表皮生长因子受体反义cDNA干预喉癌细胞信号转导的实验研究

Objective To investigate effect of interference therapy induced by epidermal growth factor receptor （ EGFR）- antisense cDNA in signal transduction of Hep - 2 laryngeal squamous cell carcinoma in vitro. Methods AdEasy Vector System was used to construct the recombinant adenovirus vector sense/antisense- pAdEasy- EGFR. The recombinant adenovirus vector introduced EGFR- sense/antisense cDNA fragment into HEK293 cell. The purified recombinant adenovirus sense/antisense- pAdEasy- EGFR transfected Hep- 2 cells in vitro. The inhibition of EGFR protein expression and proliferation of Hep- 2 cells, the changes of cell cycle and DNA content in Hep- 2 cells were examined by MTT, Western blot analysis, flow cytometry essay, and immunocytochemistry respectively. Results The higher titre sense and antisense mRNA expression recombinant adenovirus containing 1 032 bp EGFR- cDNA was constructed and prepared successfully. When antisense- pAdEasy- EGFR was transferred into Hep- 2 cells the inhibition of cell proliferation and EGFR protein expression in Hep- 2 cells were investigated effectively. Conclusion The antisense- pAdEasy- EGFR effectively interfere the Hep- 2 signal transduction pathway and induce apoptosis which results in inhibiting proliferation of laryngeal squamous cell carcinoma.