Amplification of chromosome 21q22.3 harboring trefoil factor family genes in liver fluke related cholangiocarcinoma is associated with poor prognosis

Table 1 Locations and sequences of microsatellite markers, trefoil factor family genes and reference loci

Primer name	Chromosome	Product size (bp)	Forward primer	Reverse primer
D21S1253	21q21.3	174-190	GAAGAATCTCCCGAACCAGG	AAGACCAGTGTTATTCAGAGCC
D21S1255	21q22.2	112-126	AGCTCTTTATTTTGCCACATAG	CTGCATGTTTGCCTGG
D21S1893	21q22.2	111-119	GTATGCACACCACACGG	TAACAAAATCCGCCACG
D21S1890	21q22.3	143-173	GGTCTGACCACAGATTTCC	AAAAACACTCTGAACGATTAAGG
Trefoil factor family 1	21q22.3	219	CAGGGATCTGCCTGCATC	ATCGATCTCTTTTAATTTTTAGGCC
Trefoil factor family 2	21q22.3	123	GAAGAATCTCCCGAACCAGG	GTCACACTTCAAAAACTAGAGG
Trefoil factor family 3	21q22.3	129	CAGGCACTGTTCATCTCAGC	TATTCGTTAAGACATCAGGCTCC
β-actin	7p15	375	TCACCCACACTGTGCCCATCTACGA	CAGCGGAACCGCTCATTGCCAATGG
GAPDH1	12p13	250	ACAGTCCATGCCATCACTGCC	GCCTGCTTCACCACCTTCTTG

¹Glyceraldehyde-3-phosphate dehydrogenase.

Table 2 Clinicopathological parameters of patients with and without allelic imbalance

Clinical parameters	n	Gene/microsatellite makersD21S1893, D21S1890 and TFF
		Normaln (%)	Amplificationn (%)	Deletionln (%)	P value
Age (yr)	80
≤ 54	39	18 (46)	20 (51)	1 (3)	NS
＞ 54	41	15 (36)	22 (54)	4 (10)
Gender	80
Male	57	22 (39)	32 (56)	3 (5)	NS
Famale	23	11 (48)	10 (43)	2 (9)
Histological type	80
Non-Papillary adenocarcinoma	58	27 (46)	30 (52)	1 (2)	0.023
Papillary adenocarcinoma	22	6 (27)	12 (55)	4 (18)
Staging	80
II & III	10	5 (50)	5 (50)	-	NS
IVA & IVB	70	28 (40)	37 (52.8)	5 (7.2)
Blood vessel invasion	79
Non-invasion	28	8 (29)	16 (57)	4 (14)	NS
Invasion	51	24 (47)	26 (51)	1 (2)
Lymphatic invasion	79
Non-invasion	15	4 (27)	11 (73)	-	NS
Invasion	64	28 (43)	31 (49)	5 (8)
Nerve invasion	79
Non-invasion	36	14 (39)	19 (53)	3 (8)	NS
Invasion	43	18 (42)	23 (54)	2 (4)
Survival time (wk)	69				0.012
Mean		54.12	51.7	124.82
Median		49	27.4	84.28
Minimum-maximum		7.14-119.28	4.85-242.85	67.71-143.85

Table 3 Univariate and multivariate analysis of overall survival in cholangiocarcinoma

Variable	n	Univariate		Multivariate
		Relative risk(95%CI)	P value	Relative risk(95%CI)	P value
Age (yr)	69	0.831 (0.483-1.430)	NS
≤ 54	34
＞ 54	35
Gender	69	0.443 (0.231-0.849)	0.014	0.628 (0.312-1.263)	NS
Male	49
Famale	20
Histological type	69	0.622 (0.332-1.167)	NS
Non-Papillary adenocarcinoma	50
Papillary adenocarcinoma	19
Staging	69	2.265 (0.899-5.708)	0.083	3.320 (1.270-8.681)	0.014
II & III	9
IVA & IVB	60
Blood vessel invasion	69	2.108 (1.139-3.902)	0.018	2.183 (1.088-4.382)	0.028
Non-invasion	23
Invasion	46
Lymphatic invasion	69	0.981 (0.479-2.012)	NS
Non-invasion	12
Invasion	57
Nerve invasion	69	1.129 (0.654-1.950)	NS
Non-invasion	32
Invasion	37
D21S1893, D21S1890, and TFF	69		0.026		0.002
Normal	30	Reference		Reference
Amplification	35	1.707 (0.974-2.991)	NS	2.473 (1.342-4.557)	0.004
Deletion	4	0.187 (0.025-1.394)	NS	0.224 (0.029-1.701)	NS