[Inhibition of hTR activity for suppression of lung cancer cell proliferation using antisense phosphorothioate oligonucleotides].
OBJECTIVE
Telomerase activity was measured in 8 lung tumor cell lines and transfected antisense phosphorothiate oligodeoxynucleotides (anti-PS-ODN) of hTR as therapeutic agents into target cell LTEP-a-2 to investigate the inhibitory effect on telomerase activity and tumor cell growth.


METHODS
1. Telomerase activity assay in 8 human lung tumor cell lines using telomerase PCR ELISA. 2. Synthesized anti-PS-ODNs of hTR and random PS-ODN transfected with Clonfectin into LTEP-a-2 cell lines for 72 hr. 3. Measure telomerase activity by telomerase PCR ELISA, SDH activity by MTT assay and cell growth.


RESULTS
Eight cell lines showed positive expression of telomerase activity. Various anti-PS-ODNs could inhibit telomerase activity, SDH activity and cell growth. The inhibition became more marked with the increase of anti-PS-ODNs concentrations. Concentrations of 5 - 40 micromol/L anti-PS-ODNs of hTR specifically reduced the telomerase activity by 25.7% - 84.0%. PS-ODNs of anti-hTR were also able to reduce SDH activity by 19.4% - 74.7% at 5 - 40 micromol/L. The dose of 5 - 40 micromol/L PS-ODNs of anti-hTR had the ability to inhibit cell growth by 14.8% - 72.5%. The results indicate that random sequence of PS-ODN(9) failed to inhibit telomerase, SDH activity and cell growth. A statistically significant difference exists between random PS-ODN and three anti-PS-ODNs (P < 0.01).


CONCLUSIONS
Human lung tumor cell lines express high telomerase activity. PS-ODNs of anti-hTR had the ability to inhibit telomerase and reduce LTEP-a-2 cell growth and metabolism.
Mei
M
M
Centre of molecular Biology, Tianjin Medical University, Tianjin 300070, China.
Yu
H
H
Zhang
W
W
Shi
J
J
Yang
J
J
D000230:Adenocarcinoma / Q000201:enzymology* / Q000473:pathology; D018286:Carcinoma, Giant Cell / Q000201:enzymology / Q000473:pathology; D002294:Carcinoma, Squamous Cell / Q000201:enzymology / Q000473:pathology; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D006801:Humans; D008175:Lung Neoplasms / Q000201:enzymology* / Q000473:pathology; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D012313:RNA / Q000235:genetics*; D013385:Succinate Dehydrogenase / Q000378:metabolism; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism*; D013873:Thionucleotides / Q000494:pharmacology
D020319:Oligodeoxyribonucleotides, Antisense; D013873:Thionucleotides; C422952:telomerase RNA; D012313:RNA; D013385:Succinate Dehydrogenase; D019098:Telomerase