Cyclic induction of senescence with intermittent AZT treatment accelerates both apoptosis and telomere loss.
BACKGROUND
3'-azido-2',3'-dideoxythymidine (AZT) is phosphorylated intracellularly to 3'-azido-3'-deoxythymidine-5'-triphosphate (AZT-TP), which is incorporated into telomeric DNA, thereby blocking chain elongation. AZT is also known to inhibit reverse transcriptase, as well as other cellular enzymes including DNA polymerase gamma, thymidine kinase, and telomerase.


METHODS
We induced cancer cell senescence by treating MCF-7 cells with AZT in dosages of IC10 and IC20 for an extended period (about 120 population doublings (PD)). We then investigated the sequential changes in cellular growth, expression of telomerase subunits and transcription factors (c-Myc, Mad1), telomerase activity and telomere length.


RESULTS
Senescence, apoptosis, growth delay, inhibition of telomerase activity and shortening of telomere length were all observed in a dose- and time-dependent manner. After the onset of senescence, the apoptosis rate increased slowly during early PDs. In contrast to senescence, the apoptotic rate showed little change after AZT removal, while it increased suddenly and significantly in a dose-dependent manner upon the second introduction of AZT. Continuous shortening of the telomeric length was observed with AZT, and, upon re-exposure to AZT, shortening of the telomere occurred more rapidly than with first exposure. Of the telomerase subunits, telomerase reverse transcriptase (hTERT) and c-Myc were the first to show a reduction in activity after AZT treatment, followed by changes in hTER , Mad1 and hTEP-1.


CONCLUSIONS
Cyclic treatment with AZT initially suppressed hTERT and c-Myc, followed by suppression of hTER, Mad1 and hTEP-1. Furthermore, the treatment accelerated both telomere loss and apoptosis, even when administered at a senescence-inducing dosage level.
Ji
Hyun Jung
HJ
Cancer Metastasis Research Center, Young Cancer Center, Yonsei University College of Medicine, Seoul, Seodaemun-Ku, Korea.
Rha
Sun Young
SY
Jeung
Hei Cheul
HC
Yang
Sang Hwa
SH
An
Sung Whan
SW
Chung
Hyun Cheol
HC
D017209:Apoptosis / Q000187:drug effects*; D001943:Breast Neoplasms / Q000188:drug therapy*; D016922:Cellular Senescence / Q000187:drug effects / Q000235:genetics; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology*; D019098:Telomerase / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D016615:Telomere / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D014157:Transcription Factors / Q000187:drug effects / Q000378:metabolism; D014407:Tumor Cells, Cultured; D015215:Zidovudine / Q000494:pharmacology*
D004268:DNA-Binding Proteins; C116115:MYCBP protein, human; D018894:Reverse Transcriptase Inhibitors; D014157:Transcription Factors; D015215:Zidovudine; D019098:Telomerase