Z Shen1, D Ye, X Zhang, Z Jiang, B Xiao, J Guo. 1. Department of Otorhinolaryngology-Head and Neck Surgery, Lihuili Hospital of Ningbo University, China. szs7216@sina.com
Abstract
OBJECTIVES: To investigate the effect of the HuR gene on laryngeal carcinoma Hep-2 cell growth, and to analyse correlations between the HuR, cyclooxygenase-2 and survivin genes. STUDY DESIGN: Experiment study. SETTING: Department of Otolaryngology-Head and Neck Surgery, Lihuili Hospital of Ningbo University, Ningbo, a tertiary care centre in China. METHODS: Copies of a small interfering RNA segment directed against the HuR gene were transfected into Hep-2 cells using Lipofectamine™ 2000. The effect of the small interfering RNA segment on Hep-2 cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Changes in the expression of the HuR, cyclooxygenase-2 and survivin genes were detected by semi-quantitative reverse transcription polymerase chain reaction analysis. Concentrations of the HuR, cyclooxygenase-2 and survivin proteins were evaluated using Western blotting. RESULTS: Expression of the HuR, cyclooxygenase-2 and survivin genes, as indicated by messenger RNA and protein levels, was suppressed by the HuR gene small interfering RNA segment in a dose-dependent manner. The proliferation indices of all treated groups were significantly lower than those of control groups (p < 0.05). CONCLUSIONS: Impairment of HuR gene expression, using interfering RNA technology, can significantly suppress Hep-2 cell proliferation and induce apoptosis. The HuR gene may be an effective target for gene therapy in patients with laryngeal carcinoma.
OBJECTIVES: To investigate the effect of the HuR gene on laryngeal carcinoma Hep-2 cell growth, and to analyse correlations between the HuR, cyclooxygenase-2 and survivin genes. STUDY DESIGN: Experiment study. SETTING: Department of Otolaryngology-Head and Neck Surgery, Lihuili Hospital of Ningbo University, Ningbo, a tertiary care centre in China. METHODS: Copies of a small interfering RNA segment directed against the HuR gene were transfected into Hep-2 cells using Lipofectamine™ 2000. The effect of the small interfering RNA segment on Hep-2 cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Changes in the expression of the HuR, cyclooxygenase-2 and survivin genes were detected by semi-quantitative reverse transcription polymerase chain reaction analysis. Concentrations of the HuR, cyclooxygenase-2 and survivin proteins were evaluated using Western blotting. RESULTS: Expression of the HuR, cyclooxygenase-2 and survivin genes, as indicated by messenger RNA and protein levels, was suppressed by the HuR gene small interfering RNA segment in a dose-dependent manner. The proliferation indices of all treated groups were significantly lower than those of control groups (p < 0.05). CONCLUSIONS: Impairment of HuR gene expression, using interfering RNA technology, can significantly suppress Hep-2 cell proliferation and induce apoptosis. The HuR gene may be an effective target for gene therapy in patients with laryngeal carcinoma.