S100A6 binds to annexin 2 in pancreatic cancer cells and promotes pancreatic cancer cell motility.
BACKGROUND
High levels of S100A6 have been associated with poor outcome in pancreatic cancer patients. The functional role of S100A6 is, however, poorly understood.


METHODS
Immunoprecipitation followed by two-dimensional gel electrophoresis and mass spectrometry were undertaken to identify S100A6 interacting proteins in pancreatic cancer cells. Immunohistochemistry and coimmunofluorescence were performed to examine expression or colocalisation of proteins. siRNA was used to deplete specific proteins and effects on motility were measured using Boyden Chamber and wound healing assays.


RESULTS
Our proteomic screen to identify S100A6 interacting proteins revealed annexin 11, annexin 2, tropomyosin beta and a candidate novel interactor lamin B1. Of these, annexin 2 was considered particularly interesting, as, like S100A6, it is expressed early in the development of pancreatic cancer and overexpression occurs with high frequency in invasive cancer. Reciprocal immunoprecipitation confirmed the interaction between annexin 2 and S100A6 and the proteins colocalised, particularly in the plasma membrane of cultured pancreatic cancer cells and primary pancreatic tumour tissue. Analysis of primary pancreatic cancer specimens (n=55) revealed a strong association between high levels of cytoplasmic S100A6 and the presence of annexin 2 in the plasma membrane of cancer cells (P=0.009). Depletion of S100A6 was accompanied by diminished levels of membrane annexin 2 and caused a pronounced reduction in the motility of pancreatic cancer cells.


CONCLUSIONS
These findings point towards a functional role for S100A6 that may help explain the link between S100A6 expression in pancreatic cancer and aggressive disease.
Nedjadi
T
T
Division of Surgery and Oncology, University of Liverpool, Liverpool, UK.
Kitteringham
N
N
Campbell
F
F
Jenkins
R E
RE
Park
B K
BK
Navarro
P
P
Ashcroft
F
F
Tepikin
A
A
Neoptolemos
J P
JP
Costello
E
E
D017306:Annexin A2 / Q000032:analysis / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000502:physiology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D003593:Cytoplasm / Q000737:chemistry; D006801:Humans; D047468:Immunoprecipitation; D010190:Pancreatic Neoplasms / Q000737:chemistry / Q000473:pathology*; D034622:RNA Interference; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins / Q000502:physiology*
C472822:ANXA2 protein, human; D017306:Annexin A2; D018797:Cell Cycle Proteins; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; C055351:S100A6 protein, human