EGFR反义cDNA联合p16β干预喉癌细胞信号转导的实验研究  

Interference Therapy of Laryngeal Squamous Cell Carcinoma by p16β and EGFR-Antisense cDNA in Signal Transduction

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,成都610041

出　　处：《四川大学学报（医学版）》2007年第4期569-572,共4页Journal of Sichuan University(Medical Sciences)

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

摘　　要：目的探讨表皮生长因子受体(EGFR)反义cDNA联合野生型p16β在体外对人喉癌Hep-2细胞信号转导的干预作用。方法将已纯化的EGFR反义cDNA重组腺病毒和p16β在体外转染Hep-2喉癌细胞,采用MTT实验、流式细胞术(FCM)、免疫组化、Westernblot等方法检测Hep-2细胞增殖抑制作用;进行细胞周期、DNA含量、细胞凋亡率的定量分析;检测重组腺病毒对Hep-2细胞EGFR蛋白表达的抑制和p16β蛋白过表达。结果反义EGFRmRNA表达重组腺病毒能有效抑制Hep-2细胞的增殖活性;超过77.7%以上的被转染Hep-2细胞被阻滞在G0/G1期;转染细胞的凋亡率升高;同时出现EGFR蛋白表达的抑制和p16β蛋白的过表达。当p16β重组腺病毒和EGFR反义cDNA重组腺病毒联合转染Hep-2细胞后其抑制Hep-2细胞增殖活性、细胞周期阻滞、抑制细胞调亡作用明细增强。结论p16β和EGFR反义cDNA能有效地干预人喉癌Hep-2细胞的信号转导机制,从而达到抑制Hep-2细胞增殖、诱导细胞凋亡的作用。Objective To test the effect of p16β and EGFR-antisense cDNA in signal transduction on Hep- 2 laryngeal squamous cell carcinoma in vitro. Methods The Hep-2 laryngeal squamous carcinoma cells were transfected by recombinant adenovirus AdEasy-EGFR-antisense and AdEasy-p16β in vitro. The inhibition of the EGFR expression and cell growth and changes of cell cycle, DNA content, apoptosis and ultramicrostructure of the Hep-2 cells were examined by MTT, Western blotting analysls, Flow cytometry analysis, Immunohistochemistry, and transmission electron microscope respectively. Results The proliferation of the Hep- 2 cells was inhibited significantly by the infection of the Ad- Ad-p16β or Ad-antisense EGFR. The infection also accelerated the apoptosis of the cancer cells. The proportion of cells in G0/G1 phases increased to more than 77.7%. The Ad-antisense EGFR-infected cells showed lower protein expression of EGFR. The P16β protein over expression was observed in the Ad-p16β-infected cells. Conclusion The transfection of Ad- Ad-p16β and Adantisense EGFR into Hep-2 cells leads to over-expression of Ad-p16β and under-expression of EGFR, along with Gl-phase arrest and apoptotic cell death. Both EGFR and Ad-p16β play important roles in the genesis, growth and differentiation of the human laryngocarcinoma cells.

关 键 词：表皮生长因子受体 细胞信号转导干预治疗 重组腺病毒 喉癌 p16β 

分 类 号：R739.65[医药卫生—肿瘤]