Stathmin表达抑制的鼻咽癌细胞系建立

目的 建立stathmin表达抑制的鼻咽癌细胞系,为stathmin在鼻咽癌中的生物学功能研究提供实验基础.方法 合成stathmin特异性干扰DNA片段,将合成的干扰DNA片段克隆到siRNA质粒表达载体pGenesil-1.1,载体导入JM109菌株进行扩增,酶切和测序对重组表达载体进行鉴定.应用脂质体将质粒表达载体转入鼻咽癌5-8F细胞,采用G418进行筛选；RT-PCR和Western blot检测stathmin表达.结果 酶切和测序分析证实,插入siRNA质粒表达载体中的stathmin干扰DNA片段的序列和方向正确.表达分析表明,构建的stathmin特异性siRNA质粒表达载体能有效沉默stathmin在鼻咽癌5-8F细胞中的表达.结论 sathmin表达抑制的鼻咽癌细胞系建立成功.

Construction of stathmin knock-down nasopharyngeal cancer cell line

【Objective】 To establish a stable stathmin knock-down nasopharyngeal carcinoma cell line for providing experimental base to the biological function research of stathmin in nasopharyngeal carcinoma.【Methods】 Synthesizing stathmin-specific interfering DNA fragment and subcloning the fragment into siRNA expression plasmid vector pGenesil-1.1 which was imported JM109 for amplification.The subcloning vectors were identified by enzyme digestion and sequence analysis.The identified vector was transfected nasopharyngeal carcinoma 5-8F cell line by liposome.After G418 selection,stathmin expression was analyzed by RT-PCR and western blot.【Results】 The subcloning vector had gotten stathmin interfering DNA fragment with correct direction.Expression analysis showed that the built plasmid vector could effectively knock down stathmin expression in nasopharyngeal carcinoma 5-8F cell line.【Conclusion】 Stathmin expression suppressing nasopharyngeal carcinoma cell line has been successfully constructed.