Expression of heat shock proteins (HSP27, HSP60, HSP70, HSP90, GRP78, GRP94) in hepatitis B virus-related hepatocellular carcinomas and dysplastic nodules

Table 1 Expression of HSPs in hepatocarcinogenesis obtained by IHC analysis.

HSP	Percentage of positive cells (%)	NL (n = 57)	DN		HCC				P (r)
			LGDN (n = 11)	HGDN (n = 8)	E-HCC (n = 5)	GI (n = 12)	GII (n = 11)	GIII (n = 10)
HSP27	0-5	42	11	6		5	2	2
	6-25	12		2	4	5	3	3	0.0001
	26-50	3					2	1	(0.600)
	51-75				1	1	2
	>75					1	2	4
HSP60	0-5							1
	6-25								0.0121
	26-50	3					3		(-0.329)
	51-75	2						1
	>75	52	11	8	5	12	8	8
HSP70	0-5	57	11	8	3	4	3	2
	6-25				1	3	2		0.0001
	26-50				1	3	2	1	(0.669)
	51-75					1	2	4
	>75					1	2	3
HSP90	0-5	56	10	8	5	8	4
	6-25	1	1			2	4	3	0.0001
	26-50					2	2	3	(0.707)
	51-75						1	2
	>75					2
GRP78	0-5	5	6	1	1	1
	6-25	22	5	7	4	5	2		0.0001
	26-50	17				1	2	1	(0.802)
	51-75	9				3	2	1
	>75	4				2	5	8
GRP94	0-5	16	3	3	2	1	2
	6-25	26	7	4	2	2			0.0001
	26-50	9	1	1	1	7	3	2	(0.708)
	51-75	3				1	3
	>75	3				1	3	8

NL: non-tumorous liver; DN: dysplastic nodule; LGDN: low-grade DN; HGDN: high-grade DN; HCC: hepatocellular carcinoma; E: early; G: Edmondson-Steiner’s grade; n: number of cases; P: Spearman correlation (from LGDN to HCC GIII); (r): Correlation coefficient;

¹Value was statistically significant.

Table 2 Expression of HSPs in hepatocarcinogenesis obtained by dot immunoblot analysis.

HSP	Tumor/ non-tumor	DN		HCC				P (r)
		LGDN (n = 11)	HGDN (n = 8)	E-HCC (n = 5)	GI (n = 12)	GII (n = 11)	GIII (n = 10)
HSP27	< 0.5		1	1
	0.5-1.5	9	6		3	4	4	0.0021
	1.5-2.5	2	1	2	6	2	2	(0.407)
	> 2.5			2	3	5	4
HSP60	< 0.5				1
	0.5-1.5	10	6	4	4	5	4	0.0041
	1.5-2.5	1	2	1	4	6	4	(0.372)
	> 2.5				3		2
HSP70	< 0.5
	0.5-1.5	6	7	3	3	2	5	0.100
	1.5-2.5		5	2	4	8	4	(0.220)
	> 2.5		1		5	1	1
HSP90	< 0.5			1
	0.5-1.5	5	6	3	2	2	2	0.0071
	1.5-2.5	6	2	1	4	7	6	(0.353)
	> 2.5				6	2	2
	GRP78 < 0.5		1				1
	0.5-1.5	5	3	3	2	3	2	0.0071
	1.5-2.5	5	4	1	4	3		(0.355)
	> 2.5	1		1	6	5	7
GRP94	< 0.5						1
	0.5-1.5	7	1	2	1	5	4	0.571
	1.5-2.5	2	6	3	5	4	1	(0.077)
	> 2.5	2	1		6	2	4

DN: dysplastic nodule; LGDN: low-grade DN; HGDN: high-grade DN; HCC: hepatocellular carcinoma; E: early; G: Edmondson-Steiner’s grade; n: number of cases; P: Spearman Correlation; (r): Correlation coefficient;

¹Value was statistically significant.

Table 3 Enhancement of HSP expression relative to non-tumorous tissue in hepatocarcinogenesis obtained by dot immunoblot analysis.

Group	n	Enhancement of expression (mean±SD)
		HSP27	HSP60	HSP70	HSP90	GRP78	GRP94
LGDN	11	1.25±0.31	1.07±0.43	1.37±0.45	1.55±0.45	1.65±0.85	1.61±1.02
HGDN	8	1.22±0.72	1.15±0.44	1.32±0.58	1.20±0.32	1.42±0.71	2.45±1.87
E-HCC	5	2.52±1.57	0.93±0.41	1.18±0.49	1.04±0.64	1.72±1.16	1.63±0.54
HCC GI	12	2.17±1.06	1.96±1.20	2.69±1.72	2.76±1.70	7.41±8.65	13.70±25.68
HCC GII	11	3.26±3.19	1.58±0.41	1.80±0.55	1.95±0.56	2.65±1.30	1.87±0.90
HCC GIII	10	2.68±2.34	2.14±1.58	1.66±0.84	2.22±1.33	4.54±4.79	3.02±2.70
P		0.0181	0.0041	0.123	0.0211	0.0051	0.373

LGDN: low-grade dysplastic nodule; HGDN: high-grade dysplastic nodule; E: early; HCC: hepatocellular carcinoma; G: Edmondson-Steiner’s grade; n: number of cases; SD: standard deviation; P: Jonckheere-Terpstra test;

¹Value was statistically significant.

Table 4 Correlation between HSP immunostaining and prognostic factors in 38 HCCs.

	P
	HSP27	HSP60	HSP70	HSP90	GRP78	GRP94
Histological grade
E–HCC (5)
HCC GI (12)	0.150	0.085	0.0091	0.0001	0.0001	0.0001
HCC GII (11)
HCC GIII (10)
Tumor size
≤2 cm (10)	0.097	0.158	0.077	0.079	0.0001	0.0191
>2 cm (28)
Encapsulation
E-HCC (5)
Positive (27)	0.434	0.634	0.0491	0.0011	0.0111	0.0051
Negative (6)
Microvascular invasion
Positive (16)	0.413	0.339	0.0421	0.0001	0.0021	0.0001
Negative (22)
Major portal vein invasion
Positive (3)	0.271	0.489	0.739	0.0121	0.0501	0.0281
Negative (35)
Intrahepatic metastasis
Positive (7)	0.798	0.189	0.158	0.0001	0.0101	0.0501
Negative (31)
Tumor stage I (25)
II (10)	0.800	0.816	0.128	0.0001	0.0021	0.0001
III (3)

E: early; G: Edmondson-Steiner’s grade; ( ): number of cases; P: Jonckheere-Terpstra test, Mann–Whitney test, or Kruskal–Wallis test;

¹Value was statistically significant.