Synergistic down-regulation of telomerase by all-trans retinoic acid and antisense oligonucleotide in oral squamous cell carcinoma cell line (Tca8113).
Human telomerase, activated in about 90% of cancers, is mainly composed of hTR, hTERT and TP1. The exposed RNA template of hTR is an ideal target for antisense oligonucleotides (As-ODN); while recent findings indicate all-trans retinoid acid (ATRA) could effectively inhibit the expression of catalytic subunit-hTERT. The aim of this study was to investigate the effect of ATRA and As-ODN in oral squamous cell carcinoma and whether telomerase activity could be synergistically inhibited by them and thus therapeutically exploited in the future. As-ODN-hTR was transfected into human tongue squamous cell carcinoma cell line (Tca8113) with or without ATRA. Telomerase activity was examined by PCR-Elisa; viability was compared with growth curve; apoptotic rate was analyzed by Annexin V/PI double staining and hTERT expression was tested with western blot. Tca8113 cells displayed significant growth inhibition during the 9-day exposure to ATRA/As-ODN, especially to a combination of As-ODN-hTR and 5muM ATRA, correlating with the inhibition of telomerase expression. The relative telomerase activity was inhibited during treated with As-ODN-hTR alone, ATRA alone, or a combination of them. While without ATRA, the effect of As-ODN would disappear at 96h after transfection. As-ODN-hTR alone or combined with ATRA also significantly increase the apoptotic rate. Our findings provided direct evidence, in oral squamous cell carcinoma, As-ODN-hTR and ATRA could synergistically inhibit telomerase activity and telomerase protein in human tongue squamous cell carcinoma cells, which correlated with the induction of growth arrest.
Zhang
Ping
P
Department of Oral and Maxillofacial Surgery, The Ninth People's Hospital, Shanghai Second Medical University, 639 Zhizaoju Road, Shanghai 200011, China.
Xu
Qin
Q
Chen
Wan Tao
WT
Duan
Li Qiong
LQ
Zhang
Zhi Yuan
ZY
Zhou
Xiao Jian
XJ
D002294:Carcinoma, Squamous Cell / Q000201:enzymology* / Q000628:therapy; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors; D015536:Down-Regulation; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D019098:Telomerase / Q000037:antagonists & inhibitors*; D014062:Tongue Neoplasms / Q000201:enzymology* / Q000628:therapy; D014162:Transfection; D014212:Tretinoin / Q000494:pharmacology*
D004268:DNA-Binding Proteins; D016376:Oligonucleotides, Antisense; D014212:Tretinoin; D019098:Telomerase