TY - JOUR
T1 - Simultaneous detection of multiple microRNAs for expression profiles of microRNAs in lung cancer cell lines by capillary electrophoresis with dual laser-induced fluorescence
AU - Ban, Eunmi
AU - Chae, Dong Kyu
AU - Song, Eun Joo
N1 - Funding Information:
We gratefully acknowledge the support of the GRL Program ( 20110021713 ) from the National Research Foundation of Korea and an institutional grant from Korea Institute of Science and Technology ( 2E23810 and 2E24080 ).
PY - 2013/11/8
Y1 - 2013/11/8
N2 - 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.
AB - 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.
KW - Biomarker
KW - CE with dual LIF
KW - Clinical diagnosis
KW - Lung cancer
KW - MiRNA
KW - Multiple detection
UR - http://www.scopus.com/inward/record.url?scp=84885382238&partnerID=8YFLogxK
U2 - 10.1016/j.chroma.2013.09.048
DO - 10.1016/j.chroma.2013.09.048
M3 - Article
C2 - 24094482
AN - SCOPUS:84885382238
SN - 0021-9673
VL - 1315
SP - 195
EP - 199
JO - Journal of Chromatography A
JF - Journal of Chromatography A
ER -