Differential expression of cell cycle regulators in HCV-infection and related hepatocellular carcinoma

Table 1 Clinical and laboratory data of all studied cases (mean ± SD) n(%)

Parameters	Control	CHC without cirrhosis	CHC with cirrhosis	HCV-related HCC
	(n = 15)	(n = 50)	(n = 30)	(n = 30)
Age	45.0 ± 7.5	47.4 ± 9.3	51.3 ± 5.9	48.9 ± 7.2
Male/female ratio	2/1	7/3	2/1	2/1
Pallor	0 (0)	2 (4.0)	5 (16.5)	8 (26.6)
Jaundice	0 (0)	3 (6.0)	6 (20.0)	13 (43.3)
Palmer erythema	0 (0)	0 (0)	15 (50.0)	17 (56.6)
Spider naevi	0 (0)	0 (0)	13 (43.3)	16 (53.3)
Lower limb edema	0 (0)	0 (0)	16 (53.3)	10(33.3)
Child classification
A	0 (0)	50 (100)	8 (26.6)	1 (3.3)
B		0 (0)	9 (30.0)	11 (36.6)
C		0 (0)	13 (43.3)	18 (60.0)
ALT (IU/L)	32.6 ± 4.2	63.2 ± 29.7	51.8 ± 4.3	78.1 ± 16.3
AST (IU/L)	31.1 ± 5.1	49.1 ± 18.3	46.4 ± 5.1	68.3 ± 12.4
Albumin (g/dL)	4.4 ± 0.5	3.80 ± 0.3	2.90 ± 0.7	2.80 ± 0.4
PT conc.	97.6 ± 3.4	89.7 ± 5.8	53.4 ± 11.2	68.5 ± 3.8

Table 2 Immunohistochemical reactivity for Cyclin D1 in all studied cases (mean ± SE) n(%)

Groups n = 125	Staining pattern	Positive expression	Immunohistochemical reactivity			Hepatic expression
			Mild (< 5%)	Moderate (5%-30%)	Marked (> 30%)
Control (n = 15)	Nuclear	3 (20.0)	3 (100)	0 (0)	0 (0)	1.33 ± 0.33
CHC without cirrhosis (n = 50)	Nuclear	33 (66.0)	25 (75.8)	8 (24.2)	0 (0)	6.60 ± 0.72a
CHC with cirrhosis (n = 30)	Nuclear	21 (70.0)	15 (71.4)	6 (28.6)	0 (0)	9.95 ± 1.70ab
HCV-related HCC (n = 30)	Nuclear	30 (100)	12 (40.0)	12 (40.0)	6 (20.0)	19.85 ± 3.90abc
Well-differentiated HCC (n = 15)	Nuclear	15 (100)	9 (60.0)	6 (40.0)	0 (0)	10.10 ± 2.20ab
Poorly-differentiated HCC (n = 15)	Nuclear	15 (100)	3 (20.0)	6 (40.0)	6 (40)	31.60 ± 6.20abcd

^aP < 0.001 vs controls;

^bP < 0.01 vs CHC without cirrhosis;

^cP < 0.001 vs LC;

^dP < 0.001 vs well differentiated HCC.

Table 3 Immunohistochemical reactivity for cyclin E in patients with well-differentiated and poorly-differentiated (HCC) (mean ± SE) n(%)

Groups	Positive expression	Immunohistochemical reactivity			Stained cells
		Mild < 5%	Moderate 5%-49%	Marked 50%-100%
Well-differentiated HCC (n = 15)	6 (40.0)	2 (33.3)	4 (66.7)	0 (0)	25.3 ± 5.9
Poorly-differentiated HCC (n = 15)	6 (40.0)	0 (0)	0 (0)	6 (100)	82.5 ± 2.1a

^aP < 0.01 vs well differentiated HCC.

Table 4 Immunohistochemical reactivity for p21 in all studied cases (mean ± SE)

Groups	Staining pattern	Hepatic expression
Control (n = 15)	Nuclear	1.5 ± 0.5
CHC without cirrhosis (n = 50)	Nuclear	6.5 ± 0.2a
CHC with cirrhosis (n = 30)	Nuclear	7.2 ± 0.8a
HCV-related HCC (n = 30)	Nuclear	4.9 ± 0.1ab
Well-differentiated HCC (n = 15)	Nuclear	5.6 ± 0.16
Poorly-differentiated HCC (n = 15)	Nuclear	4.2 ± 0.13

^aP < 0.01 vs controls;

^bP < 0.05 vs LC.

Table 5 Immunohistochemical reactivity for p27 in all studied cases (mean ± SE) n(%)

Groups n = 125	Staining pattern	Immunohistochemical reactivity		Hepatic expression
		Weak (< 50%)	Marked (> 50%)
Control (n = 15)	Nuclear/mixed	9 (60.0)	6 (40.0)	44.4 ± 1.9
CHC without cirrhosis (n = 50)	Nuclear/cytoplasmic	5 (10.0)	45 (90.0)	59.8 ± 1.9a
CHC with cirrhosis (n = 30)	Nuclear/mixed	0 (0)	30 (100)	65.9 ± 1.6a
HCV-related HCC (n = 30)	Nuclear/mixed	21 (70.0)	9 (30.0)	34.1 ± 2.7abc
Well-differentiated HCC (n = 15)	Nuclear	7 (46.7)	8 (53.3)	36.2 ± 5.2abc
Poorly-differentiated HCC (n = 15)	Mixed	15 (100)	0 (0)	32.0 ± 1.7abc

^aP < 0.01 vs controls;

^bP < 0.001 vs CHC without cirrhosis;

^cP < 0.001 vs LC.

Table 6 Immunohistochemical reactivity for Rb/p105 in all studied cases (mean ± SE) n(%)

Groups n = 125	Staining pattern	Positive expression	Immunohistochemical reactivity			Hepatic expression
			Mild	Moderate	Marked
			(1%-30%)	(31%-60%)	(61%-100%)
Control (n = 15)	Negative	0 (0)	0 (0)	0 (0)	0 (0)	0.00 ± 0.00
CHC without cirrhosis (n = 50)	Nuclear	30 (60.0)	27 (90.0)	3 (10.0)	0 (0)	28.7 ± .1
CHC with cirrhosis (n = 30)	Nuclear	15 (50.0)	5 (33.3)	10 (66.7)	0 (0)	40.9 ± 2.8a
HCV-related HCC (n = 30)	Nuclear	21 (70.0)	0 (0)	12 (57.1)	9 (42.9)	61 ± 3.7ab
Well-differentiated HCC (n = 15)	Nuclear	12 (80.0)	0 (0)	12 (100)	0 (0)	67.3 ± 4.8ab
Poorly-differentiated HCC (n = 15)	Nuclear	9 (60.0)	0 (0)	0 (0)	9 (100)	55.7 ± 1.4abc

^aP < 0.01 vs CHC without cirrhosis;

^bP < 0.001 vs LC;

^cP < 0.01 vs well-differentiated carcinoma.

Table 7 Correlation analysis of different parameters in HCCs

Parameter	HCV-related HCC
	r	P
Cyclin D1 # Cyclin E	0.61	< 0.01
Cyclin D1 # p21	-0.51	< 0.001
Cyclin D1 # p27	-0.65	< 0.001
Cyclin E # Rb/p105	0.62	< 0.01
Cyclin E # p21	-0.64	< 0.01
p21 # p27	0.47	< 0.001
p21 # Rb/p105	-0.42	< 0.05