Effective inhibition of hTERT expression by RNA interference on the radiosensitivity of human laryngeal cancer Hep-2 cell line / 中华肿瘤杂志
Chinese Journal of Oncology
; (12): 9-13, 2007.
Article
Dans Zh
| WPRIM
| ID: wpr-316256
Responsable en Bibliothèque :
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct an eukaryotic expression vector of human telomerase reverse transcriptase (hTERT) gene specific shRNA, and investigate the effect of pshRNA-hTERT combined with gamma-irradiation on cell survival and telomerase activity.</p><p><b>METHODS</b>According to the coding sequence of hTERT mRNA, the target of RNAi was designed, and recombinant expression plasmid pshRNA-hTERT was constructed. The vector was transfected into Hep-2 cells. The radiosensitivity of Hep-2 cells was determined by clonogenic assay. Telomeric repeat amplification protocol (TRAP-PCR-ELISA) was used to observe the telomerase activity in each group. Results Recombinant expression vector pshRNA-hTERT was successfully transfected into Hep-2 cells. The hTERT expression inhibition rate reached 60. 8%. pshRNA-hTERT not only inhibited telomerase activity of Hep-2, but also inhibited the raise of telomerase activity induced by gamma-irradiation. Exposure of Hep-2 cells to pshRNA-hTERT for 24 hrs before irradiation resulted in a decrease in mean surviving fraction of Hep-2 cells compared with cells of group with irradiation alone (67. 7% vs 85. 7%, P <0. 05) .</p><p><b>CONCLUSION</b>RNAi showed a significant inhibitory effect to the expression of hTERT. The results indicate that pshRNA-hTERT can effectively inhibit telomerase activity of Hep-2 cells treated or untreated with 2 Gy gamma-irradiation and significantly enhance the radiosensitivity of Hep-2 cells in vitro. The role of radiosensitization of pshRNA-hTERT may be related with the inhibition of telomerase activity.</p>
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Anatomopathologie
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Plasmides
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Effets des rayonnements
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Protéines recombinantes
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ARN messager
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Test ELISA
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Carcinome épidermoïde
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Transfection
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Survie cellulaire
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Tumeurs du larynx
Limites du sujet:
Humans
langue:
Zh
Texte intégral:
Chinese Journal of Oncology
Année:
2007
Type:
Article