Altered promoter usage characterizes monoallelic transcription arising with ERBB2 amplification in human breast cancers.
Analysis of a collection of human breast cancers (n = 150), enriched in ERBB2-positive cases (n = 57) and involving tumor genotyping relative to population-matched blood genotyping (n = 749) for a common ERBB2 single nucleotide polymorphism Ala(G)1170Pro(C), revealed that ERBB2 amplification in breast cancer is invariably monoallelic. Analysis of paired breast cancer and blood samples from informative (G1170C heterozygotic) ERBB2-positive (n = 12) and ERBB2-negative (n = 17) cases not only confirmed monoallelic amplification and ERBB2 transcriptional overexpression but also revealed that most low ERBB2 expressing breast cancers (12/17) exhibit unbalanced allelic transcription, showing 3-fold to nearly 5,000-fold preferential expression from one of two inherited alleles. To explore cis-acting transcriptional mechanisms potentially selected during ERBB2 amplification, levels of four different ERBB2 transcript variants (5.2, 4.7, 2.1, and 1.4 kb) were correlated with total (4.6 kb) ERBB2 mRNA levels in ERBB2-positive (n = 14) versus ERBB2-negative (n = 43) primary breast cancers. Relative expression of only the 2.1 kb extracellular domain-encoding splice variant and a 4.7 kb mRNA variant that uses an alternative start site were significantly increased in association with ERBB2-positivity, implicating altered promoter usage and selective transcript regulation within the ERBB2 amplicon. Altogether, these findings provide new mechanistic insights into the development of ERBB2-positive breast cancer and strong rationale for delineating candidate cis-acting regulatory elements that may link allele-specific ERBB2 transcription in premalignant breast epithelia with subsequent development of breast cancers bearing monoallelic ERBB2 amplicons.
Benz
Christopher C
CC
Buck Institute for Age Research, Novato, CA 94945, USA. cbenz@buckinstitute.org
Fedele
Vita
V
Xu
Fan
F
Ylstra
Bauke
B
Ginzinger
David
D
Yu
Mamie
M
Moore
Dan
D
Hall
Rayna Kneuper
RK
Wolff
Daynna J
DJ
Disis
Mary L
ML
Eppenberger-Castori
Serenella
S
Eppenberger
Urs
U
Schittulli
Francesco
F
Tommasi
Stefania
S
Paradiso
Angelo
A
Scott
Gary K
GK
Albertson
Donna G
DG
D000483:Alleles; D017398:Alternative Splicing; D001943:Breast Neoplasms / Q000235:genetics*; D005260:Female; D005784:Gene Amplification; D018628:Gene Dosage; D015972:Gene Expression Regulation, Neoplastic; D018734:Genes, erbB-2 / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008297:Male; D009361:Neoplasm Invasiveness / Q000235:genetics; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014158:Transcription, Genetic