ABSTRACT
<p><b>OBJECTIVE</b>To assess the telomerase activity inhibitory effect of ham merhead ribozyme targeting the 5'-end of human telomerase reverse transcriptase mRNA (hTERT-5'RZ), to compare it with the effect of another ribozyme teloRZ, and the combine the applications of the two ribozymes.</p><p><b>METHODS</b>hTERT-5'RZ gene was synthesized and cloned into pcDNA3.1(+); the ribozyme was produced by in vitro transcription. The teloRZ ribozyme was produced in the same way by in vitro transcription of p(SPT19-teloRZ) which had been constructed by the present authors. The ribozymes were transiently transfected into HeLa cells by liposome every 24 hours. After 72 hours, the cells were collected and their telomerase activities were assayed.</p><p><b>RESULTS</b>The ribozyme targeting the 5'-end of hTERT mRNA exhibited a very strong telomerase-inhibitory activity, the combined use of hTERT-5'RZ and teloRZ also showed clear inhibitory activity, but the inhibitory effect of teloRZ used alone was not so strong.</p><p><b>CONCLUSION</b>These observations suggest that the use of hTERT-5'RZ and the combined use of hTERT-5'RZ and teloRZ are more effective in telomerase inhibition as compare with the use of teloRZ alone. They may find applications in cancer therapy.</p>
Subject(s)
Humans , 5' Untranslated Regions , Cloning, Molecular , DNA-Binding Proteins , HeLa Cells , RNA, Catalytic , Metabolism , RNA, Messenger , Telomerase , Genetics , Transcription, GeneticABSTRACT
AIM: To observe the relationship between aortic atherosclerosis plaque formation and telomerase activation in vascular smooth muscle cells (VSMC). METHODS:Iliac artery atherosclerosis plaque from 20 renal failure patients and five cases of normal iliac arteries were obtained and confirmed by histological examination. The endothelial cells and the connective tissue on surface were lightly scraped and removed. Telomerase activity of the residual tissue was detected by telomeric repeat sequence amplification protocol. RESULTS:Five cases of normal iliac arteries showed negative telomerase activity. Eight of 20 cases athorosclerosis plaque showed positive telomerase activity, the other 12 cases showed negative telomerase activity. CONCLUSION: Telomerase activation may play a certain role in VSMC proliferation and atherosclerosis formation.