Published online 28 September 2005
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Real-time expression profiling of microRNA precursors in human cancer cell lines
1College of Pharmacy, Ohio State University Columbus, OH, USA 2Department of Surgery, University of Oklahoma Health Sciences Center Oklahoma City, OK, USA
*To whom correspondence should be addressed. Tel: +1 614 292 3456; Fax: +1 614 292 7766; Email: schmittgen.2{at}osu.edu
Received April 27, 2005. Revised August 23, 2005. Accepted September 7, 2005.
Our previous study described a real-time PCR method to quantify microRNA (miRNA) precursors using SYBR green detection [T. D. Schmittgen, J. Jiang, Q. Liu and L. Yang (2004) Nucleic Acids Res., 32, e43]. The present study adapted the assay to a 384-well format and expanded it to include primers to 222 human miRNA precursors. TaqMan minor groove binder probes were used to discriminate nearly identical members of the let-7 family of miRNA isoforms. The miRNA precursor expression was profiled in 32 human cell lines from lung, breast, colorectal, hematologic, prostate, pancreatic, and head and neck cancers. Some miRNA precursors were expressed at similar levels in many of the cell lines, while others were differentially expressed. Clustering analysis of the miRNA precursor expression data revealed that most of the cell lines clustered into their respective tissues from which each cell line was ostensibly derived. miRNA precursor expression by PCR paralleled the mature miRNA expression by northern blotting for most of the conditions studied. Our study provides PCR primer sequences to all of the known human miRNA precursors as of December 2004 and provides a database of the miRNA precursor expression in many commonly used human cancer cell lines.
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