Differential roles of EPS8 in carcinogenesis: Loss of protein expression in a subset of colorectal carcinoma and adenoma

Table 1 Epidermal growth factor receptor pathway substrate 8 genomic primers (ENST00000389337)

Primer	Sequence	Primer for sequence	Tm	PCR fragment (bp)
EPS8ex1F	tcctggcagcaacacatatt	F	59	227
EPS8ex1R	ccaaatcaaattcccccaaa		62
EPS8ex2F	aacccaacacaatgaccttttt		60	251
EPS8ex2R	tcactgcctcattccaaaca	R	60
EPS8ex3F	gagatagccacatgataccaaca		59	195
EPS8ex3R	tgttcctcaagggtcactctaaa	F	60
EPS8ex4F	tctttttcctttttgccaat		56	280
EPS8ex4R	ttccatccattttcaaacaatc	R	58
EPS8ex5F	gattgtttgaaaatggatggaa	F	59	261
EPS8ex5R	aaagctcccagacaatctgc		59
EPS8ex6F	tcagacaaggaacaatcccttt	F	60	251
EPS8ex6R	tttttctaactctttggggaaaaa		60
EPS8ex7F	agtaccacaagttgagttaattgat	F	55	264
EPS8ex7R	tcccaacccaaagtaagtgttc		60
EPS8ex8F	ggcaaatggtcctccttttt	F	60	206
EPS8ex8R	ccagtgatctaaaggcgactc		59
EPS8ex9F	tgggctgcttccttttctaa	F	60	280
EPS8ex9R	ctggagatctaaccaggcatt		58
EPS8ex10F	cctctcctctcgcttatttca	F	58	234
EPS8ex10R	cacacccccacaaaatctat		57
EPS8ex11F	gaccgtcccctctgtgtcta	F	60	229
EPS8ex11R	ccagacagacacttggggtta		60
EPS8ex12F	cttgtttttgccatgggttt	F	60	265
EPS8ex12R	aaggcattataggtggtaaatgct		59
EPS8ex13F	tatgccttcattccctcctg	F	60	297
EPS8ex13R	tgaataaaatgagaacttgcaatca		60
EPS8ex14F	tgacctgagtgctgattcaaa	F	59	274
EPS8ex14R	gacactgtcacctctgttagcac		59
EPS8ex15F	ctttaggaagagctagcagaat		54	250
EPS8ex15R	aatacttttgaaggaaagtttagttat	R	54
EPS8ex16F	gggaacttctttcgtagaatgg	F	59	267
EPS8ex16R	aagagtgataacttcgtaaatgtgt		56
EPS8ex17F	aaagtataatttgttttcctagcc	F	55	315
EPS8ex17R	tgcctccctgggaaacttac		61
EPS8ex18F	ggggttctagagagggtgatgt	F	61	292
EPS8ex18R	tgttgtacacacagaattgcaaag		60
EPS8ex19F	tttctcctttgtttgtaggcaat		59	256
EPS8ex19R	aatagttgtttccagagctttcaa	R	59
EPS8ex20F	gcagcctgcacaagtcagta	F	60	203
EPS8ex20R	aatgccaaaaacaatggagtt		59

EPS8: Epidermal growth factor receptor pathway substrate 8; PCR: Polymerase chain reaction; F: Forward; R: Reverse.

Table 2 Analysis of epidermal growth factor receptor pathway substrate 8 gene status and protein expression in uncultured tumor specimens n (%)

	Total number	Membranousβ-catenin	LOH1	EPS8 protein loss	EPS8 mutation2	Methylation3
Sporadic carcinomas	22	10/22 (45)	7/19 (37)	11/22 (50)	0/11	0/7
Sporadic adenomas	15	4/10 (40)	6/15 (40)	4/14 (29)	0/4	ND
FCC-X	21	11/18 (61)	7/17 (41)	4/15 (27)	ND	ND
Total	58		20/51 (39)	19/51 (37)	0/15	0/7

¹Based on informative (i.e., constitutionally heterozygous) cases;

²Cases with protein loss by immunohistochemistry and/or presence of loss of heterozygosity (LOH) were selected for this analysis;

³Cases were selected on the basis of immunohistochemical protein loss without LOH. ND: Not done; FCC-X: Familial colon cancer type X; EPS8: Epidermal growth factor receptor pathway substrate 8.