RNAi抑制鼻咽癌细胞表皮生长因子受体表达对细胞生长的影响

Effect of silencing epidermal growth factor receptor expression by RNA interference on the growth of nasopharyngeal carcinoma cell 5-8F

目的利用RNAi技术抑制鼻咽癌细胞5-8F表皮生长因子受体(EGFR)表达,观察细胞生长变化,探讨EGFR与鼻咽癌发生发展之间的相关性。方法利用体外转录法合成EGFR特异性siRNA,将其瞬时转染5-8F,收集转染后细胞总RNA和蛋白,利用RT-PCR和Westernblot法测定EGFR表达水平的变化,并检测EGFR表达抑制后细胞生长速度和生长周期的变化。结果合成的3对siRNA中,有一对使EGFRmRNA表达下降67．5％,蛋白表达下降77％。EGFR表达抑制后5-8F生长速度减慢,并诱导细胞周期停止在G1期。结论RNAi沉默EGFR表达可抑制鼻咽癌细胞生长,并诱导癌细胞停止在G1期．RNAi可作为探索鼻咽癌发生发展机制和基因治疗的有效工具。

Objective To observe the growth and cell cycle changes ofnasopharyngeal carcinoma cell 5-8F in response to silencing of the expression of epidermal growth factor receptor （EGFR） with RNA interference （RNAi）, and explore the possible relationships between EGFR and the occurrence, differentiation and progression of nasopharyngeal carcinoma. Method Tl;lree EGFR-specific small interfering RNAs （siRNAs） were obtained by in vitro transcription and synthesis and were transiently transfected into 5-8F cells. The EGFR expression levels in the transfected cells were detected by reverse transcription （RT） -PCR and Western bloting, respectively. After EGFR expression silencing, the growth and cell cycle changes of the cells were observed. Results EGFR mRNA contents and protein levels decreased by approximately 67.5% and 77%, respectively, after RNAi of 5-8F cells, and the cell proliferation decreased and cell cycle arrest at GI phase occurred in association with EGFR silencing. Conclusion EGFR silencing by RNAi can reduce the proliferation ofnasopharyngeal carcinoma cells and induce cell cycle arrest at G1 phase, which sheds light on the possible use of RNAi for further investigation of the pathogenesis and gene theraov of nasooharvngeal carcinoma.