Mature microRNAs identified in highly purified nuclei from HCT116 colon cancer cells

Chang Won Park, Yan Zeng, Xiaoxiao Zhang, Subbaya Subramanian, Clifford J. Steer

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

MicroRNAs (miRNAs) have emerged as one of the major regulatory mechanisms of gene expression. A major function of miRNAs involves the post-transcriptional regulation of target mRNAs, which is reported to occur primarily in the cytoplasm. However, there is a significant amount of evidence demonstrating the existence of small non-coding RNAs, including small-interfering RNA (siRNA), miRNA and Piwi-interacting RNA (piRNA) in the nucleus. In order to elucidate the potential subcellular localizations and functions of miRNAs, we have identified numerous miRNAs that are present in isolated nuclei from human colon cancer HC T116 cells. MicroRNA profiles were compared between cytoplasmic and nuclear fractions of the HC T116 cell line on the basis of multiple microarray analyses. MicroRNA species showing significant existence in isolated and highly purified populations of nuclei were selected and further tested with RT-PC R. The nuclear localization of the mature form of miRNAs was verified again by control RT-PC R excluding the detection of premature forms of miRNA, such as pri-miRNA or pre-miRNA. The elevated levels of representative miRNAs identified in purified nuclei were confirmed by northern blot analysis, supporting the notion that significant numbers of mature miRNAs exist not only in the cytoplasm but also in the nucleus. These results will likely provide a basis for further studies concerning the intracellular trafficking and nuclear location of miRNAs.

Original languageEnglish (US)
Pages (from-to)606-614
Number of pages9
JournalRNA Biology
Volume7
Issue number5
DOIs
StatePublished - 2010

Bibliographical note

Funding Information:
The authors would like to thank Thomas J. Bedor for technical assistance. This research was supported, in part, by ARRA NIH Grant R01 DK081865-01 (to C. J. Steer).

Keywords

  • Cytoplasmic
  • Microarray
  • Northern blot
  • Nucleus
  • RT-PCR
  • microRNA

Fingerprint Dive into the research topics of 'Mature microRNAs identified in highly purified nuclei from HCT116 colon cancer cells'. Together they form a unique fingerprint.

Cite this