Genomic profiling of miRNAs in two human lens cell lines.
OBJECTIVE
Many miRNAs are expressed in a developmentally regulated and tissue-specific manner making them crucial for tissue development in a structure such as the eye. Since miRNA target function studies for the eye will need to be performed in ocular tissue culture cells, it is important to profile them for miRNA expression. Two commonly used human lens epithelial cell lines, HLE-B3 and SRA01/04, were profiled for miRNA.


METHODS
We performed miRNA profiling of two commonly used lens epithelial cell lines, HLE-B3 and SRA01/04. The differential expression levels detected for miR-184 and miR-31 were confirmed by qRT-PCR and the function of a predicted miR-184 target binding site was validated in-vitro.


RESULTS
We found that four miRNAs-miR-31, miR-124, miR-184, and miR-222-were differentially expressed between the two cell lines. We show that miR-184 binds to BIN3 3' UTR and while BIN3 mRNA expression was equal in both cell lines, the protein expression was inversely correlated with miR-184 expression.


CONCLUSIONS
The differences observed with respect to miRNA expression between two different lens epithelial cell lines were minimal. Still, caution will need to be exercised when choosing one cell line over another because of the expression differences for some miRNAs. Our results also suggest that miR-184 may regulate lens BIN3 expression in lens by a miRNA-mediated translational repression mechanism.
Tian
Lifeng
L
FM Kirby Center for Molecular Ophthalmology, Department of Ophthalmology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA. lifeng2@mail.med.upenn.edu
Huang
Kristen
K
DuHadaway
James B
JB
Prendergast
George C
GC
Stambolian
Dwight
D
D020413:3' Untranslated Regions / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D006801:Humans; D007908:Lens, Crystalline / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D020413:3' Untranslated Regions; D035683:MicroRNAs; D008840:Microfilament Proteins; D012333:RNA, Messenger; C430883:BIN3 protein, human; C509207:MIRN124 microRNA, human; C534424:MIRN184 microRNA, human; C525710:MIR222, human; C534612:MIRN31 microRNA, human