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  

Study on interference therapy induced by epidermal growth factor receptor-antisense cDNA in signal transduction of laryngeal squamous cell carcinoma

作　　者：鲜均明[1] 周光耀[1] 梁传余[1] 刘世喜[1] 

机构地区：[1]四川大学华西医院耳鼻咽喉科,四川成都610041

出　　处：《华西口腔医学杂志》2007年第6期540-543,547,共5页West China Journal of Stomatology

基　　金：国家自然科学基金资助项目(0040205401078);四川省青年科技基金资助项目(04ZQ026-024)

摘　　要：目的探讨表皮生长因子受体(EGFR)反义cDNA在体外对人喉癌Hep-2细胞信号转导的干预作用。方法以缺陷重组腺病毒为载体,构建EGFR正、反义cDNA的重组腺病毒,并在人胚肾母细胞(HEK293)中包装、纯化。将已纯化的EGFR cDNA重组腺病毒在体外转染人Hep-2喉癌细胞,采用MTT法检测重组腺病毒对Hep-2细胞的生长抑制作用、流式细胞术检测细胞周期DNA表达的变化、Western blot方法检测Hep-2细胞内EGFR蛋白的表达。结果成功地构建并制备了高滴度的EGFR cDNA 1 032 bp片段的正、反义重组腺病毒。同时,它们能被高效地转移到Hep-2细胞内,其中反义重组腺病毒能够抑制Hep-2细胞的增殖和EGFR蛋白的表达。结论EGFR反义cDNA能有效地干预人喉癌Hep-2细胞的细胞周期信号转导机制,从而抑制肿瘤细胞的增殖。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.

关 键 词：表皮生长因子受体 信号转导 喉癌 

分 类 号：R739.6[医药卫生—肿瘤]