ERalpha increases expression and interacts with TERT in cataractous canine lens epithelial cells.
OBJECTIVE
Estrogen receptor alpha (ERalpha) expression has previously been evaluated in lens epithelial cells (LEC). However, its function in the lens has not been determined. One potential function may be its interaction with the catalytic subunit of telomerase (TERT), which is present in normal LEC and higher in LEC that have undergone epithelial to mesenchymal transition (EMT). ERalpha is known to play a role in EMT, a process that may also involve TERT.


METHODS
A commercially available transcription factor array was used to evaluate potential interactions between TERT and other proteins in normal and cataractous LEC. Based on these findings, ERalpha protein and mRNA expressions were measured using western blot analysis, immunohistochemical staining, and quantitative reverse transcription polymerase chain reaction (RT-PCR). Co-immunoprecipitation assays were used to evaluate the interaction of TERT with ERalpha as well as their phosphorylation in normal and cataractous LEC.


RESULTS
The transcription factor array suggested that TERT interacted with ERalpha via the estrogen response element (ERE) in cataractous LEC but not in normal LEC. Expression of ERalpha protein and mRNA increased in cataractous LEC compared with normal LEC. Co-immunoprecipitation assays confirmed the interaction of TERT with ERalpha in cataractous LEC while no interaction was found in normal LEC. LEC that have undergone EMT, e.g., cataracts, are rapidly proliferating and migrating along the posterior lens capsule.


CONCLUSIONS
ERalpha is known to play a role in EMT, and our data suggests that TERT and phosphorylated protein kinase B (pAkt) may be involved in the regulation of this process in cataractous LEC.
Colitz
Carmen M H
CM
Veterinary Clinical Sciences, Ohio State University, Columbus, OH, USA. ccolitz@gmail.com
Sugimoto
Yasuro
Y
Lu
Ping
P
Barden
Curtis Andrew
CA
Thomas-Ahner
Jennifer
J
Chandler
Heather Lynn
HL
D000818:Animals; D015153:Blotting, Western; D002386:Cataract / Q000201:enzymology* / Q000235:genetics / Q000473:pathology*; D002457:Cell Extracts; D004285:Dogs; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology*; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism*; D047629:Estrogen Receptor beta / Q000378:metabolism; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D007908:Lens, Crystalline / Q000378:metabolism / Q000473:pathology*; D010766:Phosphorylation; D011485:Protein Binding; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase / Q000378:metabolism*
D002457:Cell Extracts; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D019098:Telomerase