Expression of angiopoietins, Tie2 and vascular endothelial growth factor in angiogenesis and progression of hepatocellular carcinoma

Table 1 Demographic data and clinicopathological features in 38 cases of HCC

No.	Sex	Age	Tumor size(cm)	Tumorcapsule	Metastasis	Portal vein invasion	Histologicalgrade	Liver cirrhosis
1	M	51	< 5	+	-	-	G2	-
2	M	49	< 5	+	-	-	G3	+
3	M	31	> 10	-	-	-	G2	+
4	M	59	5-10	+	-	-	G3	+
5	F	55	> 10	-	-	-	G1	+
6	M	65	> 10	-	-	+	G3	+
7	M	56	> 10	-	+	+	G2	+
8	M	51	> 10	-	-	+	G3	+
9	F	49	< 5	+	-	-	G1	-
10	M	50	5-10	-	-	-	G2	+
11	M	53	> 10	-	-	+	G2	+
12	M	43	< 5	+	-	-	G3	+
13	M	36	5-10	+	-	-	G2	+
14	F	46	5-10	-	-	-	G1	+
15	F	45	5-10	-	-	-	G1	+
16	M	52	5-10	-	-	-	G2	+
17	F	50	5-10	-	-	-	G1	+
18	F	52	< 5	+	-	-	G1	+
19	F	61	> 10	-	+	+	G1	+
20	M	36	5-10	-	-	-	G2	+
21	M	46	< 5	+	-	-	G2	+
22	M	48	> 10	-	-	+	G2	+
23	F	63	< 5	+	-	-	G1	+
24	M	35	5-10	+	-	-	G2	-
25	M	74	> 10	-	+	+	G3	+
26	F	55	5-10	-	-	-	G1	+
27	M	59	5-10	-	-	-	G2	+
28	M	30	< 5	+	-	-	G3	+
29	F	56	5-10	-	-	-	G1	+
30	M	60	5-10	-	-	-	G2	+
31	F	65	< 5	+	-	-	G1	+
32	M	59	> 10	-	+	+	G2	+
33	M	61	> 10	-	+	+	G2	+
34	M	55	< 5	-	-	-	G2	-
35	M	48	5-10	+	-	-	G3	+
36	M	49	> 10	-	-	+	G3	+
37	F	50	5-10	-	-	-	G1	+
38	M	58	> 10	-	+	+	G3	+

Table 2 Primer sequences of each gene for real-time RT-PCR assay

Target gene	Sense	Antisense
Ang-1	ACTGT GCAGA TGTAT ATCAA GC	GTGGA ATCTG TCATA CTGTG AA
Ang-2	GGAAG ACAAG CACAT CATCC	AGTAA GCCTG ATTCC CTTCC
Tie2	TCTGT GCTGT TCCTT CTTGC	CTTGA GTAAC TTCCA GCGGA
VEGF	AGCTA CTGCC ATCCA ATCGC	GGGCG AATCC AATTC CAAGA G
G3PDH	GTCAA CGGAT TTGGT CTGTA TT	AGTCT TCTGG GTGGC AGTGA T

Table 3 Relation between gene expression pattern and clinicopathological parameters (mean ± SD)

Clinical parameters	Ang-2	Ang-1	Ang-2/1	Tie2	VEGF
Tumor size (cm)
< 5 (n = 10)	0.267 ± 0.113	0.148 ± 0.089	2.175 ± 1.025	1.686 ± 0.550	0.552 ± 0.205
≥ 5 (n = 28)	0.748 ± 0.422	0.212 ± 0.082	3.880 ± 2.067	1.570 ± 0.990	1.213 ± 0.537
Histlogical grade
G1 (n = 12)	0.509 ± 0.320	0.168 ± 0.064	3.106 ± 1.409	1.511 ± 0.574	0.916 ± 0.497
G2 (n = 16)	0.633 ± 0.395	0.194 ± 0.081	3.543 ± 1.925	1.470 ± 0.726	1.095 ± 0.604
G3 (n = 10)	0.737 ± 0.512	0.234 ± 0.101	3.645 ± 2.470	1.921 ± 0.138	1.097 ± 0.585
Portal vein invasion
positive (n = 11)	1.152 ± 0.376	0.232 ± 0.053	5.096 ± 1.691	1.880 ± 1.494	1.791 ± 0.423
negative (n = 27)	0.405 ± 0.163	0.180 ± 0.092	2.753 ± 1.631	1.488 ± 0.500	0.732 ± 0.254
Metastasis
positive (n = 6)	1.320 ± 0.366	0.235 ± 0.061	5.912 ± 1.847	2.262 ± 1.734	2.007 ± 0.214
negative (n = 32)	0.490 ± 0.271	0.187 ± 0.088	2.967 ± 1.600	1.148 ± 0.619	0.857 ± 0.415
Tumor capsule
positive (n = 13)	0.308 ± 0.135	0.178 ± 0.112	2.078 ± 0.887	1.733 ± 0.424	0.579 ± 0.198
negative (n = 25)	0.783 ± 0.429	0.203 ± 0.068	4.135 ± 1.998	1.529 ± 1.056	1.278 ± 0.558