Detection of a long non-coding RNA (CCAT1) in living cells and human adenocarcinoma of colon tissues using FIT-PNA molecular beacons

Yossi Kam, Abraham Rubinstein, Shankar Naik, Irena Djavsarov, David Halle, Ilana Ariel, Ali O. Gure, Alexander Stojadinovic, Hong Guang Pan, Victoria Tsivin, Aviram Nissan*, Eylon Yavin

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

97 Scopus citations

Abstract

Although the function and mechanism of action of long non-coding RNAs (lncRNA) is still not completely known, studies have shown their potential role in the control of gene expression and regulation, in cellular proliferation and invasiveness at the transcriptional level via multiple mechanisms. Recently, colon cancer associated transcript 1 (CCAT1) lncRNA was found to be expressed in colorectal cancer (CRC) tumors but not in normal tissue. This study aimed to study the ability of a CCAT1-specific peptide nucleic acid (PNA) based molecular beacons (TO-PNA-MB) to serve as a diagnostic probe for in vitro, ex vivo, and in situ (human colon biopsies) detection of CRC. The data showed enhanced fluorescence upon in vitro hybridization to RNA extracted from CCAT1 expressing cells (HT-29, SW-480) compared to control cells (SK-Mel-2). Uptake of TO-PNA-MBs into cells was achieved by covalently attaching cell penetrating peptides (CPPs) to the TO-PNA-MB probes. In situ hybridization of selected TO-PNA-MB in human CRC specimens was shown to detect CCAT1 expression in all (4/4) subjects with pre-cancerous adenomas, and in all (8/8) patients with invasive adenocarcinoma (penetrating the bowel wall) tumors. The results showed that CCAT1 TO-PNA-MB is a powerful diagnostic tool for the specific identification of CRC, suggesting that with the aid of an appropriate pharmaceutical vehicle, real time in vivo imaging is feasible. TO-PNA-MB may enable identifying occult metastatic disease during surgery, or differentiating in real time in vivo imaging, between benign and malignant lesions.

Original languageAmerican English
Pages (from-to)90-96
Number of pages7
JournalCancer Letters
Volume352
Issue number1
DOIs
StatePublished - 28 Sep 2014

Bibliographical note

Funding Information:
The results reported here are included in the dissertation Project of Y.K. in partial fulfillment of his Ph.D. degree requirements at The Hebrew University of Jerusalem. E.Y. acknowledges the David R. Bloom Center for Pharmacy and the Grass Center for Drug Design and Synthesis of Novel Therapeutics for financial support. A.R. acknowledges the Israel Cancer Association for financial support.

Keywords

  • Colorectal cancer
  • Imaging
  • Long non-coding RNA
  • Molecular beacon
  • Peptide nucleic acid

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