Identification of biomarkers of human pancreatic adenocarcinomas by expression profiling and validation with gene expression analysis in endoscopic ultrasound-guided fine needle aspiration samples

Table 1 Sense (S) and antisense (AS) primers used in quantitative RT-PCR analyses

Primer	Exon	Sequence (5’-3’)	Amplicon
LCN2 (Lipocalin2) S	1	TGATCCCAGCCCCACCT	74 bp
LCN2 (Lipocalin2) AS	2	CCACTTCCCCTGAATTGGT
PLAT (tPA) S	2	TGGAGAGAAAACCTCTGCGAG	72 bp
PLAT (tPA) AS	3	CCATGATTGCTTCACAGCGT
KRT7 (Keratin7) S	7	CTCTGTGATGAATTCCACTGGTG	72 bp
KRT7 (Keratin7) AS	8	CCCATGGTTCCCCCGA
18S S		AAACGGCTACCACATCCAAG	155 bp
18S AS		CCTCCAATGGATCCTCGTTA

Table 2 Significantly overexpressed genes (over 2-fold) in malignant pancreas

Gene symbol	Gene description	Mean fold expression ratios
		Malignant	PCL/	PT/
		Panc/others	CCL	NormPanc
PLAT	Plasminogen activator, tissue-type	6.22	4.36	6.74
KRT7	Keratin 7	5.72	4.92	3.97
CD74	CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated)	5.44	1.61	8.00
MMP7	Matrix metalloproteinase 7 (matrilysin, uterine)	5.36	3.73	5.70
LCN2	Lipocalin 2 (oncogene 24p3), NGAL	5.25	8.32	3.36
HLA-G	HLA-G histocompatibility antigen, class I, G	4.04	1.57	4.32
IGHG3	Immunoglobulin heavy constant gamma 3	3.95	0.94	11.81
HLA-DRA	Major histocompatibility complex, class II, DR alpha	3.70	1.57	7.01
TIMP1	Tissue inhibitor of metalloproteinase 1 (erythroid potentiating activity, collagenase inhibitor)	3.50	1.61	9.20
ITGA3	Integrin, alpha 3 (antigen CD49C, alpha 3 subunit of VLA-3 receptor)	3.48	3.00	2.40
ITGB4	Integrin, beta 4	3.36	1.16	4.34
KRT19	Keratin 19	3.27	1.34	2.98
CTSD	Cathepsin D (lysosomal aspartyl protease)	3.21	2.75	2.89
CASP4	Caspase 4, apoptosis-related cysteine protease	3.10	2.43	1.80
IGFBP3	Insulin-like growth factor binding protein 3	3.08	3.53	1.87
PLAU	Plasminogen activator, urokinase	3.08	2.83	2.01
MMP11	Matrix metalloproteinase 11 (stromelysin 3)	2.96	1.11	7.22
DTR	Diphtheria toxin receptor (heparin-binding epidermal growth factor-like growth factor)	2.84	1.74	3.33
CDKN1A	Cyclin-dependent kinase inhibitor 1A (p21, Cip1)	2.76	1.65	3.17
LAMA4	Laminin, alpha 4	2.74	2.14	3.30
ITGB8	Integrin, beta 8	2.59	2.31	4.44
IFITM1	Interferon induced transmembrane protein 1 (9-27)	2.41	5.47	7.10
AXL	AXL receptor tyrosine kinase	2.35	1.63	2.91
ITGAE	Integrin, alpha E (antigen CD103, human mucosal lymphocyte antigen 1; alpha polypeptide)	2.31	1.58	3.56
CD59	CD59 antigen p18-20 (antigen identified by monoclonal antibodies 16.3A5, EJ16, EJ30, EL32 and G344)	2.26	2.11	2.95
KRT10	Keratin 10 (epidermolytic hyperkeratosis; keratosis palmaris et plantaris)	2.09	2.18	1.60
CYR61	Cysteine-rich, angiogenic inducer, 61	2.03	2.41	1.22
PTGES	Prostaglandin E synthase	2.03	2.84	1.11
HIF1A	Hypoxia-inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)	2.01	1.24	2.87

Table 3 Relative mRNA levels presented as 2^{[Ct(18S)-Ct(gene of interest)]} (mean x 10^-2±SEM)

Gene	n	KRT7	LCN2	PLAT
Pancreatic tumors	6	1.14. ± 0.47	3.58 ± 0.65a	2.34 ± 0.89a
Pancreatic cell lines	6	79.62 ± 90.97	188.99 ± 81.13a	13.51 ± 4.67a
Normal pancreas	3	0.42 ± 0.23	1.05 ± 0.85	0.14 ± 0.08

^aP < 0.05, vs normal pancreas.