Are gap junction gene connexins 26, 32 and 43 of prognostic values in hepatocellular carcinoma? A prospective study

Table 1 Demographic, clinical and pathological variables in patients with HCC undergoing curative resections (n = 25)

Variables	Patients (n, %)
Age (mean, yr)	56.4 ± 12.6
Male	18 (72)
Child- Pugh class A
Serum AFP < 20 ng/mL	3 (12)
20-103 ng/mL	10 (40)
> 103 ng/mL	12 (48)
HBsAg (+)	12 (48)
Anti-HCV (+)	10 (40)
Diameter of HCC < 3 cm	2 (8)
3-10 cm	10 (40)
> 10 cm	13 (52)
Cirrhosis	20 (80)
Edmondson-Steiner’s grade I	11 (44)
grade II
grade III	14 (56)
grade IV
Complete capsule	9 (36)
Vascular permeation	14 (56)
Daughter nodules	13 (52)
Multinodular HCC	5 (20)
Tumor necrosis	5 (20)
Tumor hemorrhage	1 (4)

AFP: Serum alpha fetoprotein; HBsAg (+): Positive hepatitis B surface antigen; Anti-HCV(+): Positive hepatitis C virus antibody; Edmondson Steiner grade: Differentiation grade.

Table 2 Nucleotide sequences of the primer sets and specific oligonucleotide probes to each type of connexin 5'-noncoding mRNA

Type of connexin mRNA	Primer probes	Nucleotide sequences
Cx 26	Sense	5’ CCGAAGTTCATGAAGGGAGAGAT
	Antisense	5’ GGTCTTTTGGACTTCCCTGAGCA
Cx 32	Sense	5’ CTGCTCTACCCGGGCTATGC
	Antisense	5’ CAGGCTGAGCATCGGTCGCTCTT
Cx 43	Sense	5’ TACCATGCGACCAGTGGTGCGCT
	Antisense	5’GAATTCTGGTTATCATCGTCGGGGAA

Table 3 Correlation of Cx 26, Cx 32, and Cx 43 mRNAs in cancerous and noncancerous liver tissues of 25 patients with HCC and liver tissues of 15 control patients

Tissues	mRNA (mean value)
	Connexin 26	Connexin 32	Connexin 43
HCC patients
Cancerous liver1	0.745	0.775	0.241
Controls2	1.205	1.225	0.100

P < 0.01 for connexin 26 mRNA; P < 0.01 for connexin 32 mRNA; P > 0.05 for connexin 43 mRNA.

Table 4 Predictors of HCC recurrence

Variable	P values
	UV	MV
Sex	1.000	-
Age (yr)	0.030	0.071
Tumor diameter	0.250	-
(< 3 cm, > 10 cm)
Liver cirrhosis	0.009	0.067
Child-Pugh class	0.528	-
Serum AFP	0.744	-
HBsAg (+)	1.000	-
Anti-HCV (+)	1.000	-
Edmondson Steiner grade	< 0.001	0.033
Capsule	< 0.001	0.050
Vascular permeation	< 0.001	0.046
Daughter nodules	< 0.001	0.046
Tumor necrosis	0.046	-
Tumor hemorrhage	0.046	-
Connexin 26 mRNA	< 0.001	0.033
Connexin 32 mRNA	< 0.001	0.033
Connexin 43 mRNA	0.280	0.071

UV: Univariate analysis; MV: Multivariate analysis; AFP: Serum alpha fetoprotein; HbsAg (+): Positive hepatitis B sur-face antigen; Anti-HCV (+): Positive hepatitis C virus antibody; Edmondson Steiner grade: Differentiation grades I, II vs III, IV.

Table 5 Correlation between clinical and pathological vari-ables and recurrence-related mortality

Parameters	P values
	UV	MV
Sex	0.888	-
Age (yr)	0.005	0.356
Tumor diameter	0.324	-
(< 3 cm, > 10 cm)
Liver cirrhosis	0.030	0.324
Child-Pugh class	0.548
HBsAg (+)	0.956	-
Anti-HCV (+)	0.785	-
Edmondson Steiner grade	0.000	0.031
Capsule	0.000	0.053
Vascular permeation	0.000	0.048
Daughter nodules	0.039	0.048
Multinodular HCC	0.007	0.324
Tumor necrosis	0.373	-
Tumor hemorrhage	0.356	-
Connexin 26 mRNA	< 0.001	0.031
Connexin 32 mRNA	< 0.001	0.031
Connexin 43 mRNA	0.461	-

UV: Univariate analysis; MV: Multivariate analysis; AFP: Se-rum alpha fetoprotein; HBsAg (+): Positive hepatitis B surface antigen; Anti-HCV(+): Positive hepatitis C virus antibody; Edmondson Steiner grade: Differentiation grades I, II vs III, IV; n.s: Not significant.