Increased expression of Cyr61 (CCN1) identified in peritoneal metastases from human pancreatic cancer.
BACKGROUND
Identification of extracellular matrix proteins (ECM) associated with tumor cell metastasis may generate targets for future therapy against pancreatic cancer metastases. We hypothesized that comparison of ECM-associated gene expression in primary and metastatic pancreatic tumors would identify ECM proteins associated with pancreatic metastasis.


METHODS
A clinically relevant model of pancreatic cancer was used to generate RNA from primary and metastatic tumors; it was evaluated by microarray analysis with subsequent cluster analysis. Target genes (Cyr61 and integrins alpha(v) and beta(3)) identified by microarray analysis were confirmed by reverse transcription polymerase chain reaction and immunohistochemistry analysis.


RESULTS
Peritoneal metastases at sites distant from the primary tumor were present in all animals bearing orthotopic tumors. High-density microarray comparison of gene expression in metastases versus primary pancreatic tumors identified a greater than twofold increase in the expression of Cyr61, a secreted matricellular protein that binds to integrins. Reverse transcription polymerase chain reaction confirmed the microarray results, and immunohistochemistry analysis demonstrated increased Cyr61 protein and persistent alpha(v)beta(3) expression in peritioneal metastases. Additionally, immunohistochemistry demonstrated increased collocalization of Cyr61 and alpha(v) in metastases relative to primary tumor.


CONCLUSIONS
The ECM protein Cyr61 shows increased expression in metastatic lesions in a clinically relevant model of pancreatic adenocarcinoma. Protein analysis confirms the microarray results and collocalization of Cyr61, and alpha(v) suggests that interaction between Cyr61 and alpha(v)beta(3) promotes formation of peritoneal metastases.
Holloway
Shane E
SE
Division of Surgical Oncology, Department of Surgery, Hamon Center for Therapeutic Oncology, University of Texas Southwestern Medical Center, Dallas, TX 75390-8593, USA.
Beck
Adam W
AW
Girard
Luc
L
Jaber
M Raffat
MR
Barnett
Carlton C
CC
Brekken
Rolf A
RA
Fleming
Jason B
JB
D000230:Adenocarcinoma / Q000235:genetics / Q000378:metabolism / Q000556:secondary*; D000818:Animals; D000912:Antibodies, Neoplasm / Q000276:immunology; D016000:Cluster Analysis; D055515:Cysteine-Rich Protein 61; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D010534:Peritoneal Neoplasms / Q000235:genetics / Q000378:metabolism / Q000556:secondary*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D000912:Antibodies, Neoplasm; C501848:CCN1 protein, human; C491850:CCN1 protein, mouse; D055515:Cysteine-Rich Protein 61; D016326:Extracellular Matrix Proteins; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm