Simultaneous detection of multiple microRNAs for expression profiles of microRNAs in lung cancer cell lines by capillary electrophoresis with dual laser-induced fluorescence

• Development of CE with dual LIF detection method for the simultaneous detection of multiple miRNAs.
• Determination and partial validation of three miRNAs in cells using CE with dual LIF detection system.
• Simultaneous screening of three miRNAs in various lung cancer cell lines using CE with dual LIF detection system.

MicroRNAs (miRNAs) are small, endogenous, single-stranded, noncoding RNAs. Circulating miRNAs are being considered as promising disease biomarkers. Indeed, single miRNAs have been associated with a wide variety of disease conditions and can target multiple mRNAs; therefore, several miRNAs may be simultaneously involved in disease progression and development. In this study, we developed a capillary electrophoresis with dual laser-induced fluorescence (CE with dual LIF) method using two color laser excitations for simultaneous determination of multiple miRNAs. Target miRNAs were hybridized with 6-FAM- or Cy5-labeled DNA probes for simultaneous determination of multiple miRNAs at excitation wavelengths of 488 and 635 nm. The hybridized miRNAs were separated using CE with dual LIF and detected within 13 min at excitation wavelengths of 488 and 635 nm without any interference or crosstalk. Additionally, the proposed approach was used successfully to detect and evaluate levels of several endogenous miRNAs from lung cancer cell lines. These results showed the potential of CE with dual LIF for fast, specific, simultaneous analysis of multiple miRNAs in cell extracts, biofluids, and tissues.