Determination of micro-RNA in cardiomyoblast cells using CE with LIF detection
- Determination of micro-RNA in cardiomyoblast cells using CE with LIF detection
- 반은미; 채동규; 송은주
- capillary electrophoresis; miRNA; Acute myocardial infarction; Biomarker; CE-LIF
- Issue Date
- VOL 34, NO 4, 598-604
- Micro-RNAs (miRNAs) are small, endogenous, singlestranded, and noncoding RNAs.
ThemiRNAs have been found to perform important functions in many cellular processes,
such as development, proliferation, differentiation, and apoptosis. Circulating miRNAs
have been proposed as emerging biomarkers in diseases such as cancer, diabetes, and
cardiovascular disease including acute myocardial infarction (AMI). In this study, we
developed CE with LIF (CE-LIF) using fluorescence-labeled DNA probe for determination
of low abundance miRNA in cell extracts. The target miRNA is miRNA-499, a biomarker
candidate of AMI with low abundance in biological samples. In order tomeasure the trace
level of miRNA, we optimized the hybridization conditions such as hybridization time,
temperature, and buffer solution. The highest fluorescence intensity of the hybridized
miRNA-499 was found when hybridization was conducted at 40℃ in 50 mM Tris-acetate
(pH 8.0) buffer containing 50 mM NaCl, and 10 mM EDTA for 15 min. The hybridized
miRNA-499 was detected in cultured H9c2 cardiomyoblast cells and the analysis ofmiRNA-
499 was completed within 1 h using CE-LIF. These results showed the potential of CE for
fast, specific, and sensitive high-throughput analysis of low-abundance miRNAs in cell
extracts, biofluids, and tissues.
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