Prognostic Impact of mRNA Expression Levels of HER1–4 (ERBB1–4) in Patients with Locally Advanced Rectal Cancer

Table 1

Primer/probe sets used for amplification of the target and reference genes.

Gene	Oligo ID	Oligo sequences 5′-3′	Label probe	Amplicon length (nts)	Assay location	PCR efficacy in %
MKI67	Forward	CGAGACGCCTGGTTACTATCAA	CY5-BHQ2	108	c. ex 2-3 NM_002417	98
	Reverse	GGATACGGATGTCACATTCAATACC
	Probe	ACGGTCCCCACTTTCCCCTGAGC
CALM2	Forward	GAGCGAGCTGAGTGGTTGTG	YY-BHQ1	72	c. ex 1-2 NM_001743	99.3
	Reverse	AGTCAGTTGGTCAGCCATGCT
	Probe	TCGCGTCTCGGAAACCGGTAGC
EGFR (ERBB1)	Forward	CGCAAGTGTAAGAAGTGCGAA	FAM-BHQ1	93	NM_201283	89.1
	Reverse	CGTAGCATTTATGGAGAGTGAGTCT
	Probe	CCTTGCCGCAAAGTGTGTAACGGAAT
ERBB2 (HER2)	Forward	TCTGGACGTGCCAGTGTGAA	CY5-BHQ2	61	M11730_gen	97.2
	Reverse	CCTGCTCCCTGAGGACACAT
	Probe	AGGCCAAGTCCGCAGAAGCCCT
ERBB3 (HER3)	Forward	CGGTTATGTCATGCCAGATACAC	ROX-BHQ2	81	NM_001982	96
	Reverse	GAACTGAGACCCACTGAAGAAAGG
	Probe	CTCAAAGGTACTCCCTCCTCCCGGG
ERBB4 (HER4)	Forward	GAGGCTGCTCAGGACCTAAGG	ATTO-BHQ1	75	NM_005235	92
	Reverse	GAGTAACACATGCTCCACTGTCATT
	Probe	CACAGACTGCTTTGCCTGCATGAATTTC

CY5 = cyanine dye 5, BHQ = black hole quencher; FAM = 6-carboxyfluorescein; ROX = carboxy-X-rhodamine; ATTO = fluorescent dye (ATTO-TEC GmbH, Siegen, Germany).