.26)
Tumor number
Single	16	0.03 (0.02-0.17)		0.935
Multiple	19	0.03 (0.02-0.05)
Tumor size in cm
Small < 3	7	0.03 (0.02-0.15)		0.852
Medium 3-5	15	0.03 (0.02-0.06)
Large > 5	13	0.04 (0.02-0.04)
Tumor site
Rt lobe	19	0.04 (0.02-0.06)		0.432
Lt lobe	8	0.04 (0.03-0.15)
Both	7	0.02 (0.02-0.04)
Caudate lobe	1
BCLC stage
A	4	0.26 (0.17-0.26)		< 0.001
B	18	0.04 (0.03-0.05)
C	13	0.02 (0.02-0.02)

Rt.: Right; Lt.: Left; BCLC: Barcelona Clinic Liver Cancer; IQR: Interquartile range; LN: Lymph node.

Table 4 Univariate and multivariate regression analyses for the parameters affecting metastasis in hepatocellular carcinoma group

	Univariate		Multivariate
	P value	OR (95%CI)	P value	OR (95%CI)
microRNA 9-3p	0.008	0.193 (0.057-0.653)	0.041	0.291 (0.089-0.951)
Endocan	0.023	1.002 (1.000-1.003)	0.358	1.001 (0.999-1.002)

Statistically significant at P ≤ 0.05. OR: Odds ratio; CI: Confidence interval.