Distinct expression patterns in hepatitis B virus- and hepatitis C virus-infected hepatocellular carcinoma

Table 1 Clincal data of patients with hepatocellular carcinoma

Case	Sex	Age	Hepatitis virus	Differentiated grade	TNM score
1	M	54	HBV	WD	T1N0M0
2	M	60	HCV	WD	T1N0M0
3	F	61	HBV	WD	T2N0M0
4	M	62	HBV	WD	T2N0M0
5	M	58	HBV	WD	T1N0M0
6	F	56	HCV	MD	T3N0M0
7	F	44	HBV	WD	T2N0M0
8	M	49	HCV	WD	T1N0M0
9	M	58	HBV	WD	T2N0M0
10	M	67	HCV	PD	T3N0M0
11	M	69	HBV	WD	T2N0M0
12	F	63	HCV	WD	T1N0M0
13	M	48	HCV	MD	T2N0M0
14	F	63	HBV	WD	T1N0M0
15	M	49	HCV	MD	T1N0M0
16	F	51	HBV	PD	T3N1M0
17	M	65	HCV	MD	T3N1M0
18	F	58	HBV	PD	T4N1M1
19	M	60	HBV	MD	T2N1M0
20	F	56	HCV	PD	T3N1M1
21	M	42	HCV	PD	T3N1M0
22	M	55	HBV	PD	T4N1M1
23	M	66	HBV	MD	T3N1M0
24	F	70	HCV	WD	T2N1M0
25	M	58	HBV	PD	T4N1M1
26	M	53	HCV	PD	T3N1M0
27	M	61	HBV	PD	T4N1M0
28	F	65	HBV	MD	T3N1M0
29	M	59	HCV	MD	T3N1M1
30	M	50	HBV	PD	T3N1M0
31	F	63	HCV	PD	T4N1M1
32	M	66	HCV	PD	T3N1M0

M: Male; F: Female; HBV: Hepatitis B virus infection; HCV: Hepatitis C virus infection; WD: Well differentiated HCC; MD: Moderately differentiated HCC; PD: Poorly differentiated HCC.

Table 2 Primers used to amplify cDNA at various genes in real time RT-PCR

Target gene	Objective	Forward primer sequence (5'-3')	Reverse primer sequence (3'-5')	Genebank accession no./Amplicon size
RPS5	qRT-PCR	GTATGCCGCCAAACGCTTC	CGCCTGTGAGCAGGTGTAT	NM_001009, 152 bp
KRT8	qRT-PCR	GGAGGCATCACCGCAGTTAC	GGTTGGCAATATCCTCGTACTGT	NM_002273, 637 bp
CFLAR	qRT-PCR	GACAGAGCTTCTTCGAGACAC	GCTCGGGCATACAGGCAAAT	AF009616, 116 bp
ATP5F1	qRT-PCR	ACTGGGCTTATCTTGTACGCT	GCAAAGTCTGCAACAAAGGGA	NM_001688, 131 bp
IGFBP2	qRT-PCR	GACAATGGCGATGACCACTCA	GCTCCTTCATACCCGACTTGA	NM_000597, 121 bp
MAP3K5	qRT-PCR	AAAAAGGCATTTGAATCTGAGCC	GCTTGAATGACTCTCATGTGGTC	NM_005923, 233 bp
MMP9	qRT-PCR	GGGACGCAGACATCGTCATC	TCGTCATCGTCGAAATGGGC	NM_004994, 139 bp
VIM	qRT-PCR	AGTCCACTGAGTACCGGAGAC	CATTTCACGCATCTGGCGTTC	AK093924, 98 bp
ACTB	qRT-PCR	CATGTACGTTGCTATCCAGGC	CTCCTTAATGTCACGCACGAT	NM_001101, 250 bp
GAPD	qRT-PCR	CAACTGGTCGTGGACAACCAT	GCACGGACACTCACAATGTTC	AC002389, 260 bp
CD58	qRT-PCR	CTCATGGGATTGTCCTATGGAGC	GCTTGGGATACAGGTTGTCAAA	NM_001779, 154 bp

Table 3 The 11 genes for which expression levels differed between HBV- and HCV-associated HCC

Gene name	Symbol1	Accession2	Fold change3	HBV:HCV4
7 genes up-regulated in HBV-associated HCC
Ribosomal protein S5	RPS5	NM_001009	6.35	2.38
Keratin 8	KRT8	NM_002273	5.68	3.19
CASP8 and FADD-like apoptosis regulator	CFLAR	Y14039	2.86	2.09
ATP synthase, H+transporting, mitochondrial F0 complex, subunit b, isoform 1	ATP5F1	X60221	4.11	3.52
Insulin-like growth factor binding protein 2 (36 kDa)	IGFBP2	NM_000597	3.37	2.49
Mitogen-activated protein kinase kinase kinase 5	MAP3K5	NM_005923	3.76	2.33
Matrix metalloproteinase 9 (gelatinase B, 92 kDa gelatinase, 92 kDa type IV collagenase)	MMP9	NM_004994	7.43	3.74
4 genes up-regulated in HCV-associated HCC
Vimentin	VIM	NM_003380	8.61	0.28
Actin-β	ACTB	X00351	4.13	0.37
Glyceraldehyde-3-phosphate_dehydrogenase	GAPD	NM_002046	5.27	0.29
CD58 antigen, (lymphocyte function-associated antigen 3)	CD58	NM_001779	4.68	0.31

¹Symbol in LocusLink database;

²GeneBank accession number;

³Fold change, ratio of mean expression values in HBV- or HCV-associated cases (cancer cells vs non-cancerous cells);

⁴HBV:HCV ratio of mean expression values (HBV positive cases to HCV positive cases).