Abstract
Calcium deficiency causes abnormal colonic growth and increases colon cancer risk with poorly understood mechanisms. Here we elucidate a novel signaling mechanism underlying the Ca2+ deficiency-induced epithelial proliferation using a unique animal model. The zebrafish larval yolk sac skin contains a group of Ca2+-transporting epithelial cells known as ionocytes. Their number and density increases dramatically when acclimated to low [Ca2+] environments. BrdU pulse-labeling experiments suggest that low [Ca2+] stimulates pre-existing ionocytes to re-enter the cell cycle. Low [Ca2+] treatment results in a robust and sustained activation of IGF1R-PI3K-Akt signaling in these cells exclusively. These ionocytes specifically express Igfbp5a, a high-affinity and specific binding protein for insulin-like growth factors (IGFs) and the Ca2+-selective channel Trpv5/6. Inhibition or knockdown of Igfbp5a, IGF1 receptor, PI3K, and Akt attenuates low [Ca2+]-induced ionocyte proliferation. The role of Trpv5/6 was investigated using a genetic mutant, targeted knockdown, and pharmacological inhibition. Loss-of-Trpv5/6 function or expression results in elevated pAkt levels and increased ionocyte proliferation under normal [Ca2+]. These increases are eliminated in the presence of an IGF1R inhibitor, suggesting that Trpv5/6 represses IGF1R-PI3K-Akt signaling under normal [Ca2+]. Intriguingly, blockade of Trpv5/6 activity inhibits the low [Ca2+]-induced activation of Akt. Mechanistic analyses reveal that the low [Ca2+]-induced IGF signaling is mediated through Trpv5/6-associated membrane depolarization. Low extracellular [Ca2+] results in a similar amplification of IGF-induced PI3K-PDK1-Akt signaling in human colon cancer cells in a TRPV6-dependent manner. These results uncover a novel and evolutionarily conserved signaling mechanism that contributes to the abnormal epithelial proliferation associated with Ca2+ deficiency.
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Abbreviations
- IGF:
-
insulin-like growth factors
- IGFBP:
-
insulin-like growth factor binding protein
- IGF1R:
-
insulin-like growth factor 1 receptor
- NaR cell:
-
Na+-K+-ATPase-rich cell
- TRPV:
-
transient receptor potential vanilloid
- CaM:
-
calmodulin
- hpf:
-
hour post fertilization
- HR cell:
-
H+-ATPase-rich cell
- NCC cell:
-
Na+/Cl− cotransporter cell
- pAkt:
-
phoshpo-Akt
- pErk:
-
phosphor-Erk
- CaSR:
-
Ca2+-sensing receptor
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Acknowledgements
We are grateful to Dr. Stefan Schulte-Merker, Hubrecht Institute, the Netherlands for providing the mus mutant fish line, Dr. Pung-Pung Hwang, Institute of Cellular and Organismal Biology, Academia Sinica in Taiwan, for sharing the double-label in situ hybridization protocol. We thank Dr. Haoxing Xu, University of Michigan, for reading and commenting on an earlier version of this manuscript. This work was supported by NIH Grant 1R21AG040604-01A1 and NSF Grant IOS-1051034 to CD.
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CD and WD designed the research; WD, YB, LH, XZ, JL, and J.K. performed research; CD, WD, YB, and XZ analyzed the data; and CD and WD wrote the paper.
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Dai, W., Bai, Y., Hebda, L. et al. Calcium deficiency-induced and TRP channel-regulated IGF1R-PI3K-Akt signaling regulates abnormal epithelial cell proliferation. Cell Death Differ 21, 568–581 (2014). https://doi.org/10.1038/cdd.2013.177
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DOI: https://doi.org/10.1038/cdd.2013.177
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