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- W3023408049 abstract "MicroRNA (miRNA) is a recently identified paradigm of gene regulation that involves small RNA of approximately 22 nucleotides. miRNA have been described as “biological dark matter”, are evolutionarily conserved from plants to animals, C. elegans to humans, and have been found to be critical to development and carcinogenesis. However, we are not aware of previous reports on whether miRNA levels are affected by radiation in human cells. To assess the levels of a panel of miRNAs (miR-1, miR-17–5p, miR-21, miR-29, miR-133, Let-7c, Let-7g) in human cancer cells (HCT 116, HeLa, PC3, SKBr3) in response to irradiation (ionizing and ultraviolet (UV)). A panel of human cancer cells was mock-treated, or treated with ionizing (IR) or ultraviolet (UV) radiation. Total RNA was extracted from each treatment group and then enriched for small RNAs using a filter-based isolation system (mirVANA (Ambion)). The amount of RNA was standardized to cell number, confirmed via spectrophotometry, and normalized to levels of tRNA and U6 small nuclear RNA. The extracted RNA was separated on denaturing polyacrylamide-urea gels via electrophoresis and then transferred to nylon membranes. The membranes were then probed with radiolabeled probes specific for each miRNA, followed by autoradiography. Cell-type specific expression patterns of miRNA were readily detected. In Hela cells, miR-21 and miR-17–5p were increased after IR, while Let-7g was decreased (see Figure; tRNA served as a loading control gel). Both miR-21 and miR-17–5p were also increased after IR in HCT116, but not in SKBR3 and PC3 cells. Finally, Let-7g was not decreased after IR in HCT116, SKBR3, or PC3. Surprisingly, UV did not substantially alter the levels of most of the miRNAs. The functional effects of downregulating miR-21 will be presented. Cell-type specific patterns of miRNA expression are readily detectable in human cancer cells. Levels of specific miRNAs are changed after IR, but this response is also cell-type specific. Finally, the pattern of miRNA expression after IR can be distinguished from UV. Together, these results suggest an unexpected complexity in the response of miRNAs after irradiation. The increase in specific miRNAs after IR may be consistent with pro-survival functions, as suggested by previous reports." @default.
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- W3023408049 date "2006-11-01" @default.
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- W3023408049 title "2632" @default.
- W3023408049 doi "https://doi.org/10.1016/j.ijrobp.2006.07.1046" @default.
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