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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.
Artigo em Chinês | WPRIM | ID: wpr-316256
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>
Assuntos
Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Patologia / Plasmídeos / Efeitos da Radiação / Proteínas Recombinantes / RNA Mensageiro / Ensaio de Imunoadsorção Enzimática / Carcinoma de Células Escamosas / Transfecção / Sobrevivência Celular / Neoplasias Laríngeas Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Oncology Ano de publicação: 2007 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Patologia / Plasmídeos / Efeitos da Radiação / Proteínas Recombinantes / RNA Mensageiro / Ensaio de Imunoadsorção Enzimática / Carcinoma de Células Escamosas / Transfecção / Sobrevivência Celular / Neoplasias Laríngeas Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Oncology Ano de publicação: 2007 Tipo de documento: Artigo