摘要
目的建立表达HPV16-E6/E7基因的人喉鳞癌细胞系。方法用阳离子脂质体基因转染技术将携带有HPV16-E6/E7DNA的真核表达质粒导入体外连续传20代的HPV阴性的Lscc-02喉鳞癌细胞系,采用PCR和Western blot检测HPV16-E6/E7基因在Lscc-02细胞系中的整合和表达。结果第22代和第35代细胞中,PCR和Western blot均分别检测出E6/E7基因和蛋白的存在,显示HPV16-E6/E7基因整合于Lscc-02细胞系中并能稳定表达。结论成功建立了表达HPV16-E6/E7基因的人喉鳞癌细胞系,为进一步研究HPV16-E6/E7基因在喉癌细胞中的作用机制以及喉癌的基因治疗奠定了基础。
Objective To establish a human laryngeal squamous cell carcinoma cell line expressing human papillomavirus （HPV16） E6/E7 oncogene. Methods Cationic phosphonolipid was used to transfect pLXSN16E6E7 into a HPV16-negative laryngeal squamous cell carcinoma cell line in passage number 20, Lsce-02. The expression of HPV16-F_.6/E7 in Lsce-02 cell line was detected by PCR and Western blot. Results HPV16-E6/E7 DNA was found in the transfeeted cell line by PCR and F_.6/E7 protein could be detected by Western blot in either passage number 22 or passage number 35, which showed that HPV16-E6/E7 was expressed stably in the transfeeted cell line. Conclusion A human laryngeal squamous cell carcinoma cell line expressing HPV16-E6/E7 has been established successfully. The newly established cell line may serve as a useful model for studying the role of HPV16-E6/E7 in laryngeal carcinoma cells and gene therapy of laryngeal carcinoma.
出处
《中国耳鼻咽喉颅底外科杂志》
CAS
2006年第6期418-421,共4页
Chinese Journal of Otorhinolaryngology-skull Base Surgery
