Human antimicrobial protein hCAP18/LL-37 promotes a metastatic phenotype in breast cancer.
BACKGROUND
Human cathelicidin antimicrobial protein, hCAP18, and its C-terminal peptide LL-37 is a multifunctional protein. In addition to being important in antimicrobial defense, it induces chemotaxis, stimulates angiogenesis and promotes tissue repair. We previously showed that human breast cancer cells express high amounts of hCAP18, and hypothesised that hCAP18/LL-37 may be involved in tumour progression.


METHODS
hCAP18 mRNA was quantified in 109 primary breast cancers and compared with clinical findings and ERBB2 mRNA expression. Effects of exogenous LL-37 and transgenic overexpression of hCAP18 on ErbB2 signalling were investigated by immunoblotting using extracts from breast cancer cell lines ZR75-1 and derivatives of MCF7. We further analysed the impact of hCAP18/LL-37 on the morphology of breast cancer cells grown in soft agar, on cell migration and on tumour development in severe combined immunodeficiency (SCID) mice.


RESULTS
The expression of hCAP18 correlated closely with that of ERBB2 and with the presence of lymph node metastases in oestrogen receptor-positive tumours. hCAP18/LL-37 amplified Heregulin-induced mitogen-activated protein kinase (MAPK) signalling through ErbB2, identifying a functional association between hCAP18/LL-37 and ErbB2 in breast cancer. Treatment with LL-37 peptide significantly stimulated the migration of breast cancer cells and their colonies acquired a dispersed morphology indicative of increased metastatic potential. A truncated version of LL-37 competitively inhibited LL-37 induced MAPK phosphorylation and significantly reduced the number of altered cancer cell colonies induced by LL-37 as well as suppressed their migration. Transgenic overexpression of hCAP18 in a low malignant breast cancer cell line promoted the development of metastases in SCID mice, and analysis of hCAP18 transgenic tumours showed enhanced activation of MAPK signalling.


CONCLUSIONS
Our results provide evidence that hCAP18/LL-37 contributes to breast cancer metastasis.
Weber
Günther
G
GICC, CNRS UMR 6239, Université François Rabelais, Avenue Monge, Tours, France. gunther.weber@univ-tours.fr
Chamorro
Clara Ibel
CI
Granath
Fredrik
F
Liljegren
Annelie
A
Zreika
Sami
S
Saidak
Zuzana
Z
Sandstedt
Bengt
B
Rotstein
Samuel
S
Mentaverri
Romuald
R
Sánchez
Fabio
F
Pivarcsi
Andor
A
Ståhle
Mona
M
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000502:physiology*; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D016022:Case-Control Studies; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D003114:Colony-Forming Units Assay; D005260:Female; D006801:Humans; D008207:Lymphatic Metastasis; D051379:Mice; D016513:Mice, SCID; D008875:Middle Aged; D020928:Mitogen-Activated Protein Kinases; D010641:Phenotype; D010766:Phosphorylation; D012333:RNA, Messenger / Q000378:metabolism; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics*; D011960:Receptors, Estrogen / Q000378:metabolism; D015398:Signal Transduction; D054804:Cathelicidins
D023181:Antimicrobial Cationic Peptides; D020928:Mitogen-Activated Protein Kinases; D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases; D011960:Receptors, Estrogen; D054804:Cathelicidins