Annexin A2 mediates up-regulation of NF-κB, β-catenin, and stem cell in response to progastrin in mice and HEK-293 cells.
OBJECTIVE
Prograstrin induces proliferation in colon crypts by activating p65nuclear factor-κB (NF-κB) (p65) and β-catenin. We investigated whether Annexin A2 (AnxA2), a progastrin receptor, activates NF-κB and β-catenin in vivo.


METHODS
ANXA2-null (ANXA2(-/-)) and wild-type (ANXA2(+/+)) mice were studied, along with clones of progastrin-responsive HEK-293 cells that stably expressed full-length progastrin (HEK-mGAS) or an empty vector (HEK-C). Small interfering RNA was used to down-regulate AnxA2, p65NF-κB, and β-catenin in cells.


RESULTS
Proliferation and activation of p65 and β-catenin increased significantly in HEK-mGAS compared with HEK-C clones. HEK-mGAS cells had a 2- to 4-fold increase in relative levels of c-Myc, cyclooxygenase (COX)-2, CyclinD1, double cortin CAM kinase-like 1 (DCAMKL+1), and CD44, compared with HEK-C clones. Down-regulation of AnxA2 in HEK-mGAS clones reduced activation of NF-κB and β-catenin, as well as levels of DCAMKL+1. Surprisingly, down-regulation of β-catenin had no effect on activation of p65NF-κB, whereas down-regulation of p65 significantly reduced activation of β-catenin in HEK-mGAS clones. Loss of either p65 or β-catenin significantly reduced proliferation of HEK-mGAS clones, indicating that both factors are required for the proliferative effects of progastrin. Lengths of colon crypts and levels of p65, β-catenin, DCAMKL+1, and CD44 were significantly higher in ANXA2(+/+) mice compared with either ANXA2(-/-) mice given progastrin or ANXA2(+/+) and ANXA2(-/-) mice given saline.


CONCLUSIONS
AnxA2 expression is required for the biologic effects of progastrin in vivo and in vitro and mediates the stimulatory effect of progastrin on p65NF-κ, β-catenin, and the putative stem cell markers DCAMKL+1 and CD44. AnxA2 might therefore mediate the hyperproliferative and cocarcinogenic effects of progastrin.
Sarkar
Shubhashish
S
Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, Texas 77555-1043, USA.
Swiercz
Rafal
R
Kantara
Carla
C
Hajjar
Katherine A
KA
Singh
Pomila
P
D000818:Animals; D017306:Annexin A2 / Q000235:genetics / Q000378:metabolism*; D049109:Cell Proliferation; D003106:Colon / Q000737:chemistry / Q000378:metabolism; D019938:Cyclin D1 / Q000032:analysis; D051546:Cyclooxygenase 2 / Q000032:analysis; D000089742:Doublecortin-Like Kinases; D015536:Down-Regulation; D005755:Gastrins / Q000378:metabolism* / Q000494:pharmacology; D057809:HEK293 Cells; D006801:Humans; D018960:Hyaluronan Receptors / Q000032:analysis; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis; D051379:Mice; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism* / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000032:analysis; D016271:Proto-Oncogene Proteins c-myc / Q000032:analysis; D034741:RNA, Small Interfering / Q000378:metabolism; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism*; D015854:Up-Regulation; D051176:beta Catenin / Q000378:metabolism*
D017306:Annexin A2; D005755:Gastrins; D018960:Hyaluronan Receptors; D047908:Intracellular Signaling Peptides and Proteins; C489427:MYC protein, human; C489428:Myc protein, mouse; D016328:NF-kappa B; D011498:Protein Precursors; D016271:Proto-Oncogene Proteins c-myc; D034741:RNA, Small Interfering; D051176:beta Catenin; D019938:Cyclin D1; C013706:big gastrin; D051546:Cyclooxygenase 2; C117347:DCLK1 protein, human; D000089742:Doublecortin-Like Kinases; C485525:Dclk1 protein, mouse; D017346:Protein Serine-Threonine Kinases
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