Ezrin expression and cell survival regulation in colorectal cancer.
Colorectal cancer (CRC) is the second largest cause of cancer deaths in the United States. A key barrier that prevents better outcomes for this type of cancer as well as other solid tumors is the lack of effective therapies against the metastatic disease. Thus there is an urgent need to fill this gap in cancer therapy. We utilized a 2D-DIGE proteomics approach to identify and characterize proteins that are differentially regulated between primary colon tumor and liver metastatic deposits of the IGF1R-dependent GEO human CRC xenograft, orthotopically implanted in athymic nude mice that may serve as potential therapeutic targets against CRC metastasis. We observed increased expression of ezrin in liver metastasis in comparison to the primary colonic tumor. Increased ezrin expression was further confirmed by western blot and microarray analyses. Ezrin, a cytoskeletal protein belonging to Ezrin-Radixin-Moesin (ERM) family plays important roles in cell motility, invasion and metastasis. However, its exact function in colorectal cancer is not well characterized. Establishment of advanced GEO cell lines with enhanced liver-metastasizing ability showed a significant increase in ezrin expression in liver metastasis. Increased phosphorylation of ezrin at the T567 site (termed here as p-ezrin T567) was observed in liver metastasis. IHC studies of human CRC patient specimens showed an increased expression of p-ezrin T567 in liver metastasis compared to the primary tumors of the same patient. Ezrin modulation by siRNA, inhibitors and T567A/D point mutations significantly downregulated inhibitors of apoptosis (IAP) proteins XIAP and survivin that have been linked to increased aberrant cell survival and metastasis and increased cell death. Inhibition of the IGF1R signaling pathway by humanized recombinant IGF1R monoclonal antibody MK-0646 in athymic mouse subcutaneous xenografts resulted in inhibition of p-ezrin T567 indicating ezrin signaling is downstream of the IGF1R signaling pathway. We identified increased expression of p-ezrin T567 in CRC liver metastasis in both orthotopically implanted GEO tumors as well as human patient specimens. We report for the first time that p-ezrin T567 is downstream of the IGF1R signaling and demonstrate that ezrin regulates cell survival through survivin/XIAP modulation.
Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, 985950 Nebraska Medical Center, Omaha, NE 68198-5950, United States.
Premila D
PD
Leiphrakpam
Department of Surgery, University of New Mexico Health Science Center, 1 University of New Mexico, Albuquerque, NM 87131-0001, United States.
Ashwani
A
Rajput
Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, 985950 Nebraska Medical Center, Omaha, NE 68198-5950, United States.
Michelle
M
Mathiesen
Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, 985950 Nebraska Medical Center, Omaha, NE 68198-5950, United States.
Ekta
E
Agarwal
Department of Pathology and Microbiology, University of Nebraska Medical Center, 983515 Nebraska Medical Center, Omaha, NE 68198-3135, United States.
Audrey J
AJ
Lazenby
Department of Surgical Oncology, University of Nebraska Medical Center, 984533 Nebraska Medical Center, Omaha, NE 68198-4533, United States.
Chandrakanth
C
Are
Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, 985950 Nebraska Medical Center, Omaha, NE 68198-5950, United States. Electronic address: mbrattain@unmc.edu.
Michael G
MG
Brattain
Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, 985950 Nebraska Medical Center, Omaha, NE 68198-5950, United States. Electronic address: schowdhury@unmc.edu.
Sanjib
S
Chowdhury
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D061067:Antibodies, Monoclonal, Humanized; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival; D015179:Colorectal Neoplasms / Q000378:metabolism* / Q000473:pathology; D003598:Cytoskeletal Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism; D008113:Liver Neoplasms / Q000378:metabolism / Q000556:secondary; D051379:Mice; D008819:Mice, Nude; D010766:Phosphorylation; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D017526:Receptor, IGF Type 1 / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D000077022:Survivin; D014183:Transplantation, Heterologous; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000378:metabolism; D000099149:Ezrin
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D003598:Cytoskeletal Proteins; D051034:Inhibitor of Apoptosis Proteins; D034741:RNA, Small Interfering; D017526:Receptor, IGF Type 1; D000077022:Survivin; D051636:X-Linked Inhibitor of Apoptosis Protein; D000099149:Ezrin; C107489:BIRC5 protein, human; C569480:dalotuzumab
2D-DIGE proteomics; Cell survival signaling; Colorectal cancer metastasis; Ezrin; IGF1R signaling; XIAP and survivin