Abstract
Purpose::
To comprehensively investigate the MicroRNAs (miRNAs) profile among retinoblastoma cell lines, retinoblastoma tissues and human fetal retinal tissues.
Methods::
We used approximately 105 cells from retinoblastoma cell lines or 2-10mg of tissues from retinoblastoma or human fetal retinal layer to isolate the low molecular weight RNA using the Ambion RNA kit. Real-time PCR was performed with the TaqMan MicroRNA Assays Human Panel protocol on an Applied Biosystems 7900HT Sequence Detection System.
Results::
We studied the miRNA profiles of 2 retinoblastoma cell lines (Y79, WERI-Rb1), 2 microdissected retinoblastoma tissues and 2 fetal retinae at gestational week 12 using the TaqMan MicroRNA Assays. Among the 157 miRNAs studied, we identified 5 miRNAs that were aberrantly up-regulated (4-60 fold) in retinoblastoma tissues and cell lines when normalized with hsa-let7a. In the retinoblastoma tissue, we also recognized the presence and up-regulation of miRNAs that there were well characterized to be associated with cancer, such as hsa-miR-93. Comparing with other cancer cell lines (Hela and MCF-7), we identified hsa-miR-181 and hsa-miR-183 to be specifically up-regulated in retinal tissues. These two miRNAs had been reported to be highly expressed in mouse retina.
Conclusions::
Our findings support the notion that miRNAs of the human eye display distinctive and overlapping tissue specificity. In addition, the differentially up or down regulated miRNAs identified in retinoblastoma tissue may contribute to its tumorigenesis.
Keywords: retinoblastoma • retina