title: Detection of a long non-coding RNA (CCAT1) in living cells and human adenocarcinoma of colon tissues using FIT-PNA molecular beacons.
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.
authors: The Hebrew University of Jerusalem, Faculty of Medicine, The School of Pharmacy, Institute for Drug Research, P.O. Box 12065, Jerusalem 91120, Israel.
authors: Yossi
authors: Y
authors: Kam
authors: The Hebrew University of Jerusalem, Faculty of Medicine, The School of Pharmacy, Institute for Drug Research, P.O. Box 12065, Jerusalem 91120, Israel; The Harvey M. Krueger Family Center for Nanoscience and Nanotechnology and the David R. Bloom Center of Pharmacy of the Hebrew University of Jerusalem, Israel.
authors: Abraham
authors: A
authors: Rubinstein
authors: The Hebrew University of Jerusalem, Faculty of Medicine, The School of Pharmacy, Institute for Drug Research, P.O. Box 12065, Jerusalem 91120, Israel.
authors: Shankar
authors: S
authors: Naik
authors: The Hebrew University of Jerusalem, Faculty of Medicine, The School of Pharmacy, Institute for Drug Research, P.O. Box 12065, Jerusalem 91120, Israel.
authors: Irena
authors: I
authors: Djavsarov
authors: Department of Surgery, Hadassah-Hebrew University Medical Center, Ein Kerem, P.O. Box 12000, Jerusalem 91120, Israel.
authors: David
authors: D
authors: Halle
authors: Department of Pathology, Hadassah-Hebrew University Medical Center, Mount Scopus, P.O. Box 24035, Jerusalem 91240, Israel.
authors: Ilana
authors: I
authors: Ariel
authors: Department of Molecular Biology and Genetics, SB-248 Bilkent University Ankara, Turkey.
authors: Ali O
authors: AO
authors: Gure
authors: Department of Surgery, Walter Reed National Military Medical Center, Bethesda, MD, USA.
authors: Alexander
authors: A
authors: Stojadinovic
authors: Department of Surgery, Walter Reed National Military Medical Center, Bethesda, MD, USA.
authors: HongGuang
authors: H
authors: Pan
authors: Department of Surgery, Hadassah-Hebrew University Medical Center, Ein Kerem, P.O. Box 12000, Jerusalem 91120, Israel.
authors: Victoria
authors: V
authors: Tsivin
authors: Department of Surgery, Hadassah-Hebrew University Medical Center, Ein Kerem, P.O. Box 12000, Jerusalem 91120, Israel. Electronic address: anissan@hadassah.org.il.
authors: Aviram
authors: A
authors: Nissan
authors: The Hebrew University of Jerusalem, Faculty of Medicine, The School of Pharmacy, Institute for Drug Research, P.O. Box 12065, Jerusalem 91120, Israel. Electronic address: eylony@ekmd.huji.ac.il.
authors: Eylon
authors: E
authors: Yavin
mesh_terms: D000230:Adenocarcinoma / Q000175:diagnosis / Q000473:pathology* / Q000503:physiopathology; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000175:diagnosis / Q000473:pathology* / Q000503:physiopathology; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D018613:Microscopy, Confocal; D020135:Peptide Nucleic Acids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D062085:RNA, Long Noncoding / Q000097:blood / Q000235:genetics / Q000302:isolation & purification*
chemical_list: C579842:CCAT1 long noncoding RNA, human; D020135:Peptide Nucleic Acids; D062085:RNA, Long Noncoding
keywords: Colorectal cancer; Imaging; Long non-coding RNA; Molecular beacon; Peptide nucleic acid