Angiogenesis of hepatocellular carcinoma: An immunohistochemistry study

Table 1 Clinicopathological parameters of the cases used for quantification of angiogenesis

Variable	Number of cases (n = 50)	Percentage
Gender
Male	36	72
Females	14	28
Age (mean ± SD, yr)	64.66 ± 9.6
Age groups
31-50 yr	2	4
51-60 yr	12	24
61-70 yr	21	42
71-80 yr	15	30
Tumor size (mean ± SD, cm)	40.12 ± 40.58
Tumor aspect
Single nodule	36	72
Multifocality	14	28
Tumor grade
G1	8	16
G2	24	48
G3	17	34
G4	1	2
Tumor stage
T1	23	46
T2	27	54
Cirrhosis
Without	32	64
With	18	36
Mallory bodies
Without	36	72
With	14	28
Hepatitis
Without	26	52
With	24	48
Cholestasis
Without	41	82
With	9	18
Steatosis
Without	33	66
With	17	34

Table 2 Antibodies used for quantification of angiogenesis

Antibody	Clone	Company, town, country	Dilution	External control
COX-2	4H12	Novocastra, NewCastle Upon Tyne, United Kingdom	1:100	Brain-multipolar neurons; renal tubes
VEGF	VG1	LabVision, Fremont, CA, United States	1:50	Renal tubes
CD31	JC70A	DAKO,Glostrup, Denmark	1:40	Renal glomerules
CD105	SN6H	LabVision, Fremont, CA, United States	1:50	Tonsils

VEGF: Vascular endothelial growth factor; COX-2: Cyclooxygenase-2.

Table 3 Assessment of COX-2 and VEGF-A expression in HCC

Parameter		COX-2 intensity			P value	VEGF-A intensity			P value
	1+		2+	3+		1+	2+	3+
n (%)	14 (28)		18 (36)	18 (36)		12 (24)	28 (56)	10 (20)
Gender
Males	10		12	14	0.75	8	22	6	0.47
Females	4		6	4		4	6	4
Age groups (yr)
31-50	0		0	2	0.47	0	1	1	0.33
51-60	4		4	4		5	4	3
61-70	5		10	6		4	12	5
71-80	5		4	6		3	11	1
Tumor grade
G1+G2	13		7	12	0.006	6	19	7	0.50
G3+G4	1		11	6		6	9	3
Tumor stage
pT1	7		6	10	0.38	3	12	8	0.03
pT2	7		12	8		9	16	2
Associated lesions
With cirrhosis	8		3	7	0.05	5	9	4	0.81
Without cirrhosis	6		15	11		7	19	6
With Mallory bodies	7		5	2	0.05	4	7	3	0.85
Without Mallory bodies	7		13	16		8	21	7
With hepatitis	7		9	8	0.93	6	14	4	0.85
Without hepatitis	7		9	10		6	14	6

VEGF: Vascular endothelial growth factor; COX-2: Cyclooxygenase-2.

Table 4 Endothelial area assessment, using CD31 and CD105 antibodies in hepatocellular carcinoma

Parameter	Number of cases	CD31-EA(median and range values)	P value		P value
				CD105-EA(median and range values)
Gender
Males	36	7.35 ± 2.16 (2.57-11.18)	0.31	8.91 ± 2.64 (4.06-15.53)	0.44
Females	14	8.06 ± 2.30 (4.17-11.85)		9.59 ± 3.28 (4.23-16.56)
Tumor grade
G1+G2	32	7.69 ± 2.22 (2.57-11.69)	0.53	9.28 ± 2.64 (4.06-16.56)	0.55
G3+G4	18	7.29 ± 2.21 (3.34-11.85)		8.78 ± 3.17 (5.77-15.53)
Tumor stage
pT1	23	7.85 ± 2.33 (2.57-11.69)	0.59	9.47 ± 3.15 (4.06-16.56)	0.39
pT2	27	7.71 ± 2.11 (3.34-11.85)		8.79 ± 2.51 (5.47-14.91)
Associated lesions
With cirrhosis	18	8.29 ± 1.68 (5.64-11.85)	0.07	9.12 ± 2.51 (4.23-14.33)	0.97
Without cirrhosis	32	7.13 ± 2.37 (2.57-11.69)		9.09 ± 3.01 (4.06-16.56)
With Mallory bodies	14	8.01 ± 1.39 (5.56-10.53)	0.36	9.86 ± 2.89 (5.85-15.53)	0.23
Without Mallory bodies	36	7.37 ± 2.44 (2.57-11.85)		8.81 ± 2.77 (4.06-16.56)
With hepatitis	24	7.28 ± 1.99 (2.57-10.53)	0.40	8.26 ± 2.25 (4.23-12.28)	0.04
Without hepatitis	26	7.80 ± 2.39 (3.07-11.85)		9.88 ± 3.09 (4.06-16.56)

EA: Endothelial area.

Table 5 Correlation of the angiogenic immunophenotype, quantified with VEGF-A and COX2, with endothelial area values

	COX-2 intensity			P value	VEGF-A intensity			P value
	1+	2+	3+		1+	2+	3+
EA-CD31	8.66 ± 1.41	7.47 ± 2.29	6.76 ± 2.34	0.04	7.23 ± 1.52	7.53 ± 2.44	8.01 ± 2.32	0.72
EA-CD105	9.22 ± 1.83	9.58 ± 3.56	8.53 ± 2.64	0.53	8.45 ± 1.74	8.95 ± 3.02	10.31 ± 3.15	0.28

EA: Endothelial area; VEGF: Vascular endothelial growth factor; COX-2: Cyclooxygenase-2.