Efficient telomerase inhibition in human non-small cell lung cancer cells by liposomal delivery of 2'-O-methyl-RNA.
The antisense oligonucleotide 2'-O-methyl-RNA is a selective telomerase inhibitor targeting the telomerase RNA component and represents a potential candidate for anticancer therapy. The poor cellular uptake of 2'-O-methyl-RNA is a limiting factor that may contribute to the lack of functional efficacy. To improve delivery of 2'-O-methyl-RNA and consequently antitumoral efficiency in human lung cancer cells, we have investigated several transfection reagents. The transfection reagents DOTAP, MegaFectin 60, SuperFect, FuGENE 6 and MATra-A were tested for intracellular delivery. A FAM-labeled 2'-O-methyl-RNA was used to assess the intracellular distribution by confocal laser scanning microscopy in A549 human non-small cell lung cancer cells. Telomerase activity was measured using the telomeric repeat amplification protocol. Cell viability after transfection was quantified by the MTT assay. All transfection reagents enhanced 2'-O-methyl-RNA uptake in A549 cells but the cationic lipid reagents DOTAP and MegaFectin 60 were most efficient in the delivery of 2'-O-methyl-RNA resulting in telomerase inhibition. Among both DOTAP exhibited the lowest cytotoxicity. Our experiments show that DOTAP is the most suitable transfection reagent for the delivery of 2'-O-methyl-RNA in human lung cancer cells according to its relatively low cytotoxicity and its ability to promote efficient uptake leading to the inhibition of telomerase.
Beisner
Julia
J
Dr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology and University of Tübingen, Stuttgart, Germany.
Dong
Meng
M
Taetz
Sebastian
S
Piotrowska
Kamilla
K
Kleideiter
Elke
E
Friedel
Godehard
G
Schaefer
Ulrich
U
Lehr
Claus-Michael
CM
Klotz
Ulrich
U
Mürdter
Thomas E
TE
D000970:Antineoplastic Agents / Q000494:pharmacology*; D053801:Capillary Electrochromatography; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002784:Cholesterol / Q000737:chemistry / Q000633:toxicity; D016503:Drug Delivery Systems; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005079:Excipients; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry / Q000633:toxicity; D006801:Humans; D007202:Indicators and Reagents; D008081:Liposomes; D008175:Lung Neoplasms / Q000188:drug therapy*; D018613:Microscopy, Confocal; D009841:Oligonucleotides / Q000008:administration & dosage / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry / Q000633:toxicity; D016372:RNA, Antisense / Q000494:pharmacology*; D019098:Telomerase / Q000037:antagonists & inhibitors*; D013778:Tetrazolium Salts; D013844:Thiazoles; D014162:Transfection
D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D005079:Excipients; D005229:Fatty Acids, Monounsaturated; D007202:Indicators and Reagents; D008081:Liposomes; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D000644:Quaternary Ammonium Compounds; D016372:RNA, Antisense; D013778:Tetrazolium Salts; D013844:Thiazoles; D002784:Cholesterol; D019098:Telomerase; C022616:thiazolyl blue; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane