Is the expression of γ-glutamyl transpeptidase messenger RNA an indicator of biological behavior in recurrent hepatocellular carcinoma?

Table 1 Demography

Characteristics	Total (n = 40)
Age (years)	52.4 ± 16.6 [16-78]
Male26 (65%)
Liver cirrhosis	33 (82.5%)
Child-Pugh class A:B	32:8 (80%:20%)
Tumor size
Small ( < 3 cm)	6 (15.0%)
Median (3-10 cm)	23 (57.5%)
Large ( > 10 cm)	11 (27.5%)
HBsAg (+)	22 (55.0%)
Anti-HCV (+)	15 (37.5%)
AFP
Below 20 µg /L	17 (42.5%)
Over 1000 µg/L	14 (35.0%)
Capsule
Complete	18 (45.0%)
Incomplete or absent	22 (55.0%)
Edmondson-Steiner grade
I & II	19 (47.5%)
III & IV	21 (52.5%)
Vascular permeation	17 (42.5%)
Satellite formation	19 (47.5%)
Multiple HCC	13 (32.5%)
Major resection	32 (80.0%)
Minor resection	8 (20.0%)

Table 2 Nucleotide sequences of the primer sets and specific oligonucleotide probes to each type of γ-GTP 5’-noncoding mRNA

Type of γ-GTP mRNA or	Primers probes	Nucleotide sequences
Primer sets
A type	Sense	5’-CAC AGG GGA CAT ACA GTG AG-3’
	Antisense	5’-GAA ATA GCT GAA GCA CGC GC-3’
B type	Sense	5’-GGA TTC TCC CAG AGA TTG CC-3’
	Antisense	5’-GAA GGT CAA GGG AGG TTA CC-3’
C type	Sense	5’-GCC CAG AAG TGA GAG CAG TT-3’
	Antisense	5’-TCC AGA AAG CAG CTA GAG GG-3’
Oligonucleotide probes
A type		5’-GGG CAG GGC TTG GTG AAT GGT
		AGC TGT GAT TAT CAT CAT G-3’
B type		5’-CGC AGG ATG GTG TGC GAG GAC
		CCC GAG CTG GTG TTC CAG GC-3’
C type		5’-ATA GAG ACA CCG ATT CCT GGA
		GGT CCA AAG AGC CTC AGG A-3’

Table 3 Frequency of γ-GTP mRNA in liver

Tissues	mRNA types (%)
	A	A+B	B	A+B or B	C
HCC
Cancerous	1 (2.5)	15 (37.5)	24 (60.0)	39 (97.5)	0 (0)
Non-Cancerous	12 (30.0)	12 (30.0)	16 (40.0)	28 (70.0)	0 (0)
	A	B		C
Control	15 (100.0)	1 (6.7)			1 (6.7)

Table 4 Correlation between type B γ-GTP mRNA and clinical parameters

Parameters	Cases	Type B/A+B(%)
		HCC liver	Non HCC liver
Male	26	23 (88.5)	20 (76.9)
Female	14	10 (71.4)	8 (43.9)
HBsAg (+)	22	19 (86.4)	17 (60.7)
(-)	18	14 (77.8)	11 (39.3)
HCV (+)	15	14 (93.3)	14 (93.3)b
(-)	25	19 (76.0)	14 (56.0)b
Child A	32	25 (75.8)	21 (65.6)
Child B	8	8 (100.0)	7 (87.5)
AFP (ng/mL) < 20	17	11 (64.7)a	9 (32.1)c
= 20	23	22 (95.6)a	19 (82.6)c

Denotes:

^aP = 0.0295;

^bP = 0.0151;

^cP = 0.0430.

Table 5 Correlation between types of γ-GTP mRNA and patho-logical parameters

Parameters	Cases	Type B/A+B(%)
		HCC liver	Non HCC liver
Cirrhosis (+)	33	27 (81.8)	22 (66.7)
(-)	7	6 (85.7)	6 (85.7)
Size< 3 cm	6	6 (100.0)	4 (66.7)
3-10 cm	11	7 (63.6)	5 (45.4)
=10 cm	23	20 (86.9)	19 (82.6)
Differentiated grade I/II	19	14 (73.7)	10 (52.6)b
III/IV	21	19 (90.5)	18 (85.7)b
Complete capsule (+)	18	13 (72.2)	8 (44.4)c
(-)	22	20 (90.9)	20 (90.9)c
Vascular permeation (+)	17	16 (94.1)	16 (94.1)d
(-)	23	17 (73.9)	12 (52.2)d
Daughter nodules (+)	19	19 (100.0)a	18 (94.7)e
(-)	21	14 (66.7)a	10 (47.6)e
Multi-centric (+)	13	12 (92.3)	12 (92.3)
(-)	27	21 (77.8)	16 (59.2)

Denotes:

^aP = 0.0089;

^bP = 0.0226;

^cP = 0.0014;

^dP = 0.0042;

^eP = 0.0012.

Table 6 Correlation between γ-GTP mRNA and their outcome

γ-GTP mRNA	Cases	Recurrence (%)	Death (%)
Type B in HCC liver (+)	33	21(63.6)a	15(45.5)b
(-)	7	1(14.3)a	0(0)b
Type B in non-HCC liver (+)	28	22(78.6)c	15(53.6)d
(-)	12	0(0)c	0(0)d

Denotes:

^aP = 0.0328;

^bP = 0.0328;

^cP: < 0.0001;

^dP: 0.0011

Table 7 Significant factors in recurrence and death

Parameters	Recurrence		Death
	UV	MV	UV	MV
Sex	n.s.	n.s.	n.s.	n.s.
Age	n.s.	n.s.	n.s.	n.s.
Child-Pugh’s classification: A vs. B	n.s.	n.s.	n.s.	n.s.
HBsAg (+)	n.s.	n.s.	0.0166	n.s.
HCV (+)	n.s.	n.s.	n.s.	n.s.
AFP < 20 vs. 20-1000	0.0001	0.0108	0.0006	0.0141
γ-GTP mRNA (+)	n.s.	n.s.	n.s.	n.s.
Pathology of HCC:
Size: <3 cm vs. 3-10 cm	0.0213	n.s.	0.0080	n.s.
Cirrhosis	n.s.	n.s.	n.s.	n.s.
Edmondson-Steiner grade	0.0001	n.s.	0.0007	n.s.
Complete capsule (+)	< 0.0001	0.0084	< 0.0001	0.0130
Vascular permeation (+)	0.0003	n.s.	0.0001	0.0053
Daughter nodules (+)	0.0549	n.s.	0.0550	n.s.
Multicentric (+)	0.0094	n.s.	0.0119	n.s.
Type B γ-GTP mRNA in HCC liver	0.0003	0.0107	0.0046	n.s.
Type B γ-GTP mRNA in non-HCC liver	0.0001	n.s.	0.0007	n.s.

Denotes: n.s. = not significant.