Expression of (NES-)hTERT in cancer cells delays cell cycle progression and increases sensitivity to genotoxic stress.
Telomerase is a reverse transcriptase associated with cellular immortality through telomere maintenance. This enzyme is activated in 90% of human cancers, and inhibitors of telomerase are currently in clinical trials to counteract tumor growth. Many aspects of telomerase biology have been investigated for therapy, particularly inhibition of the enzyme, but little was done regarding its subcellular shuttling. We have recently shown that mutations in the nuclear export signal of hTERT, the catalytic component of telomerase, led to a mutant ((NES-)hTERT) that failed to immortalize cells despite nuclear localization and catalytic activity. Expression of (NES-)hTERT in primary fibroblast resulted in telomere-based premature senescence and mitochondrial dysfunction. Here we show that expression of (NES-)hTERT in LNCaP, SQ20B and HeLa cells rapidly and significantly decreases their proliferation rate and ability to form colonies in soft agar while not interfering with endogenous telomerase activity. The cancer cells showed increased DNA damage at telomeric and extra-telomeric sites, and became sensitive to ionizing radiation and hydrogen peroxide exposures. Our data show that expression of (NES-)hTERT efficiently counteracts cancer cell growth in vitro in at least two different ways, and suggest manipulation with the NES of hTERT or its subcellular shuttling as a new strategy for cancer treatment.
Department of Pharmacology and Physiology, New Jersey Medical School, Newark, New Jersey, USA.
Olga A
OA
Kovalenko
Jessica
J
Kaplunov
Utz
U
Herbig
Sonia
S
Detoledo
Edouard I
EI
Azzam
Janine H
JH
Santos
D002448:Cell Adhesion; D002453:Cell Cycle; D045744:Cell Line, Tumor; D048430:Cell Shape; D004249:DNA Damage; D004272:DNA, Mitochondrial / Q000235:genetics; D006801:Humans; D008297:Male; D049790:Nuclear Export Signals / Q000235:genetics; D011471:Prostatic Neoplasms / Q000201:enzymology* / Q000473:pathology*; D012878:Skin Neoplasms / Q000201:enzymology* / Q000473:pathology*; D019098:Telomerase / Q000378:metabolism*; D016615:Telomere / Q000378:metabolism; D014410:Tumor Stem Cell Assay
D004272:DNA, Mitochondrial; D049790:Nuclear Export Signals; C509186:TERT protein, human; D019098:Telomerase