Frequent loss of endothelin-3 (EDN3) expression due to epigenetic inactivation in human breast cancer

Breast Cancer Research

Table 3 Oligonucleotide primers used in this study

	Sequence (5'↔3')	TA, °C	Cycles	Product, base pairs
RT-PCR
GAPDH
	Forward: TGGTCACCAGGGCTGCTT	60	35	510
	Reverse: GTCTTCTGGGTGGCAGTGAT
Real-time PCR
GAPDH
	Forward: GAAGGTGAAGGTCGGAGTCA	58	40	108
	Reverse: TGGACTCCACGACGTACTCA
EDN1
	Forward: GCTCGTCCCTGATGGATAAA	58	40	216
	Reverse: TTCCTGCTTGGCAAAAATTC
EDN2
	Forward: TTGGACATCATCTGGGTGAA	58	40	229
	Reverse: CTGTAGTGGCCCCTGTCTTG
EDN3
	Forward: ATTGCCACCTGGACATCATT	58	40	179
	Reverse: GCAGGCCTTGTCATATCTCC
Methylation-specific PCR
EDN3-U
	Forward: TTTGGGAGGTGATTTTTAGTGTGTTT	60	35	144
	Reverse: ACCCATCCCTACACAAAACTAACCA
EDN3-M
	Forward: TGGGAGGCGATTTTTAGTGCGTTC	60	35	140
	Reverse: CCATCCCTACGCGAAACTAACCG

EDN, endothelin; EDN3-M, endothelin-3 methylated; EDN3-U, endothelin-3 unmethylated; GAPDH, glyceraldehyde-3-phosphate-dehydrogenase; PCR, polymerase chain reaction; RT-PCR, reverse transcription-polymerase chain reaction; TA, annealing temperature.

ISSN: 1465-542X